1909328
Rapid Effects of Hearing Song on Catecholaminergic Activity in the Songbird Auditory Pathway
Catecholaminergic (CA) neurons innervate sensory areas and affect the processing of sensory signals. For example, in birds, CA fibers innervate the auditory pathway at each level, including the midbrain, thalamus, and forebrain. We have shown previously that in female European starlings, CA activity in the auditory forebrain can be enhanced by exposure to attractive male song for one week. It is not known, however, whether hearing song can initiate that activity more rapidly. Here, we exposed estrogen-primed, female white-throated sparrows to conspecific male song and looked for evidence of rapid synthesis of catecholamines in auditory areas. In one hemisphere of the brain, we used immunohistochemistry to detect the phosphorylation of tyrosine hydroxylase (TH), a rate-limiting enzyme in the CA synthetic pathway. We found that immunoreactivity for TH phosphorylated at serine 40 increased dramatically in the auditory forebrain, but not the auditory thalamus and midbrain, after 15 min of song exposure. In the other hemisphere, we used high pressure liquid chromatography to measure catecholamines and their metabolites. We found that two dopamine metabolites, dihydroxyphenylacetic acid and homovanillic acid, increased in the auditory forebrain but not the auditory midbrain after 30 min of exposure to conspecific song. Our results are consistent with the hypothesis that exposure to a behaviorally relevant auditory stimulus rapidly induces CA activity, which may play a role in auditory responses.
535645
An integrative theory of locus coeruleus-norepinephrine function: adaptive gain and optimal performance.
Historically, the locus coeruleus-norepinephrine (LC-NE) system has been implicated in arousal, but recent findings suggest that this system plays a more complex and specific role in the control of behavior than investigators previously thought. We review neurophysiological and modeling studies in monkey that support a new theory of LC-NE function. LC neurons exhibit two modes of activity, phasic and tonic. Phasic LC activation is driven by the outcome of task-related decision processes and is proposed to facilitate ensuing behaviors and to help optimize task performance (exploitation). When utility in the task wanes, LC neurons exhibit a tonic activity mode, associated with disengagement from the current task and a search for alternative behaviors (exploration). Monkey LC receives prominent, direct inputs from the anterior cingulate (ACC) and orbitofrontal cortices (OFC), both of which are thought to monitor task-related utility. We propose that these frontal areas produce the above patterns of LC activity to optimize utility on both short and long timescales.
10004500
A matter of focus: monoaminergic modulation of stimulus coding in mammalian sensory networks
Although the presence of neuromodulators in mammalian sensory systems has been noted for some time, a groundswell of evidence has now begun to document the scope of these regulatory mechanisms in several sensory systems, highlighting the importance of neuromodulation in shaping feature extraction at all levels of neural processing. The emergence of more sophisticated models of sensory encoding and of the interaction between sensory and regulatory regions of the brain will challenge sensory neurobiologists to further incorporate a concept of sensory network function that is contingent on neuromodulatory and behavioral state.
4353142
Cortical remodelling induced by activity of ventral tegmental dopamine neurons
Representations of sensory stimuli in the cerebral cortex can undergo progressive remodelling according to the behavioural importance of the stimuli. The cortex receives widespread projections from dopamine neurons in the ventral tegmental area (VTA), which are activated by new stimuli or unpredicted rewards, and are believed to provide a reinforcement signal for such learning-related cortical reorganization. In the primary auditory cortex (AI) dopamine release has been observed during auditory learning that remodels the sound-frequency representations. Furthermore, dopamine modulates long-term potentiation, a putative cellular mechanism underlying plasticity. Here we show that stimulating the VTA together with an auditory stimulus of a particular tone increases the cortical area and selectivity of the neural responses to that sound stimulus in AI. Conversely, the AI representations of nearby sound frequencies are selectively decreased. Strong, sharply tuned responses to the paired tones also emerge in a second cortical area, whereas the same stimuli evoke only poor or non-selective responses in this second cortical field in naive animals. In addition, we found that strong long-range coherence of neuronal discharge emerges between AI and this secondary auditory cortical area.
477754
The locus coeruleus–noradrenergic system: modulation of behavioral state and state-dependent cognitive processes
Through a widespread efferent projection system, the locus coeruleus-noradrenergic system supplies norepinephrine throughout the central nervous system. Initial studies provided critical insight into the basic organization and properties of this system. More recent work identifies a complicated array of behavioral and electrophysiological actions that have in common the facilitation of processing of relevant, or salient, information. This involves two basic levels of action. First, the system contributes to the initiation and maintenance of behavioral and forebrain neuronal activity states appropriate for the collection of sensory information (e.g. waking). Second, within the waking state, this system modulates the collection and processing of salient sensory information through a diversity of concentration-dependent actions within cortical and subcortical sensory, attention, and memory circuits. Norepinephrine-dependent modulation of long-term alterations in synaptic strength, gene transcription and other processes suggest a potentially critical role of this neurotransmitter system in experience-dependent alterations in neural function and behavior. The ability of a given stimulus to increase locus coeruleus discharge activity appears independent of affective valence (appetitive vs. aversive). Combined, these observations suggest that the locus coeruleus-noradrenergic system is a critical component of the neural architecture supporting interaction with, and navigation through, a complex world. These observations further suggest that dysregulation of locus coeruleus-noradrenergic neurotransmission may contribute to cognitive and/or arousal dysfunction associated with a variety of psychiatric disorders, including attention-deficit hyperactivity disorder, sleep and arousal disorders, as well as certain affective disorders, including post-traumatic stress disorder. Independent of an etiological role in these disorders, the locus coeruleus-noradrenergic system represents an appropriate target for pharmacological treatment of specific attention, memory and/or arousal dysfunction associated with a variety of behavioral/cognitive disorders.
24141340
The distribution of tyrosine hydroxylase in the canary brain: demonstration of a specific and sexually dimorphic catecholaminergic innervation of the telencephalic song control nuclei
Abstract. Singing and the processing of auditory information related to song can be affected by experimental manipulations of catecholamine activity in the brain of zebra finches. We investigated, by immunocytochemistry in the brain of male and female canaries, the distribution of tyrosine hydroxylase (TH), the rate-limiting step in the synthesis of catecholamines. Fibers immunoreactive for TH (TH-ir) were particularly abundant in the lobus parolfactorius, the paleostriatum primitivum, and the nucleus septalis lateralis. A high density of TH-ir basket-like structures was observed in the caudomedial neostriatum, an area involved in song perception and recognition. In most males, a high density of TH-ir fibers outlined the telencephalic song control nuclei including the high vocal center, the nucleus robustus archistriatalis, the nucleus interfascialis, the lateral and medial parts of the magnocellular nucleus of the anterior neostriatum, and area X of the lobus parolfactorius. The higher density of fibers immunoreactive for TH in these nuclei, compared with the surrounding telencephalon, supports the notion that the morphological evolution of the song control nuclei was accompanied by a neurochemical specialization. This specific innervation of the song control regions was, in general, not found in females. The specific presence of high densities of TH-ir fibers in the song system of male canaries and the sex difference of this innervation provide anatomical evidence in support of the claim that dopamine and/or norepinephrine play important roles in the modulation of song learning and production.
25541433
Noradrenergic control of auditory information processing in female canaries
An ethological procedure, based on the study of the sexual responsiveness of female canaries (Serinus canaria) to song playbacks was used to investigate the function of central noradrenergic inputs in the processing of auditory information. The effects of a noradrenergic denervation on sexual responses was analyzed in females exposed to playbacks of biological relevant auditory stimuli, i.e. sexually stimulating songs, presented alone or masked by auditory distractors. A decrease in behavioral responsiveness was observed as a function of the amount of masking distractors indicating that female canaries have the perceptual ability to discriminate and selectively attend to biologically relevant songs. After the systemic administration of DSP-4, a specific noradrenergic neurotoxin, females exhibited an overall decrease in sexual responsiveness to songs masked or not by distractors. No effect of DSP-4 were detected on the motor activity nor on reproductive behaviors. These results indicate that central noradrenergic inputs modulate the sexual behavior of female canaries by affecting the auditory processing of relevant information contained in sexually stimulating songs.
24848766
Fos responses of dopamine neurons to sociosexual stimuli in male zebra finches
Dopamine (DA) is produced in numerous brain areas and influences a wide variety of social behaviors, but very few data are available to establish the socially-relevant response properties of most DA populations, which comprise eight cell groups numbered A8-A15. Anatomically, these DA populations are evolutionarily conserved, and all have been identified in both birds and mammals. We now report the Fos responses of tyrosine hydroxylase-immunoreactive (TH-ir; putatively dopaminergic) neurons in the A8-A15 cell groups of male zebra finches following exposure to a control condition or one of six different social stimuli: a heterospecific male, conspecific male, fighting in a mate competition paradigm (which includes both male and female stimuli), a courtship interaction without physical contact, a courtship interaction with physical contact but no mounting, and a courtship interaction with mounting. We found that the DA cell groups exhibit distinctive profiles of responsiveness to social stimuli. Fos induction in A8, A9, A10 and midbrain A11 neurons increased significantly in response to a variety of conspecific stimuli, but not heterospecific stimuli. In contrast, Fos induction in the preoptic A14 neurons was observed specifically in response to sexual interactions, and Fos induction in hypothalamic A11 neurons appears to primarily reflect the performance of courtship singing. Infundibular A12 neurons, which may be involved in stress-related processes, showed the highest level of TH+Fos colocalization in control subjects. This colocalization decreased in response to all conspecific stimuli except fighting, and did not decrease following exposure to a heterospecific male.
36412130
Estradiol modulates brainstem catecholaminergic cell groups and projections to the auditory forebrain in a female songbird
In songbirds, hearing conspecific song induces robust expression of the immediate early gene zenk in the auditory forebrain. This genomic response to song is well characterized in males and females of many species, and is highly selective for behaviorally relevant song. In white-throated sparrows, the selectivity of the zenk response requires breeding levels of estradiol; we previously showed that in non-breeding females with low levels of plasma estradiol, the zenk response to hearing song is no different than the response to hearing frequency-matched tones. Here, we investigated the role of brainstem catecholaminergic cells groups, which project to the forebrain, in estradiol-dependent selectivity. First, we hypothesized that estradiol treatment affects catecholaminergic innervation of the auditory forebrain as well as its possible sources in the brainstem. Immunohistochemical staining of tyrosine hydroxylase revealed that estradiol treatment significantly increased the density of catecholaminergic innervation of the auditory forebrain as well as the number of catecholaminergic cells in the locus coeruleus (A6) and the ventral tegmental area (A10), both of which are known to contain estrogen receptors in songbirds. Second, we hypothesized that during song perception, catecholaminergic cell groups of the brainstem actively participate in auditory selectivity via estrogen-dependent changes in activity. We found that hearing songs did not induce the expression of zenk, a putative marker of activity, within catecholaminergic neurons in any of the cell groups quantified. Together, our results suggest that estradiol induces changes in brainstem catecholaminergic cell groups that may play a neuromodulatory role in behavioral and auditory selectivity.
34053873
Fos-like immunoreactivity in catecholaminergic brain nuclei after territorial behavior in free-living song sparrows.
The catecholaminergic cell groups of the brainstem play an important role in the regulation of motivated behavior, including reproductive behavior. In songbirds, these cell groups project to telencephalic nuclei involved in singing and contain steroid hormone receptors, implicating them in the seasonal regulation of song. Whether these nuclei are involved in the activation of song on a short-term, moment-to-moment basis is unknown. In this study, free-living male song sparrows (Melospiza melodia) were subjected to simulated territorial intrusion (STI), which stimulates territorial singing. The resulting fos-like immunoreactivity (FLI) was quantified in two HVc- and RA-projecting catecholaminergic regions of the brainstem: the area ventralis of Tsai (AVT) and the midbrain central gray (GCt). Males subjected to STI showed more FLI in both of these regions than control males. In addition, FLI in both nuclei was correlated positively with the number of songs sung in response to STI. The number of flights directed at the intruder was correlated with FLI in AVT but not GCt. These results suggest a role for AVT and GCt, and thus possibly catecholamines, in the regulation of territorial behavior in songbirds.
883945
Evidence that norepinephrine influences responses to male courtship song and activity within song control regions and the ventromedial nucleus of the hypothalamus in female European starlings
Little is known about the neural regulation of female responses to male courtship displays. Female European starlings use attributes of male song to assess potential mates and can be highly motivated to approach nest boxes broadcasting male song. The catecholamine norepinephrine (NE) regulates attention and arousal and has recently been implicated in motivated/goal-directed behaviors, suggesting a role for NE in female responses to male song. In the present study, treating females with the noradrenergic neurotoxin N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine hydrochloride (DSP-4) increased the number of times females were observed on or near a nest box broadcasting sexually motivated male song, suggesting an inhibitory role for norepinephrine in the regulation of female responses to male song. DSP-4 treatment reduced immunolabeling for the immediate early gene ZENK within area X, a region involved in song learning and possibly discrimination, and the ventromedial nucleus of the hypothalamus, a region involved in female sexual behavior. Furthermore, female responses to male song and numbers of ZENK-labeled cells within area X related positively in DSP-4-treated birds but not controls. Female responses to male song and the density of dopamine-beta-hydroxylase in area X and another song nucleus LMAN related positively exclusively in DSP-4-treated birds. Similar trends were consistently observed for the ventromedial nucleus of the hypothalamus. These data suggest roles for area X, LMAN, and the ventromedial nucleus of the hypothalamus in female responses to male song and highlight norepinephrine as an important neuromodulator of this behavior.
3243488
What birdsong can teach us about the central noradrenergic system
Increasing evidence indicates that the noradrenergic system plays a key role in biasing the nervous system towards producing behaviors that help animals adapt to constantly changing environments. Most of the studies investigating noradrenergic function are performed in animals that have a limited repertoire of tractable natural behaviors. Songbirds, in contrast, with their rich set of precisely quantifiable vocal behaviors, provide a unique model system to study the noradrenergic system. An additional advantage of this system is the existence of a well-defined neural circuit, known as the song system, that is necessary for the production, learning and perception of song and can be studied at many different levels. These include the ability to investigate the effect of norepinephrine on synaptic function using brain slices, identifying its influence on singing-related gene expression and monitoring its impact on the activity of single neurons recorded in awake behaving birds. In this review article, we describe the similarities and differences, both anatomical and functional, between the avian and mammalian noradrenergic system and its role in sensory processing, learning, attention and synaptic modulation. We also describe how the noradrenergic system influences motor production, an under-explored aspect of norepinephrine function in mammalian studies. We argue that the richness of behaviors observed in songbirds provides a unique opportunity to study the noradrenergic system in a highly integrative manner that will ultimately provide important insights into the role of this system in normal behavior and disease.
24100752
The origin of catecholaminergic inputs to the song control nucleus RA in canaries
&NA; Song control nuclei in oscines receive dense catecholaminergic inputs but their anatomical origin is poorly understood. We analyzed catecholaminergic inputs to the nucleus robustus archistriatalis (RA) in canaries by retrograde tract‐tracing combined with immunocytochemistry for tyrosine hydroxylase. In both sexes, dopaminergic inputs to RA come mostly from the A1 1 (mesencephalic central gray) and A10 (area ventralis of Tsai) cell groups but the locus coeruleus and subcoeruleus (A6) also send noradrenergic projections to RA. No input originates in the hypothalamic and in the A5 to A1 catecholaminergic groups. These findings and previous work on the high vocal center (HVc) indicate that the two major nuclei of the motor pathway controlling song production (RA and HVc) receive catecholaminergic inputs of similar origins.
37062944
Noradrenergic Deficits Alter Processing of Communication Signals in Female Songbirds
During acoustic communication, animals must attend to sounds from a particular source while simultaneously rejecting intrusion from other sources. One possible candidate mechanism for this process is the noradrenergic system. Noradrenaline is a neuromodulator that tunes sensory processing systems and regulates attention. We examined whether pharmacological degradation of the noradrenergic system using N-(2-chloroethyl)-N-2-bromobenzyl-amine hydrochloride (DSP-4) modifies processing of species-typical auditory signals in female canaries (Serinus canaria). We measured auditory responses to conspecific and heterospecific songs using ZENK protein expression within the caudomedial nidopallium (NCM) and the mesopallium caudomedial (CMM). Song-induced ZENK expression in these auditory forebrain areas is typically higher in birds exposed to conspecific songs as opposed to heterospecific songs. Our results reveal that this differential ZENK induction is abolished specifically within dNCM and CMM in female canaries treated with DSP-4. Furthermore, in DSP-4-treated birds, conspecific song-induced ZENK expression is significantly reduced when compared to saline-treated birds. This suggests that the noradrenergic system modifies auditory processing by enhancing neuronal responses to signals relevant to survival and reproduction rather than inhibiting neuronal responses to signals that are less relevant. Overall, our results reveal that noradrenaline plays a significant neuromodulatory role during the reception of species-typical communication signals.
26097583
Noradrenergic neurotoxin, N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine hydrochloride (DSP-4), treatment eliminates estrogenic effects on song responsiveness in female zebra finches (Taeniopygia guttata).
Female songbirds use male songs as an important criterion for mate selection. Several studies have reported that female songbirds prefer complex songs to other song types. In a recent study, the authors found that song responsiveness in female zebra finches (Taeniopygia guttata) is strongly modulated by circulating estrogen levels. The behavioral effects of estrogen are often mediated via norepinephrine (NE). The current study administered the noradrenergic neurotoxin, N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine hydrochloride (DSP-4) to estradiol-treated female zebra finches to investigate if estrogenic effects on song responsiveness are mediated via NE. The authors tested song responsiveness of adult female zebra finches for three acoustically different song types--simple, long-bout, and complex--under three treatment conditions, untreated, estradiol-treated, and estradiol + DSP-4-treated. Females only showed differential song responsiveness when treated with estradiol alone, responding more to complex songs. DSP-4 treatment eliminated this differential responsiveness. The results are discussed in the light of evidence from functional, neurochemical, and neuroanatomical studies that suggest that estrogenic effects on song processing might be mediated by NE.
33965450
Selective behavioral responses to male song are affected by the dopamine agonist GBR-12909 in female European starlings (Sturnus vulgaris)
Female songbirds use attributes of male song to select mates. Different types of male song differ in incentive value (or the ability to attract females). Dopamine plays a role in incentive value and reward; however, little is known about its role in selective female behavioral responses to male courtship signals. We examined the effects of the indirect dopamine agonist (dopamine reuptake inhibitor) GBR-12909 on female songbird responses to male song stimuli. Female European starlings were played recordings of long starling song (presumed high incentive value), short starling song (presumed lower incentive value), or purple martin song (lowest incentive value). Vehicle-treated females investigated nest boxes playing starling song more than purple martin song. However, GBR-12909 disrupted preferential responses to the starling song stimuli. GBR-12909 also increased cFOS immunolabeling in the ventromedial nucleus of the hypothalamus (VMH) at the same dose that disrupted female selective responses to male starling song. The results suggest that dopamine receptors play an important role in female selective responses to biologically meaningful stimuli and that the VMH may be influenced by dopamine to alter female responses to male song.
7335731
α1-Noradrenegic receptor antagonism disrupts female songbird responses to male song
Female songbirds respond behaviorally to differences in male song structure. Past data suggest a complex role for norepinephrine in female responses to song. Here, we examined the effects of central infusions of the α(1)-noradrenergic receptor antagonist prazosin on female European starling (Sturnus vulgaris) responses to nest boxes broadcasting male song. Prazosin disrupted female preferential responses to male starling song over the less biologically relevant purple martin (Progne subis) song. Prazosin decreased female responses to male starling song in a linear dose-response fashion; whereas, it affected responses to purple martin song in a U-shaped dose-response fashion. Results suggest that the role of norepinephrine in female responses to male song differs depending upon drug dose and the biological relevance of the song stimulus.
17812483
Estradiol‐dependent catecholaminergic innervation of auditory areas in a seasonally breeding songbird
A growing body of evidence suggests that gonadal steroids such as estradiol (E2) alter neural responses not only in brain regions associated with reproductive behavior but also in sensory areas. Because catecholamine systems are involved in sensory processing and selective attention, and because they are sensitive to E2 in many species, they may mediate the neural effects of E2 in sensory areas. Here, we tested the effects of E2 on catecholaminergic innervation, synthesis and activity in the auditory system of white‐throated sparrows, a seasonally breeding songbird in which E2 promotes selective auditory responses to song. Non‐breeding females with regressed ovaries were held on a winter‐like photoperiod and implanted with silastic capsules containing either no hormone or E2. In one hemisphere of the brain, we used immunohistochemistry to quantify fibers immunoreactive for tyrosine hydroxylase or dopamine beta‐hydroxylase in the auditory forebrain, thalamus and midbrain. E2 treatment increased catecholaminergic innervation in the same areas of the auditory system in which E2 promotes selectivity for song. In the contralateral hemisphere we quantified dopamine, norepinephrine and their metabolites in tissue punches using HPLC. Norepinephrine increased in the auditory forebrain, but not the midbrain, after E2 treatment. We found that evidence of interhemispheric differences, both in immunoreactivity and catecholamine content that did not depend on E2 treatment. Overall, our results show that increases in plasma E2 typical of the breeding season enhanced catecholaminergic innervation and synthesis in some parts of the auditory system, raising the possibility that catecholamines play a role in E2‐dependent auditory plasticity in songbirds.
17342068
The integration of song environment by catecholaminergic systems innervating the auditory telencephalon of adult female European starlings
Mate choice is among the most consequential decisions a sexually reproducing organism can make. In many songbird species, females make mate‐choice decisions based, in part, on variation between males in songs that reflect their quality. Importantly, females may adjust their choice relative to the prevalence of high quality songs. In European starlings (Sturnus vulgaris), females prefer males that primarily sing long songs over those that primarily sing short songs, and sensitivity of the auditory telencephalon to song length depends on the prevalence of long songs in the environment. Several lines of evidence suggest a role for noradrenergic innervation of the auditory telencephalon in mediating this neuro‐ and behavioral plasticity. To simulate variation in quality of the song environment, we exposed adult female starlings to 1 week of either long or short songs and then quantified several monoamines and their metabolites in the caudomedial mesopallium and caudomedial nidopallium (NCM) using high performance liquid chromatography. We also used immunocytochemistry to assess these areas for immunoreactive dopamine‐β‐hydroxylase (DBH‐ir), the enzyme that synthesizes norepinephrine. We found that long songs elevated levels of the principal norepinephrine metabolite, the principal dopamine metabolite, and the probability of DBH‐ir in the NCM compared to short songs. Song environment did not appear to influence norepinephrine or dopamine levels. Thus, the quality of the song environment regulates the local secretion of catecholamines, particularly norepinephrine, in the female auditory telencephalon. This may form a basis for plasticity in forebrain sensitivity and mate‐choice behavior based on the prevalence of high‐quality males. © 2008 Wiley Periodicals, Inc. Develop Neurobiol 2008
46371345
Phasic versus tonic dopamine release and the modulation of dopamine system responsivity: A hypothesis for the etiology of schizophrenia
A novel mechanism for regulating dopamine activity in subcortical sites and its possible relevance to schizophrenia is proposed. This hypothesis is based on the regulation of dopamine release into subcortical regions occurring via two independent mechanisms: (1) transient or phasic dopamine release caused by dopamine neuron firing, and (2) sustained, "background" tonic dopamine release regulated by prefrontal cortical afferents. Behaviorally relevant stimuli are proposed to cause short-term activation of dopamine cell firing to trigger the phasic component of dopamine release. In contrast, tonic dopamine release is proposed to regulate the intensity of the phasic dopamine response through its effect on extracellular dopamine levels. In this way, tonic dopamine release would set the background level of dopamine receptor stimulation (both autoreceptor and postsynaptic) and, through homeostatic mechanisms, the responsivity of the system to dopamine in these sites. In schizophrenics, a prolonged decrease in prefrontal cortical activity is proposed to reduce tonic dopamine release. Over time, this would elicit homeostatic compensations that would increase overall dopamine responsivity and thereby cause subsequent phasic dopamine release to elicit abnormally large responses.
31352084
The monoamine innervation of rat cerebral cortex: Synaptic and nonsynaptic axon terminals
Publisher Summary This chapter discusses the monoamine innervations of rat cerebral cortex. Substantial evidence recently obtained from experiments carried out on the monoaminergic systems of the mammalian brain indicates that molecular communication between nerve cells may take various forms differing from the traditional views on synaptic transmission. Another new concept evolved from the notion that in certain parts of the central nervous system, a high proportion of monoamine axon terminals lacks the membranous apposition and specializations that characterize synapses in electron micrographs. This finding led to the suggestion that the biogenic amines released from such nonsynaptic nerve endings might reach remote receptors: A mode of operation intermediate between the private addressing of classical synaptic messengers and the broadcasting of neuroendocrine secretion. The chapter presents the main body of radioautographic data that gave rise to this hypothesis and discusses some of their cytological and functional implications.
22641937
Modulatory role for biogenic amines in the cerebral cortex. Microiontophoretic studies
In order to investigate the mode of action of biogenic amines in rat cerebral cortex, the unitary activity of spontaneously firing neurons and their excitatory response to acetylcholine (ACh) were examined using microiontophoretic administration of dopamine (DA), noradrenaline (NA) and serotonin (5-HT). The predominant effect of these biogenic amines on the spontaneous activity was a profound and prolonged inhibition of firing (2-4 min), which attained its maximum within 15-120 sec. This response was generally more abrupt in onset and of greater magnitude with NA and 5-HT than with DA. Most units inhibited by DA, NA and 5-HT also showed marked depression of their excitatory response to ACh when pretreated with these biogenic amines. With repetitive administration of ACh, it could be shown that the total duration of inhibition of ACh responses by DA and NA was not as prolonged as the inhibition of the spontaneous firing of the same cells. With 5-HT, the initial ACh responses of many neurons could be completely blocked, and this inhibitory effect lasted as long as the inhibition of spontaneous firing. In view of the anatomical data demonstrating a relative sparsity of monoamine nerve terminals in cerebral cortex, the strong inhibition induced by DA, NA or 5-HT may have reflected slow inactivation of the biogenic amines. However, it could also be indicative of underlying mechanisms of action dependent on metabolic changes. Indeed, the interaction between biogenic amines and ACh might imply a balance between the intracellular pools of cAMP and cGMP is directly or indirectly influenced by the biogenic amines and ACh, respectively. This hypothesis would not exclude other modes of local interaction between DA, NA, 5-HT and ACh, and appears compatible with the modulatory role of biogenic amines in cerebral cortex.
13503219
Multiple dopamine functions at different time courses.
Many lesion studies report an amazing variety of deficits in behavioral functions that cannot possibly be encoded in great detail by the relatively small number of midbrain dopamine neurons. Although hoping to unravel a single dopamine function underlying these phenomena, electrophysiological and neurochemical studies still give a confusing, mutually exclusive, and partly contradictory account of dopamine's role in behavior. However, the speed of observed phasic dopamine changes varies several thousand fold, which offers a means to differentiate the behavioral relationships according to their time courses. Thus dopamine is involved in mediating the reactivity of the organism to the environment at different time scales, from fast impulse responses related to reward via slower changes with uncertainty, punishment, and possibly movement to the tonic enabling of postsynaptic motor, cognitive, and motivational systems deficient in Parkinson's disease.
29874665
Convergent regulation of locus coeruleus activity as an adaptive response to stress.
Although hypothalamic-pituitary-adrenal axis activation is generally considered to be the hallmark of the stress response, many of the same stimuli that initiate this response also activate the locus coeruleus-norepinephrine system. Given its functional attributes, the parallel engagement of the locus coeruleus-norepinephrine system with the hypothalamic-pituitary-adrenal axis serves to coordinate endocrine and cognitive limbs of the stress response. The elucidation of stress-related afferents to the locus coeruleus and the electrophysiological characterization of these inputs are revealing how the activity of this system is fine-tuned by stressors to facilitate adaptive cognitive responses. Emerging from these studies, is a picture of complex interactions between the stress-related neuropeptide, corticotropin-releasing factor (CRF), endogenous opioids and the excitatory amino acid neurotransmitter, glutamate. The net effect of these interactions is to adjust the activity and reactivity of the locus coeruleus-norepinephrine system such that state of arousal and processing of sensory stimuli are modified to facilitate adaptive behavioral responses to stressors. This review begins with an introduction to the basic anatomical and physiological characteristics of locus coeruleus neurons. The concept that locus coeruleus neurons operate through two activity modes, i.e., tonic vs. phasic, that determine distinct behavioral strategies is emphasized in light of its relevance to stress. Anatomical and physiological evidence are then presented suggesting that interactions between stress-related neurotransmitters that converge on locus coeruleus neurons regulate shifts between these modes of discharge in response to the challenge of a stressor. This review focuses specifically on the locus coeruleus because it is the major source of norepinephrine to the forebrain and has been implicated in behavioral and cognitive aspects of stress responses.
11794383
The Genomic Action Potential
Neurons compute in part by integrating, on a time scale of milliseconds, many synaptic inputs and generating a digital output-the "action potential" of classic electrophysiology. Recent discoveries indicate that neurons also perform a second, much slower, integration operating on a time scale of minutes or even hours. The output of this slower integration involves a pulse of gene expression which may be likened to the electrophysiological action potential. Its function, however, is not directed toward immediate transmission of a synaptic signal but rather toward the experience-dependent modification of the underlying synaptic circuitry. Commonly termed the "immediate early gene" (IEG) response, this phenomenon is often assumed to be a necessary component of a linear, deterministic cascade of memory consolidation. Critical review of the large literature describing the phenomenon, however, leads to an alternative model of IEG function in the brain. In this alternative, IEG activation is not directed at the consolidation of memories of a specific inducing event; instead, it sets the overall gain or efficiency of memory formation and directs it to circuits engaged by behaviorally significant contexts. The net result is a sharpening of the selectivity of memory formation, a recruitment of temporally correlated associations, and an ultimate enhancement of long-term memory retrieval.
17391418
Increased Fos expression among midbrain dopaminergic cell groups during birdsong tutoring
During avian vocal learning, birds memorize conspecific song patterns and then use auditory feedback to match their vocal output to this acquired template. Some models of song learning posit that during tutoring, conspecific visual, social and/or auditory cues activate neuromodulatory systems that encourage acquisition of the tutor’s song and attach incentive value to that specific acoustic pattern. This hypothesis predicts that stimuli experienced during social tutoring activate cell populations capable of signaling reward. Using immunocytochemistry for the protein product of the immediate early gene c‐Fos, we found that brief exposure of juvenile male zebra finches to a live familiar male tutor increased the density of Fos+ cells within two brain regions implicated in reward processing: the ventral tegmental area (VTA) and substantia nigra pars compacta (SNc). This activation of Fos appears to involve both dopaminergic and non‐dopaminergic VTA/SNc neurons. Intriguingly, a familiar tutor was more effective than a novel tutor in stimulating Fos expression within these regions. In the periaqueductal gray, a dopamine‐enriched cell population that has been implicated in emotional processing, Fos labeling also was increased after tutoring, with a familiar tutor again being more effective than a novel conspecific. As several neural regions implicated in song acquisition receive strong dopaminergic projections from these midbrain nuclei, their activation in conjunction with hearing the tutor’s song could help to establish sensory representations that later guide motor sequence learning.
207515295
A Basal Ganglia Pathway Drives Selective Auditory Responses in Songbird Dopaminergic Neurons via Disinhibition
Dopaminergic neurons in mammals respond to rewards and reward-predicting cues, and are thought to play an important role in learning actions or sensory cues that lead to reward. The anatomical sources of input that drive or modulate such responses are not well understood; these ultimately define the range of behavior to which dopaminergic neurons contribute. Primary rewards are not the immediate objective of all goal-directed behavior. For example, a goal of vocal learning is to imitate vocal-communication signals. Here, we demonstrate activation of dopaminergic neurons in songbirds driven by a basal ganglia region required for vocal learning, area X. Dopaminergic neurons in anesthetized zebra finches respond more strongly to the bird's own song (BOS) than to other sounds, and area X is critical for these responses. Direct pharmacological modulation of area X output, in the absence of auditory stimulation, is sufficient to bidirectionally modulate the firing rate of dopaminergic neurons. The only known pathway from song control regions to dopaminergic neurons involves a projection from area X to the ventral pallidum (VP), which in turn projects to dopaminergic regions. We show that VP neurons are spontaneously active and inhibited preferentially by BOS, suggesting that area X disinhibits dopaminergic neurons by inhibiting VP. Supporting this model, auditory-response latencies are shorter in area X than VP, and shorter in VP than dopaminergic neurons. Thus, dopaminergic neurons can be disinhibited selectively by complex sensory stimuli via input from the basal ganglia. The functional pathway we identify may allow dopaminergic neurons to contribute to vocal learning.
15228354
Song- and Order-Selective Neurons in the Songbird Anterior Forebrain and their Emergence during Vocal Development
Auditory experience is critical for vocal learning in songbirds as in humans. Therefore, in a search for neural mechanisms for song learning and recognition, the auditory response properties of neurons in the anterior forebrain (AF) pathway of the songbird brain were investigated. This pathway plays an essential but poorly understood role during the period of song development when auditory feedback is most crucial. Single-unit recordings demonstrated that both the lateral magnocellular nucleus of the anterior neostriatum (LMAN) and Area X (X) contain auditory neurons in adult male finches. These neurons are strongly selective for both spectral and temporal properties of song; they respond more robustly to the bird’s own song (BOS) than to songs of conspecific individuals, and they respond less well to the BOS if it is played in reverse. In addition, X neurons are more broadly responsive than LMAN neurons, suggesting that responses to song become progressively more refined along this pathway. Both X and LMAN of young male finches early in the process of song learning (30–45 d old) also contain song-responsive auditory neurons, but these juvenile neurons lack the song and order selectivity present in adult birds. The spectral and temporal selectivity of the adult AF auditory neurons therefore arises during development in neurons that are initially broadly song-responsive. These neurons provide one of the clearest examples of experience-dependent acquisition of complex stimulus selectivity. Moreover, the auditory properties of the AF circuit suggest that one of its functions may be to mediate the auditory learning and feedback so essential to song development.
15400577
Functional organization of forebrain pathways for song production and perception.
This article reviews the organization of the forebrain nuclei of the avian song system. Particular emphasis is placed on recent physiologic recordings from awake behaving adult birds while they sing, call, and listen to broadcasts of acoustic stimuli. The neurons in the descending motor pathway (HVc and RA) are organized in a hierarchical arrangement of temporal units of song production, with HVc neurons representing syllables and RA neurons representing notes. The nuclei Uva and NIf, which are afferent to HVc, may help organize syllables into larger units of vocalization. HVc and RA are also active during production of all calls. The patterns of activity associated with calls differ between learned calls and those that are innately specified, and give insight into the interactions between the forebrain and midbrain during calling, as well as into the evolutionary origins of the song system. Neurons in Area X, the first part of the anterior forebrain pathway leading from HVc to RA, are also active during singing. Many HVc neurons are also auditory, exhibiting selectivity for learned acoustic parameters of the individual bird's own song (BOS). Similar auditory responses are also observed in RA and Area X in anesthetized birds. In contrast to HVc, however, auditory responses in RA are very weak or absent in awake birds under our experimental paradigm, but are uncovered when birds are anesthetized. Thus, the roles of both pathways beyond HVc in adult birds is under review. In particular, theories hypothesizing a role for the descending motor pathway (RA and below) in adult song perception do not appear to obtain. The data also suggest that the anterior forebrain pathway has a greater motor role than previously considered. We suggest that a major role of the anterior forebrain pathway is to resolve the timing mismatch between motor program readout and sensory feedback, thereby facilitating motor programming during birdsong learning. Pathways afferent to HVc may participate more in sensory acquisition and sensorimotor learning during song development than is commonly assumed.
11031112
Immunohistochemical studies on phosphorylation of tyrosine hydroxylase in central catecholamine neurons using site- and phosphorylation state-specific antibodies.
Antibodies raised to phosphorylated forms of tyrosine hydroxylase, the first and rate-limiting enzyme in the catecholamine biosynthesis, were applied in immunohistochemical studies on rat brain slices incubated in vitro with a phosphodiesterase inhibitor (3-isobutyl-1-methylxanthine, IBMX) and on forskolin on formalin-perfused rat brains. Four antisera/antibodies were used: polyclonal rabbit antisera to (i) tyrosine hydroxylase phosphorylated at serine 40 (THS40p antiserum), (ii) tyrosine hydroxylase phosphorylated at serine 19 (THS19p antiserum), (iii) the native enzyme (pan-tyrosine hydroxylase antiserum), and mouse monoclonal antibody to (iv) native tyrosine hydroxylase. In the in vitro studies THS40p-like immunoreactivity was not observed unless slices were treated with IBMX-forskolin after which a dense fibre network was found in the striatum, and immunoreactive cell bodies were found in the ventral mesencephalon, especially in the ventral tegmental area. Although these cells were pan-tyrosine hydroxylase-positive, several of them were not stained with the tyrosine hydroxylase-monoclonal antibody. Moreover, there was a marked reduction of tyrosine hydroxylase-monoclonal antibody-immunoreactive fibres in drug-treated slices, suggesting that this tyrosine hydroxylase-monoclonal antibody does not recognize the serine 40-phosphorylated form of tyrosine hydroxylase. Treated slices did not show any THS40p-immunoreactive cell bodies in the dopaminergic A11 cell group and only a few, weakly fluorescent neurons were observed in locus coeruleus. However, a sparse fibre plexus was observed in locus coeruleus, possibly reflecting epinephrine fibres. In the perfused brains THS40p-like immunoreactivity could be visualized in some dopamine neurons in the ventral mesencephalon, especially the A10 area, and in noradrenergic locus coeruleus neurons, whereas THS19p-like immunoreactivity was found in all catecholamine groups studied, similar to the results obtained with the pan-tyrosine hydroxylase antiserum and the tyrosine hydroxylase-monoclonal antibody. In forebrain areas known to be innervated by mesencephalic dopamine neurons, no THS40p-positive fibres were observed, whereas THS19p-immunoreactive fibres were found in subregions of the striatum, olfactory tubercle and nucleus accumbens, essentially overlapping with dopamine fibres previously shown to contain cholecystokinin-like immunoreactivity. The present results suggests that antibodies directed against phosphorylated forms of tyrosine hydroxylase can be used to evaluate the state of tyrosine hydroxylase phosphorylation in individual neuronal cell bodies and processes both in vitro and in vivo.
38567478
Regulatory mechanism of tyrosine hydroxylase activity.
Activity of tyrosine hydroxylase is regulated by feedback inhibition and inactivation by catecholamines, and activation by protein phosphorylation. In this article, reaction mechanisms for the conversion of tyrosine hydroxylase to an inactive/stable form by catecholamines, and activation of tyrosine hydroxylase by phosphorylation at Ser-40 are discussed. Inactivation may be induced by sub-stoichiometric amounts of catecholamines, and activation by phosphorylation of Ser-40 may require phosphorylation of three or all four subunits of a tyrosine hydroxylase molecule. Cooperative phosphorylation at Ser-40 in the subunits is also discussed.
27886873
Intricate Regulation of Tyrosine Hydroxylase Activity and Gene Expression
Abstract: Tyrosine hydroxylase catalyzes the rate‐limiting step in the biosynthesis of the catecholamines dopamine, norepinephrine, and epinephrine. Therefore, the regulation of tyrosine hydroxylase enzyme number and intrinsic enzyme activity represents the central means for controlling the synthesis of these important biogenic amines. An intricate scheme has evolved whereby tyrosine hydroxylase activity is modulated by nearly every documented form of regulation. Beginning with the genomic DNA, evidence exists for the transcriptional regulation of tyrosine hydroxylase mRNA levels, alternative RNA processing, and the regulation of RNA stability. There is also experimental support for the role of both translational control and enzyme stability in establishing steady‐state levels of active tyrosine hydroxylase protein. Finally, mechanisms have been proposed for feedback inhibition of the enzyme by catecholamine products, allosteric modulation of enzyme activity, and phosphorylation‐dependent activation of the enzyme by various different kinase systems. Given the growing literature suggesting that different tissues regulate tyrosine hydroxylase mRNA levels and activity in different ways, regulatory mechanisms provide not only redundancy but also diversity in the control of catecholamine biosynthesis.
24324381
Tyrosine hydroxylase phosphorylation: regulation and consequences
The rate‐limiting enzyme in catecholamine synthesis is tyrosine hydroxylase. It is phosphorylated at serine (Ser) residues Ser8, Ser19, Ser31 and Ser40 in vitro, in situ and in vivo. A range of protein kinases and protein phosphatases are able to phosphorylate or dephosphorylate these sites in vitro. Some of these enzymes are able to regulate tyrosine hydroxylase phosphorylation in situ and in vivo but the identity of the kinases and phosphatases is incomplete, especially for physiologically relevant stimuli. The stoichiometry of tyrosine hydroxylase phosphorylation in situ and in vivo is low. The phosphorylation of tyrosine hydroxylase at Ser40 increases the enzyme's activity in vitro, in situ and in vivo. Phosphorylation at Ser31 also increases the activity but to a much lesser extent than for Ser40 phosphorylation. The phosphorylation of tyrosine hydroxylase at Ser19 or Ser8 has no direct effect on tyrosine hydroxylase activity. Hierarchical phosphorylation of tyrosine hydroxylase occurs both in vitro and in situ, whereby the phosphorylation at Ser19 increases the rate of Ser40 phosphorylation leading to an increase in enzyme activity. Hierarchical phosphorylation depends on the state of the substrate providing a novel form of control of tyrosine hydroxylase activation.
16225
Altered Dopamine Signaling in Naturally Occurring Maternal Neglect
Background Child neglect is the most common form of child maltreatment, yet the biological basis of maternal neglect is poorly understood and a rodent model is lacking. Methodology/Principal Findings The current study characterizes a population of mice (MaD1) which naturally exhibit maternal neglect (little or no care of offspring) at an average rate of 17% per generation. We identified a set of risk factors that can predict future neglect of offspring, including decreased self-grooming and elevated activity. At the time of neglect, neglectful mothers swam significantly more in a forced swim test relative to nurturing mothers. Cross-fostered offspring raised by neglectful mothers in turn exhibit increased expression of risk factors for maternal neglect and decreased maternal care as adults, suggestive of possible epigenetic contributions to neglect. Unexpectedly, offspring from neglectful mothers elicited maternal neglect from cross-fostered nurturing mothers, suggesting that factors regulating neglect are not solely within the mother. To identify a neurological pathway underlying maternal neglect, we examined brain activity in neglectful and nurturing mice. c-Fos expression was significantly elevated in neglectful relative to nurturing mothers in the CNS, particularly within dopamine associated areas, such as the zona incerta (ZI), ventral tegmental area (VTA), and nucleus accumbens. Phosphorylated tyrosine hydroxylase (a marker for dopamine production) was significantly elevated in ZI and higher in VTA (although not significantly) in neglectful mice. Tyrosine hydroxylase levels were unaltered, suggesting a dysregulation of dopamine activity rather than cell number. Phosphorylation of DARPP-32, a marker for dopamine D1-like receptor activation, was elevated within nucleus accumbens and caudate-putamen in neglectful versus nurturing dams. Conclusions/Significance These findings suggest that atypical dopamine activity within the maternal brain, especially within regions involved in reward, is involved in naturally occurring neglect and that MaD1 mice are a useful model for understanding the basis of naturally occurring neglect.
34990309
Evidence that female endocrine state influences catecholamine responses to male courtship song in European starlings.
Little is known about the neural control of female responses to male courtship. Female European starlings in breeding condition with high concentrations of estrogen select mates based on variation in song and approach nest boxes broadcasting male song. In contrast, outside of the breeding season (when estrogen is low) females do not display the same response to male song. The catecholamines dopamine and norepinephrine regulate behaviors important for mate choice such as arousal, attention, sexual motivation, and goal-directed approach responses, suggesting a role for catecholamines in female responses to male song. In the present study, treating females with a dopamine agonist inhibited, whereas an antagonist stimulated female interest in nest boxes broadcasting male song. In a second study immunocytochemistry was used to examine the distribution of the phosphorylated (i.e., active) form of tyrosine hydroxylase (pTH), the rate-limiting enzyme for catecholamine synthesis. Exposure to male song in breeding condition females reduced pTH density in brain regions involved in social behavior (lateral septum, ventromedial nucleus of the hypothalamus) and a region involved in visual processing (nucleus of Edinger-Westphal) but not song control regions. Opposite patterns of pTH labeling densities were observed in the same regions in response to song in non-breeding condition females. pTH in the ventral tegmental area was also affected by song and female endocrine condition. Overall, the present data support an inhibitory role for dopamine in female responses to courtship and suggest that endocrine state and catecholamines interact to regulate this behavior.
45928478
Estradiol-dependent modulation of auditory processing and selectivity in songbirds
The steroid hormone estradiol plays an important role in reproductive development and behavior and modulates a wide array of physiological and cognitive processes. Recently, reports from several research groups have converged to show that estradiol also powerfully modulates sensory processing, specifically, the physiology of central auditory circuits in songbirds. These investigators have discovered that (1) behaviorally-relevant auditory experience rapidly increases estradiol levels in the auditory forebrain; (2) estradiol instantaneously enhances the responsiveness and coding efficiency of auditory neurons; (3) these changes are mediated by a non-genomic effect of brain-generated estradiol on the strength of inhibitory neurotransmission; and (4) estradiol regulates biochemical cascades that induce the expression of genes involved in synaptic plasticity. Together, these findings have established estradiol as a central regulator of auditory function and intensified the need to consider brain-based mechanisms, in addition to peripheral organ dysfunction, in hearing pathologies associated with estrogen deficiency.
6173245
The organization of the ascending auditory pathway in the pigeon (Columba livia). I. Diencephalic projections of the inferior colliculus (nucleus mesencephali lateralis, pars dorsalis).
Abstract 1. An experimental-anatomical study of the projection from the inferior colliculus (MLD) in the pigeon was performed by the aid of the Nauta-Gygax method. The resulting observations can be summarized as follows. 2. A massive efferent fiber system — the brachium of the inferior colliculus—ascends from the MLD, passing medial to the brachium of the superior colliculus and dorsal to the ectomammillary nucleus. 3. The majority of its fibers are distributed to the ipsilateral nucleus ovoidalis via the prominent tractus nuclei ovoidalis. 4. Other fibers from the brachium of the inferior colliculus enter the dorsal supraoptic decussation (DSOD), terminating in passage on neurons of the nucleus of the DSOD, and join the contralateral tractus nuclei ovoidalis to end in the nucleus ovoidalis of the opposite side. 5. Degenerating axons were followed to the contralateral MLD and, via a descending path, to the ipsilateral trapezoid body. 6. No axonal degeneration was seen in nucleus ovoidalis after lesions of the spinal cord, nuclei cuneatus and gracilis, nucleus solitarius, trigeminal nucleus, cerebellar nuclei, retina or optic tectum. 7. It is concluded that nucleus ovoidalis represents a specific diencephalic auditory relay nucleus, possibly comparable to the ventral division (Morest) of the mammalian medial geniculate nucleus.
12126302
Role of the zebra finch auditory thalamus in generating complex representations for natural sounds.
We estimated the spectrotemporal receptive fields of neurons in the songbird auditory thalamus, nucleus ovoidalis, and compared the neural representation of complex sounds in the auditory thalamus to those found in the upstream auditory midbrain nucleus, mesencephalicus lateralis dorsalis (MLd), and the downstream auditory pallial region, field L. Our data refute the idea that the primary sensory thalamus acts as a simple, relay nucleus: we find that the auditory thalamic receptive fields obtained in response to song are more complex than the ones found in the midbrain. Moreover, we find that linear tuning diversity and complexity in ovoidalis (Ov) are closer to those found in field L than in MLd. We also find prevalent tuning to intermediate spectral and temporal modulations, a feature that is unique to Ov. Thus even a feed-forward model of the sensory processing chain, where neural responses in the sensory thalamus reveals intermediate response properties between those in the sensory periphery and those in the primary sensory cortex, is inadequate in describing the tuning found in Ov. Based on these results, we believe that the auditory thalamic circuitry plays an important role in generating novel complex representations for specific features found in natural sounds.
205430768
Tuning for spectro-temporal modulations as a mechanism for auditory discrimination of natural sounds
Vocal communicators discriminate conspecific vocalizations from other sounds and recognize the vocalizations of individuals. To identify neural mechanisms for the discrimination of such natural sounds, we compared the linear spectro-temporal tuning properties of auditory midbrain and forebrain neurons in zebra finches with the statistics of natural sounds, including song. Here, we demonstrate that ensembles of auditory neurons are tuned to auditory features that enhance the acoustic differences between classes of natural sounds, and among the songs of individual birds. Tuning specifically avoids the spectro-temporal modulations that are redundant across natural sounds and therefore provide little information; rather, it overlaps with the temporal modulations that differ most across sounds. By comparing the real tuning and a less selective model of spectro-temporal tuning, we found that the real modulation tuning increases the neural discrimination of different sounds. Additionally, auditory neurons discriminate among zebra finch song segments better than among synthetic sound segments.
39830927
Auditory pathways of caudal telencephalon and their relation to the song system of adult male zebra finches (Taenopygia guttata)
Auditory information is critical for vocal imitation and other elements of social life in songbirds. In zebra finches, neural centers that are necessary for the acquisition and production of learned vocalizations are known, and they all respond to acoustic stimulation. However, the circuits by which conspecific auditory signals are perceived, processed, and stored in long‐term memory have not been well documented. In particular, no evidence exists of direct connections between auditory and vocal motor pathways, and two newly identified centers for auditory processing, caudomedial neostriatum (Ncm) and caudomedial hyperstriatum ventrale (cmHV), have no documented place among known auditory circuits. Our goal was to describe anatomically the auditory pathways in adult zebra finch males and, specifically, to show the projections by which Ncm and vocal motor centers may receive auditory input. By using injections of different kinds of neuroanatomical tracers (biotinylated dextran amines, rhodamine‐linked dextran amines, biocytin, fluorogold, and rhodamine‐linked latex beads), we have shown that, as in other avian groups, the neostriatal field L complex in caudal telencephalon is the primary forebrain relay for pathways originating in the auditory thalamus, i.e., the nucleus ovoidalis complex (Ov). In addition, Ncm and cmHV also receive input from the Ov complex. Ov has been broken down into two parts, the Ov “core” and “shell,” which project in parallel to different targets in the caudal telencephalon. Parts of the field L complex are connected among themselves and to Ncm, cmHV, and caudolateral HV (cIHV) through a complex web of largely reciprocal pathways. In addition, cIHV and parts of the field L complex project strongly to the “shelf” of neostriatum underneath the song control nucleus high vocal center (HVC) and to the “cup” of archistriatum rostrodorsal to another song‐control nucleus, the robust nucleus of the archistriatum (RA). We have documented two points at which the vocal motor pathway may pick up auditory signals: the HVC‐shelf interface and a projection from cIHV to the nucleus interfacialis (NIf), which projects to HVC. These data represent the most complete survey to date of auditory pathways in the adult male zebra finch brain, and of their projections to motor stations of the song system. © 1996 Wiley‐Liss, Inc.
16607208
A songbird forebrain area potentially involved in auditory discrimination and memory formation
Songbirds rely on auditory processing of natural communication signals for a number of social behaviors, including mate selection, individual recognition and the rare behavior of vocal learning — the ability to learn vocalizations through imitation of an adult model, rather than by instinct. Like mammals, songbirds possess a set of interconnected ascending and descending auditory brain pathways that process acoustic information and that are presumably involved in the perceptual processing of vocal communication signals. Most auditory areas studied to date are located in the caudomedial forebrain of the songbird and include the thalamo-recipient field L (subfields L1, L2 and L3), the caudomedial and caudolateral mesopallium (CMM and CLM, respectively) and the caudomedial nidopallium (NCM). This review focuses on NCM, an auditory area previously proposed to be analogous to parts of the primary auditory cortex in mammals. Stimulation of songbirds with auditory stimuli drives vigorous electrophysiological responses and the expression of several activity-regulated genes in NCM. Interestingly, NCM neurons are tuned to species-specific songs and undergo some forms of experience-dependent plasticity in-vivo. These activity-dependent changes may underlie long-term modifications in the functional performance of NCM and constitute a potential neural substrate for auditory discrimination. We end this review by discussing evidence that suggests that NCM may be a site of auditory memory formation and/or storage.
30528951
Estradiol Shapes Auditory Processing in the Adult Brain by Regulating Inhibitory Transmission and Plasticity-Associated Gene Expression
Estradiol impacts a wide variety of brain processes, including sex differentiation, mood, and learning. Here we show that estradiol regulates auditory processing of acoustic signals in the vertebrate brain, more specifically in the caudomedial nidopallium (NCM), the songbird analog of the mammalian auditory association cortex. Multielectrode recordings coupled with local pharmacological manipulations in awake animals reveal that both exogenous and locally generated estradiol increase auditory-evoked activity in NCM. This enhancement in neuronal responses is mediated by suppression of local inhibitory transmission. Surprisingly, we also found that estradiol is both necessary and sufficient for the induction of multiple mitogen-activated protein kinase (MAPK)-dependent genes thought to be required for synaptic plasticity and memorization of birdsong. Specifically, we show that local blockade of estrogen receptors or aromatase activity in awake birds decrease song-induced MAPK-dependent gene expression. Infusions of estradiol in acoustically isolated birds induce transcriptional activation of these genes to levels comparable with song-stimulated animals. Our results reveal acute and rapid nongenomic functions for estradiol in central auditory physiology and suggest that such roles may be ubiquitously expressed across sensory systems.
14172390
Estradiol-dependent modulation of serotonergic markers in auditory areas of a seasonally breeding songbird.
Because no organism lives in an unchanging environment, sensory processes must remain plastic so that in any context, they emphasize the most relevant signals. As the behavioral relevance of sociosexual signals changes along with reproductive state, the perception of those signals is altered by reproductive hormones such as estradiol (E2). We showed previously that in white-throated sparrows, immediate early gene responses in the auditory pathway of females are selective for conspecific male song only when plasma E2 is elevated to breeding-typical levels. In this study, we looked for evidence that E2-dependent modulation of auditory responses is mediated by serotonergic systems. In female nonbreeding white-throated sparrows treated with E2, the density of fibers immunoreactive for serotonin transporter innervating the auditory midbrain and rostral auditory forebrain increased compared with controls. E2 treatment also increased the concentration of the serotonin metabolite 5-HIAA in the caudomedial mesopallium of the auditory forebrain. In a second experiment, females exposed to 30 min of conspecific male song had higher levels of 5-HIAA in the caudomedial nidopallium of the auditory forebrain than birds not exposed to song. Overall, we show that in this seasonal breeder, (a) serotonergic fibers innervate auditory areas; (b) the density of those fibers is higher in females with breeding-typical levels of E2 than in nonbreeding, untreated females; and (c) serotonin is released in the auditory forebrain within minutes in response to conspecific vocalizations. Our results are consistent with the hypothesis that E2 acts via serotonin systems to alter auditory processing.
14290383
Immediate early gene expression following exposure to acoustic and visual components of courtship in zebra finches
Sensory driven immediate early gene expression (IEG) has been a key tool to explore auditory perceptual areas in the avian brain. Most work on IEG expression in songbirds such as zebra finches has focused on playback of acoustic stimuli and its effect on auditory processing areas such as caudal medial mesopallium (CMM) caudal medial nidopallium (NCM). However, in a natural setting, the courtship displays of songbirds (including zebra finches) include visual as well as acoustic components. To determine whether the visual stimulus of a courting male modifies song-induced expression of the IEG ZENK in the auditory forebrain we exposed male and female zebra finches to acoustic (song) and visual (dancing) components of courtship. Birds were played digital movies with either combined audio and video, audio only, video only, or neither audio nor video (control). We found significantly increased levels of Zenk response in the auditory region CMM in the two treatment groups exposed to acoustic stimuli compared to the control group. The video only group had an intermediate response, suggesting potential effect of visual input on activity in these auditory brain regions. Finally, we unexpectedly found a lateralization of Zenk response that was independent of sex, brain region, or treatment condition, such that Zenk immunoreactivity was consistently higher in the left hemisphere than in the right and the majority of individual birds were left-hemisphere dominant.
30553052
Hemispheric specialization in the primary auditory area of awake and anesthetized starlings (Sturnus vulgaris).
Although evidence exists for a lateralization of song production, few studies have focused on the perceptual aspect of lateralization in songbirds. In the present study, the authors recorded neuronal responses to a variety of species-specific and artificial, nonspecific stimuli in both hemispheres of awake and anesthetized male starlings (Sturnus vulgaris). Recordings were made in the primary auditory area of the songbird brain, the Field L complex. The right hemisphere exhibited significantly more responsive units than the left hemisphere in awake birds, and this difference was significantly reduced in anesthetized birds. Furthermore, clear hemispheric specialization toward categories of behaviorally relevant stimuli and precise parameters of these stimuli were found. The main auditory area of the starling's brain thus appears to show some degree of lateralization.
1071442
Hemispheric differences in processing of vocalizations depend on early experience
An intriguing phenomenon in the neurobiology of language is lateralization: the dominant role of one hemisphere in a particular function. Lateralization is not exclusive to language because lateral differences are observed in other sensory modalities, behaviors, and animal species. Despite much scientific attention, the function of lateralization, its possible dependence on experience, and the functional implications of such dependence have yet to be clearly determined. We have explored the role of early experience in the development of lateralized sensory processing in the brain, using the songbird model of vocal learning. By controlling exposure to natural vocalizations (through isolation, song tutoring, and muting), we manipulated the postnatal auditory environment of developing zebra finches, and then assessed effects on hemispheric specialization for communication sounds in adulthood. Using bilateral multielectrode recordings from a forebrain auditory area known to selectively process species-specific vocalizations, we found that auditory responses to species-typical songs and long calls, in both male and female birds, were stronger in the right hemisphere than in the left, and that right-side responses adapted more rapidly to stimulus repetition. We describe specific instances, particularly in males, where these lateral differences show an influence of auditory experience with song and/or the bird’s own voice during development.
1753247
Own-Song Recognition in the Songbird Auditory Pathway: Selectivity and Lateralization
The songbird brain is able to discriminate between the bird's own song and other conspecific songs. Determining where in the brain own- song selectivity emerges is of great importance because experience-dependent mechanisms are necessarily involved and because brain regions sensitive to self-generated vocalizations could mediate auditory feedback that is necessary for song learning and maintenance. Using functional MRI, here we show that this selectivity is present at the midbrain level. Surprisingly, the selectivity was found to be lateralized toward the right side, a finding reminiscent of the potential right lateralization of song production in zebra finches but also of own-face and own-voice recognition in human beings. These results indicate that a midbrain structure can process subtle information about the identity of a subject through experience-dependent mechanisms, challenging the classical perception of subcortical regions as primitive and nonplastic structures. They also open questions about the evolution of the cognitive skills and lateralization in vertebrates.
27980289
Midbrain dopamine neurons reflect affiliation phenotypes in finches and are tightly coupled to courtship
Mesolimbic dopamine (DA) circuits mediate a wide range of goal-oriented behavioral processes, and DA strongly influences appetitive and consummatory aspects of male sexual behavior. In both birds and mammals, mesolimbic projections arise primarily from the ventral tegmental area (VTA), with a smaller contribution from the midbrain central gray (CG). Despite the well known importance of the VTA cell group for incentive motivation functions, relationships of VTA subpopulations to specific aspects of social phenotype remain wholly undescribed. We now show that in male zebra finches (Estrildidae: Taeniopygia guttata), Fos activity within a subpopulation of tyrosine hydroxylase-immunoreactive (TH-ir; presumably dopaminergic) neurons in the caudal VTA is significantly correlated with courtship singing and coupled to gonadal state. In addition, the number of TH-ir neurons in this caudal subpopulation dichotomously differentiates courting from non-courting male phenotypes, and evolves in relation to sociality (flocking vs. territorial) across several related finch species. Combined, these findings for the VTA suggest that divergent social phenotypes may arise due to the differential assignment of “incentive value” to conspecific stimuli. TH-ir neurons of the CG (a population of unknown function in mammals) exhibit properties that are even more selectively and tightly coupled to the expression of courtship phenotypes (and appetitive courtship singing), both in terms of TH-ir cell number, which correlates significantly with constitutive levels of courtship motivation, and with TH-Fos colocalization, which increases in direct proportion to the phasic expression of song. We propose that these neurons may be core components of social communication circuits across diverse vertebrate taxa.
27544006
Tyrosine hydroxylase and regulation of dopamine synthesis.
Tyrosine hydroxylase is the rate-limiting enzyme of catecholamine biosynthesis; it uses tetrahydrobiopterin and molecular oxygen to convert tyrosine to DOPA. Its amino terminal 150 amino acids comprise a domain whose structure is involved in regulating the enzyme's activity. Modes of regulation include phosphorylation by multiple kinases at four different serine residues, and dephosphorylation by two phosphatases. The enzyme is inhibited in feedback fashion by the catecholamine neurotransmitters. Dopamine binds to TyrH competitively with tetrahydrobiopterin, and interacts with the R domain. TyrH activity is modulated by protein-protein interactions with enzymes in the same pathway or the tetrahydrobiopterin pathway, structural proteins considered to be chaperones that mediate the neuron's oxidative state, and the protein that transfers dopamine into secretory vesicles. TyrH is modified in the presence of NO, resulting in nitration of tyrosine residues and the glutathionylation of cysteine residues.
37498699
Depolarisation‐Dependent Protein Phosphorylation and Dephosphorylation in Rat Cortical Synaptosomes Is Modulated by Calcium
Abstract: The effect of calcium on protein phosphorylation was investigated using intact synaptosomes isolated from rat cerebral cortex and prelabelled with 32Pi. For nondepolarised synaptosomes a group of calcium‐sensitive phosphoproteins were maximally labelled in the presence of 0.1 mM calcium. The phosphorylation of these proteins was slightly decreased in the presence of strontium and absent in the presence of barium, consistent with the decreased ability of these cations to activate calcium‐stimulated protein kinases. Addition of calcium alone to synaptosomes prelabelled in its absence increased phosphorylation of a number of proteins. On depolarisation in the presence of calcium certain of the calcium‐sensitive phosphoproteins were further increased in labelling above nondepolarised levels. These increases were maximal and most sustained after prelabelling at 0.1 mM calcium. On prolonged depolarisation at this calcium concentration a slow decrease in labelling was observed for most phosphoproteins, whereas a greater rate and extent of decrease occurred at higher calcium concentrations. At 2.5 mM calcium a rapid and then a subsequent slow dephosphorylation was observed, indicating two distinct phases of dephosphorylation. Of all the phosphoproteins normally stimulated by depolarisation, only phosphoprotein 59 did not exhibit the rapid phase of dephosphorylation at high calcium concentrations. Replacing calcium with strontium markedly decreased the extent of change observed on depolarisation whereas barium decreased phosphorylation changes even further. Taken together these data suggest that an influx of calcium into synaptosomes initially activates protein phosphorylation, but as the levels of intrasynaptosomal calcium rise protein dephosphorylation predominates. Other phosphoproteins were dephosphorylated immediately on depolarisation in the presence of calcium. The fine control of protein phosphorylation levels exerted by calcium supports the idea that the synaptosomal phosphoproteins could play a role in modulating events such as neurotransmitter release in the nerve terminal.
11995048
The effects of footshock and immobilization stress on tyrosine hydroxylase phosphorylation in the rat locus coeruleus and adrenal gland
Tyrosine hydroxylase (TH), the rate-limiting enzyme in catecholamine biosynthesis, is regulated acutely by protein phosphorylation. No studies have systematically investigated the time course of TH phosphorylation in vivo in response to different stressors. We therefore determined the extent of TH phosphorylation at Ser19, Ser31, and Ser40 over a 40-min period in response to footshock or immobilization stress in the rat locus coeruleus and adrenal medulla. There were significant changes in TH phosphorylation in both tissues and the responses to the two stressors differed markedly. With each of the phosphorylation sites immobilization stress caused a much smaller, or less sustained, response than footshock stress. With immobilization stress there was a transient increase in Ser31 phosphorylation in the locus coeruleus and in the adrenal medulla, but there were no effects on Ser19 or Ser40 phosphorylation. With footshock stress there was a substantial decrease in Ser19 phosphorylation over time, a substantial increase in Ser31 phosphorylation over time, but there were no effects on Ser40 phosphorylation. Measuring TH phosphorylation at Ser19, Ser31, and Ser40 over time can therefore be used as a sensitive index to differentiate the effects of different stressors on catecholaminergic cells.
41861990
Activation of tyrosine hydroxylase in the lateral efferent terminals by sound conditioning
Preconditioning to sound is a well-documented strategy to provide protections against a subsequent acoustic trauma. In the present study, preconditioning (1.0 kHz tone at 81 dB sound pressure level (SPL) for 24 h) protected ABR thresholds by 17-28 dB from an acoustic trauma (2.7 kHz, 103 dB SPL, 30 min) that resulted in a temporary threshold shift. The protection afforded by sound conditioning was shown to be blocked by the administration of 6-hydroxydopamine which disrupts tyrosine hydroxylase in the nerve terminals of the lateral efferent fibers. Furthermore, tyrosine hydroxylase immunoreactivity was up-regulated both by sound conditioning alone, and by the combined treatment of sound conditioning and acoustic trauma. In contrast, acoustic trauma alone resulted in a reduction in tyrosine hydroxylase immunoreactivity compared to unexposed controls. These findings are the first demonstration that tyrosine hydroxylase in the lateral efferents are up-regulated during sound conditioning and suggests a role for the lateral efferent system in protecting against acoustic trauma by sound conditioning.
32592537
Specific sound-induced noradrenergic and serotonergic activation in central auditory structures
We have studied the noradrenergic and serotonergic changes induced by white noise stimulation at 70, 90 or 110 dB SPL for 45 min, in cochlear nuclei, inferior colliculus (IC), primary auditory cortex (PAC) and as a comparison in locus coeruleus (LC) and raphe dorsalis using HPLC. Both noradrenergic and serotonergic pathways were activated in the dorsal+posteroventral cochlear nuclei (DCN+PVCN) without changes in the anteroventral cochlear nucleus (AVCN) and IC. In the DCN+PVCN the noradrenergic activation was restricted to animals exposed to 70 dB SPL whereas the increase of serotonin content was intensity-dependent. In PAC serotonergic activation was observed only after 70 dB SPL exposure. These data suggest that in physiological conditions (70 dB SPL) noradrenergic and serotonergic regulation of the processing of auditory information occurs specifically in the dorsal cochlear nucleus where the control of incoming information to higher auditory structures takes place (i.e. IC and PAC). We suggest that the serotonergic activation in the primary auditory cortex for 70 dB SPL sound stimulation could be related to the fact that low-intensity white noise stimulation could be the most plastic-demanding processing in the auditory cortex.
24776718
The Effect of Social Defeat on Tyrosine Hydroxylase Phosphorylation in the Rat Brain and Adrenal Gland
Tyrosine hydroxylase (TH), the rate-limiting enzyme in catecholamine biosynthesis, is regulated acutely by protein phosphorylation and chronically by protein synthesis. No studies have systematically investigated the phosphorylation of these sites in vivo in response to stressors. We specifically investigated the phosphorylation of TH occurring within the first 24 h in response to the social defeat stress in the rat adrenal, the locus coeruleus, substantia nigra and ventral tegmental area. Five groups were investigated; home cage control (HCC), two groups that underwent social defeat (SD+) which were sacrificed either 10 min or 24 h after the end of the protocol and two groups that were put into the cage without the resident being present (SD−) which were sacrificed at time points identical to the SD+. We found at 10 min there were significant increases in serine 40 and 31 phosphorylation levels in the locus coeruleus in SD+ compared to HCC and increases in serine 40 phosphorylation levels in the substantia nigra in SD+ compared to SD−. We found at 24 h there were significant increases in serine 19 phosphorylation levels in the ventral tegmental area in SD+ compared to HCC and decreases in serine 40 phosphorylation levels in the adrenal in SD+ compared to SD−. These findings suggest that the regulation of TH phosphorylation in different catecholamine-producing cells varies considerably and is dependent on both the nature of the stressor and the time at which the response is analysed.
26358104
Interstitial 3‐Methoxytyramine Reflects Striatal Dopamine Release: An In Vivo Microdialysis Study
Previous ex vivo studies have provided indirect evidence that the dopamine (DA) metabolite 3‐methoxytyramine (3‐MT) may be a useful index of DA release in vivo. In the present study, in vivo microdialysis was utilized to assess directly the relationship between extracellular DA and 3‐MT in the striatum of rats following a variety of pharmacological manipulations. Apomorphine, a DA receptor agonist, produced a rapid, transient decrease in both DA and 3‐MT. Conversely, the DA receptor antagonist haloperidol produced a concomitant increase in extracellular DA and 3‐MT. Increases in DA and 3‐MT were also noted following the administration of the DA uptake inhibitor, bupropion. Local application of tetrodotoxin resulted in the complete elimination of measurable amounts of DA and 3‐MT in the dialysate. γ‐Butyrolactone also greatly decreased DA and 3‐ MT. Finally, d‐amphetamine produced a large increase in DA and 3‐MT in animals that had been treated previously with γ‐butyrolactone. The Pearson correlation coefficients for DA and 3‐MT following these manipulations ranged from 0.87 to 0.97. These data indicate that interstitial 3‐MT is an accurate index of DA release. However, when compared with previous ex vivo findings, the present results also suggest that changes in tissue concentrations of 3‐MT may not reliably reflect DA release following certain pharmacological manipulations.
27419208
Drug‐Induced Changes in the Efflux of Dopamine and Serotonin Metabolites from the Brains of Freely Moving Rats a
A variety of techniques have been employed over the past 30 years to monitor the release of putative neurotransmitters from neurons in the central nervous system. The present report will be limited to studies on the release of serotonin (5-hydroxytryptamine, 5HT) and dopamine (DA) and their major metabolites (5-hydroxyindoleacetic acid, SHIAA; dihydroxyphenylacetic acid, DOPAC; homovanillic acid, HVA). These polar compounds do not readily pass the blood-brain barrier, but they do diffuse from the brain extracellular fluid into the cerebrospinal fluid (CSF). Accordingly, many investigators have analyzed for these compounds in CSF or in perfusates of the ventriculo-cisternal system. In the 1960s Feldberg and co-workers developed and employed a variety of techniques for use in anesthetized cats by which artificial CSF could be introduced through cannulae implanted into the lateral, third, or fourth ventricle and collected from cannulae or catheters inserted into any of these ventricles, the cerebral aqueduct, or the cisterna.' With these techniques it was possible to detect the efflux of acetylcholine, DA, HVA, 5HT, and 5HIAA?-*Subsequently, the Gaddum push-pull cannula: which is usually employed to perfuse discrete anatomical regions of brain tissue, was used to perfuse the lateral cerebral ventricles of unanesthetized rats.'o,'' Until recently most studies using cerebroventricular techniques in the rat have employed radioactive tracer techniques to quantify the release of DA, 5HT, and their metabolites."'12 The development of an analytical procedure that employs high performance liquid chromatography coupled to an electrochemical detection unit (LCEC) has made it possible to detect picogram quantities of these compounds in brain perfusates or CSF.'3s14 The present report will focus on results of studies in which this analytical technique has been employed to measure the concentrations of endogenous DOPAC and SHIAA in perfusates collected from push-pull cannulae that have been permanently implanted into a lateral cerebral ventricle of rats. The effects of phar-
39193110
Dopaminergic and Serotonergic Neurotransmission Systems Are Differentially Involved in Auditory Cortex Learning: A Long‐Term Microdialysis Study of Metabolites
Abstract: Auditory cortex has been shown to be a site of widespread neuronal learning processes even in the context of simple auditory conditioning behavior. In view of their presumed role in determining behavioral and motivational relevance of incoming information we investigated whether the dopaminergic and serotonergic systems are involved in auditory cortex learning. Using a chronic brain microdialysis technique over 4 days, samples from auditory cortex were obtained before, during, and after daily footshock avoidance training simultaneously from trained gerbils and passive control animals or pseudotrained animals. Because of detection limits of dopamine and serotonin in auditory cortex, the response profiles of extracellular homovanillic acid as the metabolite of the dopaminergic system and of 5‐hydroxyindoleacetic acid as the metabolite of the serotonergic system were determined from consecutive dialysis samples each day. The response of the dopaminergic system appeared to reflect the initial formation of the behaviorally relevant association exclusively during the first training day, whereas the serotonergic response appeared to correlate with the stress level of animals.
19861354
Does brain 3,4‐dihydroxyphenylacetic acid reflect dopamine release?
After sulpiride (75 mg kg−1) administration, a significant increase in dopamine, 3,4‐dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) occurred in the rat striatum. No attenuation in the sulpiride‐induced dopamine and DOPAC increase was seen in the striatum from rats previously treated with benztropine (25 mg kg−1). On the other hand, in the benztropine + sulpiride‐treated group, HVA levels were significantly lower compared with those of rats to which only sulpiride had been given. The results presented suggest that under appropriate conditions, as when sulpiride is used, dopamine can be deaminated before its release has occurred.
24188442
A kinetic study of the rate of formation of dopamine, 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) in the brain of the rat: Implications for the origin of dopac
The rates of disappearance of dopamine, 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) in the brain of rats treated with the tyrosine hydroxylase inhibitor alpha-methyl-p-tyrosine were measured. Levels of dopamine. DOPAC and HVA in tissue were quantified in the striatum, nucleus accumbens and frontal cortex of the rat by means of high pressure liquid chromatography with electrochemical detection. Rats were sacrificed 1, 2 or 3 hr after the administration of alpha-methyl-p-tyrosine (200 mg/kg, i.p.). Levels of dopamine, DOPAC and HVA in tissue were logarithmically transformed, plotted against the duration of inhibition of tyrosine hydroxylase and the rate constant of the decline of dopamine and of its metabolites calculated. The rate constant of decline of DOPAC in each of brain studied was found to be greater than that for dopamine. The rate constant for the decline of dopamine was found to be greater in areas of the brain presenting the largest DOPAC/dopamine tissue ratios (frontal cortex greater than nucleus accumbens greater than striatum). The present results suggest that a substantial amount of DOPAC in brain derives from a newly formed pool of dopamine.
4592617
In vivo measurement of extracellular dopamine and DOPAC in rat striatum after various dopamine-releasing drugs; implications for the origin of extracellular DOPAC.
In order to further examine the likely origin of the dopamine (DA) metabolite, 3,4-dihydroxyphenylacetic acid (DOPAC), certain drugs known to release DA from different intraneuronal pools were tested for their effects on extracellular striatal DA and DOPAC levels by means of brain microdialysis in the halothane-anaesthetized rat. Amphetamine (10(-6) and 10(-5) M), nomifensine (10(-5) M), potassium chloride (30 and 60 mM), methylphenidate (10(-5) and 10(-4) M) and tyramine (10(-5) M), when added to the perfusion medium and administered locally into the striatum via the dialysis membrane, increased the level of DA in striatal perfusates during the 20 min of application. In comparison, the level of DOPAC in the perfusates was decreased by both amphetamine (10(-5) M) and potassium chloride (60 mM), but was not significantly changed by nomifensine, methylphenidate or tyramine. The effect of amphetamine (10(-6) M) and nomifensine (10(-5) M) on DA and DOPAC levels was further studied by administering the drugs over a longer period of time (3 X 20 min). Although both of these treatments produced a similar increase of DA, only amphetamine reduced the levels of DOPAC. DA (10(-4) but not (10(-5) M) increased the levels of DOPAC but this effect was also seen in DA-denervated animals. These data indicate that when the DA nerve terminal is exposed to drugs which release newly synthesized DA, DOPAC declines possibly because intraneuronal monoamine oxidase is deprived of its main substrate. We suggest that these findings support the hypothesis that a major portion of the DA metabolite, DOPAC, is derived from an intraneuronal pool of newly synthesized DA.
11066242
Differential Effect of Stress on In Vivo Dopamine Release in Striatum, Nucleus Accumbens, and Medial Frontal Cortex
Abstract: Microdialysis was used to assess extracellular dopamine in striatum, nucleus accumbens, and medial frontal cortex of unanesthetized rats both under resting conditions and in response to intermittent tail‐shock stress. The dopamine metabolites 3,4‐dihydroxy‐phenylacetic acid and homovanillic acid also were measured. The resting extracellular concentration of dopamine was estimated to be approximately 10 nM in striatum, 11 nM in nucleus accumbens, and 3 nM in medial frontal cortex. In contrast, the resting extracellular levels of 3,4‐dihydroxyphenylacetic acid and homovanillic acid were in the low micromolar range. Intermittent tail‐shock stress increased extracellular dopamine relative to baseline by 25% in striatum, 39% in nucleus accumbens, and 95% in medial frontal cortex. 3,4‐Dihydroxyphenylacetic acid and homovanillic acid also were generally increased by stress, although there was a great deal of variability in these responses. These data provide direct in vivo evidence for the global activation of dopaminergic systems by stress and support the concept that there exist regional variations in the regulation of dopamine release.
25468295
Stress‐Induced Sensitization of Dopamine and Norepinephrine Efflux in Medial Prefrontal Cortex of the Rat
Abstract: We examined whether prior exposure to chronic cold (17–28 days, 5°C) alters basal or stress‐evoked (30‐min tail shock) catecholamine release in medial prefrontal cortex, nucleus accumbens, and striatum, using in vivo microdialysis. Basal norepinephrine (NE) concentrations in medial prefrontal cortex did not differ between chronically cold‐exposed rats and naive control rats (2.7 ± 0.3 vs. 2.5 ± 0.2 pg/20 µl, respectively). Basal dopamine (DA) efflux in any of the brain regions was not significantly different between chronically cold‐exposed rats and naive rats. However, a trend for lower basal DA efflux in the cold‐exposed relative to naive rats was observed in medial prefrontal cortex (1.5 ± 0.2 vs. 2.2 ± 0.3 pg/20 µl, respectively), nucleus accumbens (3.7 ± 0.8 vs. 5.4 ± 0.9 pg/20 µl, respectively), and striatum (4.4 ± 0.5 vs. 7.2 ± 1.5 pg/20 µl, respectively). In medial prefrontal cortex of rats previously exposed to cold, tail shock elicited a greater increase from baseline in both DA and NE efflux relative to that measured in naive rats (DA, 2.3 ± 0.3 vs. 1.2 ± 0.1 pg, respectively; NE, 3.8 ± 0.4 vs. 1.4 ± 0.2 pg, respectively). However, in nucleus accumbens or striatum of rats previously exposed to cold, the stress‐induced increase in DA efflux was not significantly different from that of naive rats (nucleus accumbens, 1.8 ± 0.7 vs. 1.5 ± 0.3 pg, respectively; striatum, 1.9 ± 0.4 vs. 2.6 ± 0.7 pg, respectively). Thus, both cortical NE projections and cortically projecting DA neurons sensitize after chronic exposure to cold. In contrast, subcortical DA projections do not sensitize under these conditions.
25238070
Predator stress engages corticotropin-releasing factor and opioid systems to alter the operating mode of locus coeruleus norepinephrine neurons
The norepinephrine nucleus, locus coeruleus (LC), has been implicated in cognitive aspects of the stress response, in part through its regulation by the stress-related neuropeptide, corticotropin-releasing factor (CRF). LC neurons discharge in tonic and phasic modes that differentially modulate attention and behavior. Here, the effects of exposure to an ethologically relevant stressor, predator odor, on spontaneous (tonic) and auditory-evoked (phasic) LC discharge were characterized in unanesthetized rats. Similar to the effects of CRF, stressor presentation increased tonic LC discharge and decreased phasic auditory-evoked discharge, thereby decreasing the signal-to-noise ratio of the sensory response. This stress-induced shift in LC discharge toward a high tonic mode was prevented by a CRF antagonist. Moreover, CRF antagonism during stress unmasked a large decrease in tonic discharge rate that was opioid mediated because it was prevented by pretreatment with the opiate antagonist, naloxone. Elimination of both CRF and opioid influences with an antagonist combination rendered LC activity unaffected by the stressor. These results demonstrate that both CRF and opioid afferents are engaged during stress to fine-tune LC activity. The predominant CRF influence shifts the operational mode of LC activity toward a high tonic state that is thought to facilitate behavioral flexibility and may be adaptive in coping with the stressor. Simultaneously, stress engages an opposing opioid influence that restrains the CRF influence and may facilitate recovery toward pre-stress levels of activity. Changes in the balance of CRF:opioid regulation of the LC could have consequences for stress vulnerability.
13069549
Context-dependent modulation of auditory processing by serotonin
Context-dependent plasticity in auditory processing is achieved in part by physiological mechanisms that link behavioral state to neural responses to sound. The neuromodulator serotonin has many characteristics suitable for such a role. Serotonergic neurons are extrinsic to the auditory system but send projections to most auditory regions. These projections release serotonin during particular behavioral contexts. Heightened levels of behavioral arousal and specific extrinsic events, including stressful or social events, increase serotonin availability in the auditory system. Although the release of serotonin is likely to be relatively diffuse, highly specific effects of serotonin on auditory neural circuitry are achieved through the localization of serotonergic projections, and through a large array of receptor types that are expressed by specific subsets of auditory neurons. Through this array, serotonin enacts plasticity in auditory processing in multiple ways. Serotonin changes the responses of auditory neurons to input through the alteration of intrinsic and synaptic properties, and alters both short- and long-term forms of plasticity. The infrastructure of the serotonergic system itself is also plastic, responding to age and cochlear trauma. These diverse findings support a view of serotonin as a widespread mechanism for behaviorally relevant plasticity in the regulation of auditory processing. This view also accommodates models of how the same regulatory mechanism can have pathological consequences for auditory processing.
14115610
Recent experience modulates forebrain gene–expression in response to mate–choice cues in European starlings
Mate–choice decisions can be experience dependent, but we know little about how the brain processes stimuli that release such decisions. Female European starlings (Sturnus vulgaris) prefer males with long–bout songs over males with short–bout songs, and show higher expression of the immediate early gene (IEG) ZENK in the auditory forebrain when exposed to long–bout songs than when exposed to short–bout songs. We exposed female starlings to a short–day photoperiod for one of three durations and then, on an increased photophase, exposed them to one week of long–bout or short–bout song experience. We then examined their IEG response to novel long–bout versus novel short–bout songs by quantifying ZENK protein in two song–processing areas: the caudo–medial hyperstriatum ventrale and the caudo–medial neostriatum. ZENK expression in both areas increased with tenure on short–day photoperiods, suggesting that short days sensitize females to song. The ZENK response bias toward long–bout songs was greater in females with long–bout experience than in females with short–bout experience, indicating that the forebrain response bias toward a preferred trait depends on recent experience with that category of trait. This surprising level of neuroplasticity is immediately relevant to the natural history and fitness of the organism, and may underlie a mechanism for optimizing mate–choice criteria amidst locally variable distributions of secondary sexual characteristics.
27276632
Estrogen‐dependent selectivity of genomic responses to birdsong
Behavioral responses to sociosexual signals often depend on gonadal steroid hormones, which are thought to modulate behavior by acting on motivational systems in the brain. There is mounting evidence that sex steroids may also modulate perception of sociosexual signals by affecting sensory processing. In seasonally breeding songbirds such as the white‐throated sparrow (Zonotrichia albicollis), the female's behavioral response to hearing male song depends on her plasma levels of estradiol (E2). Here, we examined whether plasma E2 also affects the selectivity of the song‐induced zenk (egr‐1) response in the auditory forebrain, which is known to vary according to the behavioral relevance of song stimuli. Non‐breeding females were held on a winter‐like photoperiod and implanted with silastic capsules containing either no hormone or E2. E2‐treated birds hearing 42 min of conspecific song had more cells immunoreactive for the protein product of zenk in the auditory forebrain than did those hearing frequency‐matched synthetic tones. In birds not treated with E2, however, the zenk response to song did not differ from that to tones. We found similar effects in the avian homolog of the inferior colliculus, indicating that E2 may affect the processing of auditory information upstream of the forebrain. Our data suggest that in females, zenk induction in the auditory system is selective for song only when plasma E2 exceeds non‐breeding levels. E2‐dependent plasticity of auditory pathways and processing centres may promote recognition of and attention to conspecific song during the breeding season.
43759679
Topography of estradiol‐modulated genomic responses in the songbird auditory forebrain
Sex steroids facilitate dramatic changes in behavioral responses to sociosexual signals and are increasingly implicated in the sensory processing of those signals. Our previous work demonstrated that in female white‐throated sparrows, which are seasonal breeders, genomic responses in the auditory forebrain are selective for conspecific song over frequency‐matched tones only when plasma estradiol (E2) reaches breeding levels. Here, we sought to map this E2‐dependent selectivity in the best‐studied area of the auditory forebrain, the caudomedial nidopallium (NCM). Nonbreeding females with low endogenous levels of E2 were treated with E2 or a placebo and exposed to conspecific song, tones, or no sound playback. Immunoreactive protein product of the immediate early gene zenk (egr‐1) was then quantified within seven distinct subregions, or domains, of NCM. We report three main findings: (1) regardless of hormone treatment, the zenk response is significantly higher in dorsal than in ventral NCM, and higher in medial than in lateral NCM; (2) E2‐dependent selectivity of the response is limited to the rostral and medial domains of NCM; in the more caudal domains, song induces more zenk expression than tones regardless of hormone treatment; (3) even when no sound stimuli were presented, E2 treatment significantly increased zenk expression in the rostral, but not the caudal, domains of NCM. Together, the latter two findings suggest that E2‐dependent plasticity in NCM is concentrated in rostral NCM, which is hodologically and neurochemically distinct from caudal NCM. Activity in rostral NCM may therefore be seasonally regulated in this species. © 2009 Wiley Periodicals, Inc. Develop Neurobiol 2010
23433531
Dissecting natural sensory plasticity: Hormones and experience in a maternal context
There is a growing consensus that the auditory system is dynamic in its representation of behaviorally relevant sounds. The auditory cortex in particular seems to be an important locus for plasticity that may reflect the memory of such sounds, or functionally improve their processing. The mechanisms that underlie these changes may be either intrinsic because they depend on the receiver's physiological state, or extrinsic because they arise from the context in which behavioral relevance is gained. Research in a mouse model of acoustic communication between offspring and adult females offers the opportunity to explore both of these contributions to auditory cortical plasticity in a natural context. Recent works have found that after the vocalizations of infant mice become behaviorally relevant to mothers, auditory cortical activity is significantly changed in a way that may improve their processing. Here we consider the hypothesis that maternal hormones (intrinsic factor) and sensory experience (extrinsic factor) contribute together to drive these changes, focusing specifically on the evidence that well-known experience-dependent mechanisms of cortical plasticity can be modulated by hormones.
16963081
A DNA test to sex most birds
Birds are difficult to sex. Nestlings rarely show sex‐linked morphology and we estimate that adult females appear identical to males in over 50% of the world's bird species. This problem can hinder both evolutionary studies and human‐assisted breeding of birds. DNA‐based sex identification provides a solution. We describe a test based on two conserved CHD (chromo‐helicase‐DNA‐binding) genes that are located on the avian sex chromosomes of all birds, with the possible exception of the ratites (ostriches, etc.; Struthioniformes). The CHD‐W gene is located on the W chromosome; therefore it is unique to females. The other gene, CHD‐Z, is found on the Z chromosome and therefore occurs in both sexes (female, ZW; male, ZZ). The test employs PCR with a single set of primers. It amplifies homologous sections of both genes and incorporates introns whose lengths usually differ. When examined on a gel there is a single CHD‐Z band in males but females have a second, distinctive CHD‐W band.
2713755
THE NATURAL TERMINATION OF THE REFRACTORY PERIOD IN THE SLATE-COLORED JUNCO AND IN THE WHITE-THROATED SPARROW
It has been demonstrated that birds of several passefine species show gonadal recrudescence within six or seven weeks, when given sufficient artificial increases in day length in late fall or in winter. Birds treated in late summer or early fall, however, do not respond. The period between regression of the gonad, normally at the end of the breeding season, and the time the gonad responds to photostimulation, has been designated the refractory period. The experiments reported here were undertaken to determine by systematic study when the refractory period ends in the Slate-colored Junco (]unco hyemalis) and in the White-throated Sparrow (Zonotrichia albicollis)--species in which the natural termination of the refractory period had not been investigated. Secondarily, the effect of treatment with short days in the fall on the duration of the refractory period in these species was studied. It is with a great deal of pleasure that I acknowledge the assistance and helpful criticism of Dr. Albert Wolfson. The research reported here was supported in part by a grant from the Graduate School of Northwestern University.
21275391
Activity of the hypothalamic-pituitary-gonadal axis differs between behavioral phenotypes in female white-throated sparrows (Zonotrichia albicollis).
The white-throated sparrow (Zonotrichia albicollis) lends itself particularly well to investigations of neuroendocrine mechanisms of social behavior because of a behavioral polymorphism that correlates with a plumage phenotype. Roughly half of the individuals of this species exhibit a white stripe (WS) on the crown and engage in a more aggressive strategy, whereas the other half exhibit a tan stripe (TS) and assume a more parental strategy. These behavioral differences are mirrored by hormonal and neuroendocrine differences; for example, males of the WS morph have higher plasma testosterone than do TS males, and females of the TS morph have higher plasma luteinizing hormone than females of the WS morph. These differences suggest that the regulation of the hypothalamic-pituitary-gonadal (HPG) axis may differ according to morph. In this study, we compared HPG axis activity at each level by measuring (1) the number, size, and staining intensity of GnRH immunoreactive (ir) neurons; (2) plasma LH; and (3) plasma estradiol (E2) in females. We found that TS females had more GnRH-ir neurons in the septo-preoptic area of the hypothalamus than did WS females, and GnRH-ir neuronal cell bodies were larger in the WS than the TS females. There was no morph difference in the intensity of GnRH labeling. TS females had higher plasma LH, which is consistent with a previous report, and higher plasma E2. We hypothesize that the differences in GnRH-ir cell number and size are related to differences in LH and E2 secretion, and may be relevant to polymorphic social behavior.
22914198
Rapid neuroendocrine responses to auditory courtship signals.
In many species, courtship signals enhance reproductive function in the receiver. How these social signals are processed by the brain, particularly how they induce an endocrine response, is not well understood. Songbirds provide an ideal model in which to study this phenomenon because of the large existing literature on both their auditory neurobiology and the control of their reproductive physiology by environmental cues. To date, all of the relevant studies on songbirds have involved measuring the effects of male vocalizations on ovarian function over a period of weeks, a time course that precludes detailed analysis of the neuroendocrine mechanisms operating during song perception. We played recordings of conspecific male song to laboratory-housed female white-throated sparrows and quantified the resulting rapid changes in LH as well as the induction of the immediate early gene Egr-1 in the GnRH system and mediobasal hypothalamus (MBH). Hearing song for 42 min induced LH release and Egr-1 expression in the MBH, but did not alter Egr-1 expression in GnRH neurons. The time course of LH release and the pattern of Egr-1 expression together suggest that song acts as a trigger to induce GnRH release in a manner resembling photostimulation. The Egr-1 response in the MBH was qualitatively distinguishable from the responses to either photostimulation or pharmacologically induced LH release but seemed to involve overlapping neuronal populations. Song-induced Egr-1 expression in the MBH was correlated with the expression in midbrain and forebrain auditory centers, further supporting a role for the MBH in processing social information.
85093486
Effect of female sexual displays on the endocrine physiology and behaviour of male white‐crowned sparrows, Zonotrichia leucophrys
In photoperiodic birds, endocrine responses to behavioural interactions between males and females may be involved in temporally “fine-tuning” the onset of reproduction to yearly variations in the environment. This study examined the endocrine and behavioural responses of male White-crowned sparrows (Zonotrichia leucophrys) to changes in the endocrine state of the female, as signalled by changes in her behaviour. Males on different photoperiodic regimes were paired with oestrogen-treated, sexually receptive females. Males exposed to gonadostimulatory long days mounted and copulated with oestrogen-treated females even before gonadal development was complete. These males had higher plasma levels of testosterone and luteinizing hormone and maintained enlarged testes longer than control males paired with untreated, nonreceptive females. Males maintained on nonstimulatory short days also mounted oestrogen-treated females; however, testes of these males remained nonfunctional and their plasma levels of testosterone and luteinizing hormone were basal. Thus, reproductive function of photostimulated males is profoundly affected by changes in the endocrine state and behaviour of the female. However, male sexual behaviours are expressed in response to visual and auditory stimuli from the female regardless of male hormonal condition or photoperiodic treatment.
5487355
Estradiol modulates neural responses to song in a seasonal songbird
Social behaviors such as courtship, parenting, and aggression depend primarily on two factors: a social signal to trigger the behavior, and a hormonal milieu that facilitates or permits it. Gonadal steroids may alter the valence or perceived context of the signal so that the same pheromone, vocalization, or visual display may elicit very different responses depending on the receiver's plasma hormone level. The neural processes underlying this phenomenon, however, are not well understood. Here, we describe how hormones modulate neural responses to social signals in female white‐throated sparrows listening to recordings of male song. While manipulating levels of the ovarian steroid estradiol, we mapped and quantified sound‐induced expression of the immediate early gene egr‐1 in nine brain regions that constitute a social behavior network in vertebrates. In most regions of interest, hearing male song induced more expression than hearing tones or silence, and this selectivity for song was seen only in birds with estradiol levels typical of the breeding season. In females with regressed ovaries and no exogenous estradiol, neural responses were selective for song over tones only in the lateral portion of the ventromedial hypothalamus, not in the rest of the network. Because the effects of hormone treatment on neural responses are not identical in each region, the overall pattern of activation across the network changes with estradiol level and thus with season and breeding context. Our results demonstrate a possible mechanism by which gonadal steroids may alter the processing of social signals and affect social decision‐making. J. Comp. Neurol. 511:173–186, 2008. © 2008 Wiley‐Liss, Inc.
35756401
Immediate early gene response to hearing song correlates with receptive behavior and depends on dialect in a female songbird
Stimulus-induced expression of the immediate early gene ZENK (egr-1) in the songbird's auditory forebrain presumably depends on the behavioral significance of the stimulus. Few studies, however, have quantified both the ZENK and behavioral responses to a stimulus in the same individuals. We played conspecific male song of either hatch (local) or foreign dialect to female white-crowned sparrows (Zonotrichia leucophrys oriantha) and quantified both the auditory ZENK response and their behavioral response, which is known to depend on dialect. Birds hearing hatch dialect showed greater ZENK induction in the caudomedial hyperstriatum ventrale and the dorsal portion of the caudomedial neostriatum than birds hearing foreign dialect, supporting previous work showing a relationship between ZENK and salience of the stimulus. In the dorsal portion of the caudomedial neostriatum, ZENK induction was correlated with the amount of non-vocal courtship behavior; however, in the caudomedial hyperstriatum ventrale, ZENK induction was more highly correlated with the females' own vocal behavior and thus may have been partly self-induced. Some females sang and showed a male-like pattern of ZENK induction in their song systems. This study provides the first evidence that the ZENK response in a sensory area to a social stimulus is proportional to the animal's preference for the stimulus.
23085836
Behavioral phenotypes persist after gonadal steroid manipulation in white-throated sparrows
White-throated sparrows (Zonotrichia albicollis) exhibit a behavioral polymorphism that segregates with a plumage marker. Individuals with a white stripe (WS) on the crown engage in an aggressive strategy that involves more singing, whereas individuals with a tan stripe (TS) sing less and engage in more parental care. Previous work has shown that plasma levels of gonadal steroids differ between the morphs in both sexes, suggesting a hormonal mechanism for the polymorphic behavior in this species. Here, we eliminated morph differences in plasma levels of testosterone (T) in males and estradiol (E2) in females in order to test whether morph differences in behavior would be similarly eliminated. Males and females in non-breeding condition were treated with T or E2, respectively, so that plasma levels in the treated groups were high and equal between the WS and TS morphs. We found that despite hormone treatment, WS and TS birds differed with respect to singing behavior. WS males sang more in response to song playback than did TS males, and WS females exhibited more spontaneous song than TS females. We also found that WS males gave more chip calls, which are often used in contexts of territorial aggression. Overall, these results suggest that WS birds engage in more territorial vocalization, particularly song, than do TS birds, even when T or E2 levels are experimentally equalized. This behavioral difference may therefore be driven by other factors, such as steroid metabolism, receptor expression or function, or steroid-independent neurotransmitter systems.
21447854
Neuroendocrine correlates of behavioral polymorphism in white-throated sparrows
Interspecific differences in the neuropeptide systems of the lateral septum (LS) often parallel differences in social behavior. In rodents, some closely related species that differ in aggressive behavior also differ according to the level of vasopressin (VP) innervation of the LS. In songbirds, the neuropeptides vasotocin (VT) and vasoactive intestinal peptide (VIP) affect aggression when administered directly to the LS. Here, we tested whether the density of VT or VIP innervation of the LS reflects patterns of intraspecific behavioral polymorphism in male and female white-throated sparrows (Zonotrichia albicollis), in which the "white-stripe" (WS) morph behaves more aggressively than the "tan-stripe" (TS) morph. We found that the WS birds had more VT-immunoreactivity (IR) than the TS birds in the ventrolateral subdivision of the caudal LS (LSc.vl) and in the medial portion of the bed nucleus of the stria terminalis (BSTm). In addition, the TS birds had more densely stained VIP-IR in the LSc.vl than the WS birds. Males had more VT-IR than females in the LSc.vl and BSTm, and more VIP-IR in the LSc.vl. We also report sex and morph differences in VIP-IR in the basal hypothalamus, where VIP is synthesized and released into the portal vasculature. Males had nearly twice as many VIP-immunoreactive (ir) neurons in the infundibular nucleus than did females, and birds of the WS morph had more densely stained VIP-IR in the median eminence than TS birds. Our results support the hypothesis that differences in these neuropeptide systems underlie inter- and intraspecific differences in social behavior across vertebrates.
22277929
The glucose oxidase-DAB-nickel method in peroxidase histochemistry of the nervous system
A combination of the glucose oxidase-diaminobenzidine (DAB) method and the DAB-nickel method can successfully bring out details of immunoreactive structures in immunostained preparations. It is especially beneficial for visualizing fibers and terminals.
34095330
Gonadal steroid receptor mRNA in catecholaminergic nuclei of the canary brainstem
Steroid actions in the song system may be modulated by ascending inputs from catecholaminergic (CA) brain nuclei; however, whether these nuclei contain steroid receptors is unknown. Here, we compared the distribution of androgen receptor (AR) and estrogen receptor-alpha (ER-alpha) mRNA with that of tyrosine hydroxylase immunoreactivity (TH-IR) in the brainstems of male canaries. Areas containing AR and ER-alpha mRNA overlapped with areas containing TH-IR cell bodies in the locus ceruleus and the area ventralis of Tsai. The substantia nigra and the midbrain central gray contained both TH-IR and AR mRNA. The presence of AR and ER-alpha within CA cell groups suggests that sex steroid hormones may modulate song production at the site of CA synthesis.
38330295
Magic peptides, magic antibodies: Guidelines for appropriate controls for immunohistochemistry
Since the earliest days of immunohistochemistry, the field has been troubled by the tendency for these methods to give spurious results. There are a number of reasons why various antibody preparations may provide variable and even erroneous results, and a series of steps one can take to control for this. Unfortunately, these are not as widely known as we feel they should be, even as we enter the fourth decade in which these methods have been widely available to neuroanatomists. Hence, we provide this editorial to help guide aspiring authors through the rocky shoals of immunolocalization in the nervous system.
5337879
Development of enteric and vagal innervation of the zebrafish (Danio rerio) gut
The autonomic nervous system develops following migration and differentiation of precursor cells originating in the neural crest. Using immunohistochemistry on intact zebrafish embryos and larvae we followed the development of the intrinsic enteric and extrinsic vagal innervation of the gut. At 3 days postfertilization (dpf), enteric nerve cell bodies and fibers were seen mainly in the middle and distal intestine, while the innervation of the proximal intestine was scarcer. The number of fibers and cell bodies gradually increased, although a large intraindividual variation was seen in the timing (but not the order) of development. At 11–13 dpf most of the proximal intestine received a similar degree of innervation as the rest of the gut. The main intestinal branches of the vagus were similarly often already well developed at 3 dpf, entering the gut at the transition between the proximal and middle intestine and projecting posteriorly along the length of the gut. Subsequently, fibers branching off the vagus innervated all regions of the gut. The presence of several putative enteric neurotransmitters was suggested by using markers for neurokinin A (NKA), pituitary adenylate cyclase‐activating polypeptide (PACAP), vasoactive intestinal polypeptide (VIP), nitric oxide, serotonin (5‐hydroxytryptamine, 5‐HT), and calcitonin gene‐related peptide (CGRP). The present results corroborate the belief that the enteric innervation is well developed before the onset of feeding (normally occurring around 5–6 dpf). Further, the more detailed picture of how development proceeds at stages previously not examined suggests a correlation between increasing innervation and more regular and elaborated motility patterns. J. Comp. Neurol. 508:756–770, 2008. © 2008 Wiley‐Liss, Inc.
30752400
The catecholaminergic system of the quail brain: Immunocytochemical studies of dopamine β‐hydroxylase and tyrosine hydroxylase
The distribution of dopamine β‐hydroxylase and tryrosine hydroxylase, two key enzymes in the biosynthesis of catecholamine, was investigated by immunocytochemistry in the brain of male and female Japanese quail. Cells or fibers showing dopamine β‐hydroxylase and tyrosine hydroxylase immunoreactivity were considered to be noradrenergic or adrenergic, while all structures showing only tyrosine hydroxylase immunoreactivity were tentatively considered to be dopaminergic. The major dopaminergic and noradrenergic cell groups that have been identified in the brain of mammals could be observed in the Japanese quail, with the exception of a tuberoinfundibular dopaminergic group. The dopamine β‐hydroxylase‐immunoreactive cells were found exclusively in the pons (locus ceruleus and nucleus subceruleus ventralis) and in the medulla (area of the nucleus reticularis). The tyrosine hydroxylase‐immunoreactive cells had a much wider distribution and extended from the preoptic area to the level of the medulla. They were, however, present in larger numbers in the area ventralis of Tsai and in the nucleus tegmenti pedunculo‐pontinus, pars compacta, which respectively correspond to the ventral tegmental area and to the substantia nigra of mammals. A high density of dopamine β‐hydroxylase‐ and tyrosine hydroxylase‐immunoreactive fibers and punctate structures was found in several steroid‐sensitive brain regions that are implicated in the control of reproduction. In the preoptic area and in the region of the nucleus accumbens‐nucleus stria terminalis, immunonegative perikarya were completely surrounded by immunoreactive fiber forming basket‐like structures. Given that some of these cells contain the enzyme aromatase, these structures may represent the morphological substrate for a regulation of aromatase activity by catecholamines. The dopamine β‐hydroxylase‐immunoreactive fibers were also present in a larger part of the preoptic area of females than in males. This sex difference in the noradrenergic innervation of the preoptic area presumably reflects the sex difference in norepinephrine content in this region. © 1993 Wiley‐Liss, Inc.
38480987
Development of the catecholaminergic innervation of the song system of the male zebra finch.
Catecholamines (CA) have been proposed to have neuromodulatory actions, particularly on attention and learning, in a number of neural systems. Because several of the interconnected brain nuclei that mediate song learning and production in the adult male zebra finch (Taeniopygia guttata) contain these neurotransmitters, we investigated the appearance of the catecholaminergic innervation of the song nuclei of male zebra finches during posthatch development, specifically during the period in which song learning occurs. We studied the development of immunoreactivity for tyrosine hydroxylase (TH) in the song nuclei HVc, RA, NIf, LMAN, and Area X in young males aged 20, 35, and 60 days as well as in adults (> 90 days). We also visualized catecholamines directly in Area X using CA histofluorescence. Both TH immunoreactivity and CA histofluorescence were initially low in Area X relative to their levels in the surrounding parolfactory lobe (LPO), and then increased during development to become more intense than in LPO by days 60-90. Similarly, TH immunoreactivity in HVc was initially low relative to that in the surrounding neostriatum, then increased during development to become more intense than that in the surround by day 60. TH immunostaining also increased markedly in NIf, RA, and LMAN over the same period. These results show that the levels of catecholamines and their major synthetic enzyme increase in song nuclei during development and thus raise the possibility that these transmitters contribute to the development of the song system or to song learning.
24218152
Distribution of tyrosine hydroxylase‐containing neurons and fibers in the brain of the budgerigar (Melopsittacus undulatus): General patterns and labeling in vocal control nuclei
The distribution of tyrosine hydroxylase (TH) was mapped out in cells and fibers of the budgerigar (Melopsittacus undulatus) brain. Special attention was given to vocal control and auditory nuclei because budgerigars are a psittacine species in which both males and females are capable of lifelong vocal learning (Farabaugh et al. [1994] J. Comp. Psychol 108:81–92). The results show that TH staining in the central nucleus of the anterior archistriatum (AAc) resembled that of surrounding archistriatal fields, except for portions of the ventral archistriatum, which exhibited substantially more TH+ fibers. Fewer fibers and fiber baskets are present in the central nucleus of the lateral neostriatum (NLc) than in surrounding fields. Both the oval nuclei of the ventral hyperstriatum (HVo) and anterior neostriatum (NAo) exhibit less fiber staining than surrounding fields whereas fiber staining in the medial NAo (NAom) and magnicellular nucleus of the parolfactory lobe (LPOm) resemble that of surrounding fields. Staining in primary telencephalic auditory nuclei was extremely low. The only sex difference observed was slightly increased TH staining in LPOm of females compared with surrounding fields on some tissue sections. These findings are in contrast to previous findings in zebra finch (Poephila guttata), a close ended vocal learning songbird in which TH staining in vocal nuclei increases during development and remains greater than surrounding fields throughout adulthood. The present results therefore support the view that catecholamines act to inhibit vocal plasticity in adult vocal learning species. Several unique features of TH‐immunoreactive (ir) cell groups were observed in the brainstem including sparsely scattered TH‐ir somata immediately adjacent to the third ventricle, within the tectum, basal forebrain, archistriatum, and caudal neostriatum, and in the hippocampus. These latter populations have not been described in other avian species and resemble features of the catecholamine system generally found in either reptiles or mammals. J. Comp. Neurol. 429:436–454, 2001. © 2000 Wiley‐Liss, Inc.
6720409
The distribution of tyrosine hydroxylase immunoreactivity in the brains of male and female zebra finches.
A system of brain nuclei controls song learning and behavior in zebra finches (Poephila guttata). The size of song-control nuclei are much larger in males, which sing, than in females, which do not sing. This study examined the distribution of fibers, terminals, and cell bodies that are immunoreactive for tyrosine hydroxylase (TH) (the rate-limiting enzyme in the synthesis of catecholamines) in song-control nuclei of adult males and females and juvenile males. In addition, the broad pattern of TH staining throughout the brain was described. There was a sex difference in TH immunoreactivity within song-control nuclei: males had light to moderate staining in all three cortical nuclei examined, whereas females had little or no label in corresponding areas [lateral magnocellular nucleus of the anterior neostriatum (IMAN), higher vocal center (HVC), and robust nucleus of the archistriatum (RA)]. The song-control nucleus area X (X), located in the striatum of avian basal ganglia, was more darkly stained than the surrounding striatum only in males; X was not defined by more intense immunoreactivity in females and hence could not be visualized. There were no apparent differences in TH staining in males ranging in age from 50 days to adulthood (> 90 days). Outside of the song-control system there were no substantive differences as a function of sex or age in the pattern or intensity of TH labeling. Major areas of telencephalic staining included the striatal region of basal ganglia, which was covered with dense, fine-grained label, and the septum, where cell bodies were encircled by extremely well-labeled thick processes. In the diencephalon, the preoptic area and hypothalamus included a complex pattern of darkly stained somata and fiber and terminal labeling. Darkly stained somata surrounded the pretectal nucleus, and labeled processes ramified throughout the superficial layers of the optic tectum. The midbrain and hindbrain contained a dense plexus of extremely dark cell bodies corresponding to mammalian substantia nigra, adjacent tegmental areas, and locus ceruleus. Labeled hindbrain cells were also seen in the pontine region, around nucleus solitarius, and in the ventrolateral medulla.
22289390
Organization of the dopaminergic innervation of forebrain areas relevant to learning: A combined immunohistochemical/retrograde tracing study in the domestic chick
The mediorostral neostriatum/hyperstriatum ventrale (MNH) and neostriatum dorsocaudale (Ndc) of the domestic chick are crucially involved in auditory filial imprinting, whereas the lobus parolfactorius (LPO) seems to be involved in the emotional modulation of behavior. Because there is evidence that MNH and Ndc are akin to higher association areas in mammals, the present study evaluates the dopaminergic and thalamic input to these areas, as well as to the avian caudate/putamen homologue LPO, by using retrograde pathway tracing, together with dopamine (DA) and tyrosine hydroxylase (TH) immunohistochemistry. By combining DA immunohistochemistry with retrograde fluorescent tracing, we demonstrated that dopaminergic afferents to the MNH and Ndc arise mainly from the area ventralis, whereas the main dopaminergic input to the LPO arises from the substantia nigra. The main thalamic input to the MNH and LPO arises from the dorsal thalamic nuclei, n. dorsomedialis anterior and n. dorsolateralis anterior, whereas the thalamic input to the Ndc arises from the n. dorsolateralis posterior and n. subrotundus. Furthermore, there are reciprocal intratelencephalic connections between distinct parts of the neostriatum caudale and the mediorostral neostriatum. DA‐immunoreactive (ir) fibers are present at moderate densities in the MNH and Ndc and at high densities in the LPO. At the ultrastructural level, DA‐ and TH‐ir axon terminals in the MNH and Ndc form predominantly symmetric synaptic contacts with dendritic shafts, which are often situated in close vicinity to unstained terminals. These results indicate that the general organization of dopaminergic afferents to the chick telecephalon is similar to that of the mesotelencephalic dopaminergic subsystems in mammals such as the mesostriatal and mesolimbocortical DA system. © 1996 Wiley‐Liss, Inc.
5891307
Distinction of Neurochemistry between the Cores and Their Shells of Auditory Nuclei in Tetrapod Species
The distribution of Met-enkephalin (ENK), substance P (SP) and serotonin (5-HT) differs between the core and shell regions of the mesencephalic and diencephalic auditory nuclei of the turtle [Belekhova et al., 2002]. These neurochemical distinctions are also found in other tetrapods (mammals, birds and amphibians). The distribution of ENK, SP and 5-HT was examined in the core and shell regions of both mesencephalic and diencephalic auditory nuclei, and in the telencephalic auditory areas of Bengalese finches (Lonchura striata) and mice (Mus musculus), as well as in corresponding auditory areas in toads (Bufo bufo). ENK, SP and 5-HT immunoreactive fibers and perikarya were largely absent from the core regions of both mesencephalic and diencephalic auditory nuclei, in comparison with the shell regions of mice and Bengalese finches. In the toad, however, this pattern was observed in the mesencephalic auditory nucleus, but not in the diencephalic auditory areas. ENK and SP immunoreactive perikarya were detected in the telencephalic auditory area of mice, whereas no ENK, SP or 5-HT immunolabeling was observed in the telencephalic auditory area (Field L) of Bengalese finches. These findings are discussed in terms of the evolution of the core-and-shell organization of auditory nuclei of tetrapods.
13320741
Subdivisions of the Auditory Midbrain (N. Mesencephalicus Lateralis, pars dorsalis) in Zebra Finches Using Calcium-Binding Protein Immunocytochemistry
The midbrain nucleus mesencephalicus lateralis pars dorsalis (MLd) is thought to be the avian homologue of the central nucleus of the mammalian inferior colliculus. As such, it is a major relay in the ascending auditory pathway of all birds and in songbirds mediates the auditory feedback necessary for the learning and maintenance of song. To clarify the organization of MLd, we applied three calcium binding protein antibodies to tissue sections from the brains of adult male and female zebra finches. The staining patterns resulting from the application of parvalbumin, calbindin and calretinin antibodies differed from each other and in different parts of the nucleus. Parvalbumin-like immunoreactivity was distributed throughout the whole nucleus, as defined by the totality of the terminations of brainstem auditory afferents; in other words parvalbumin-like immunoreactivity defines the boundaries of MLd. Staining patterns of parvalbumin, calbindin and calretinin defined two regions of MLd: inner (MLd.I) and outer (MLd.O). MLd.O largely surrounds MLd.I and is distinct from the surrounding intercollicular nucleus. Unlike the case in some non-songbirds, however, the two MLd regions do not correspond to the terminal zones of the projections of the brainstem auditory nuclei angularis and laminaris, which have been found to overlap substantially throughout the nucleus in zebra finches.
24454165
Simultaneous quantification of dopamine, 5-hydroxytryptamine and four metabolically related compounds by means of reversed-phase high-performance liquid chromatography with electrochemical detection.
A method for simultaneously quantifying dopamine, 5-hydroxytryptamine (5-HT) and four metabolically related compounds has been developed, permitting more efficient neurochemical examination of these often interrelated biogenic amine systems. The method uses high-performance liquid chromatographic separation of these compounds on a C18 reversed-phase column with a buffered mobile phase containing methanol as an organic modifier and heptanesulfonate as an ion-pair reagent. Using 5-hydroxy-N-methyltryptamine as an internal standard and electrochemical detection, chromatography time is less than 12 min. Sample preparation simply involves the addition of internal standard, homogenization in the mobile phase, centrifugation and injection of the supernatant into the chromatograph. The method is sensitive to a tissue content of these compounds of less than 1 ng. The utility of this method for neuropharmacological--neurochemical studies is illustrated with studies using inhibitors of monoamine oxidase (pargyline) and aromatic amino acid decarboxylase (RO 4-4602).