182952126
Neuronal cell-subtype specificity of neural synchronization in mouse primary visual cortex
Spatiotemporally synchronised neuronal activity is central to sensation, motion and cognition. Brain circuits consist of dynamically interconnected neuronal cell-types, thus elucidating how neuron types synergise within the network is key to understand the neuronal orchestra. Here we show that in neocortex neuron-network coupling is neuronal cell-subtype specific. Employing in vivo two-photon (2-p) Calcium (Ca) imaging and 2-p targeted whole-cell recordings, we cell-type specifically investigated the coupling profiles of genetically defined neuron populations in superficial layers (L) of mouse primary visual cortex (V1). Our data reveal novel subtlety of neuron-network coupling in inhibitory interneurons (INs). Parvalbumin (PV)- and Vasoactive intestinal peptide (VIP)-expressing INs exhibit skewed distributions towards strong network-coupling; in Somatostatin (SST)-expressing INs, however, two physiological subpopulations are identified with distinct neuron-network coupling profiles, providing direct evidence for subtype specificity. Our results thus add novel functional granularity to neuronal cell-typing, and provided insights critical to simplifying/understanding neural dynamics.Synchronised neuronal activity is essential for cortical function, yet mechanistic insights into this process remain limited. Here, authors use a combination of in vivo imaging and targeted whole-cell recordings to demonstrate that Somatostatin neurons, in the superficial layers of the mouse primary visual cortex, exhibit functional heterogeneity and can be classified into two distinct subtypes characterized as either having type I uncorrelated, or type II highly correlated with network activity.
42543472
Synchronization of cortical activity and its putative role in information processing and learning.
The first measurements of the electrical activity of the brain, performed more than 60 years ago with recordings from the scalp, revealed prominent oscillatory activity (19), and until more recently the analysis of oscillatory patterns in the electroencephalogram and in field potentials recorded with intracerebral macroelectrodes has remained a major research tool of neuro­ physiology. Analyzing the time structure of these wave patterns has estab­ lished close correlations between the frequency sp�ctrum of the oscillations and changes in the central state of the brain. Different stages of sleep and arousal and also abnormal states such as coma and anesthesia could be identified by relying exclusively on the spectral composition of frequencies in the EEG (for review see 14). However, once it became possible to record the activity of individual nerve cells, interest in field potentials and temporal structures declined. The discovery that the firing rate of neurons in peripheral structures of the nervous system reflected the intensity of sensory stimuli and the speed and force of muscle contractions introduced the notion of rate coding. The evidence of a close relation between the position of a neuron in the brain and its functional properties led to the concept of place coding. The
2048816
Correlated neuronal activity and the flow of neural information
For years we have known that cortical neurons collectively have synchronous or oscillatory patterns of activity, the frequencies and temporal dynamics of which are associated with distinct behavioural states. Although the function of these oscillations has remained obscure, recent experimental and theoretical results indicate that correlated fluctuations might be important for cortical processes, such as attention, that control the flow of information in the brain.
205507595
The origin of extracellular fields and currents — EEG, ECoG, LFP and spikes
Neuronal activity in the brain gives rise to transmembrane currents that can be measured in the extracellular medium. Although the major contributor of the extracellular signal is the synaptic transmembrane current, other sources — including Na+ and Ca2+ spikes, ionic fluxes through voltage- and ligand-gated channels, and intrinsic membrane oscillations — can substantially shape the extracellular field. High-density recordings of field activity in animals and subdural grid recordings in humans, combined with recently developed data processing tools and computational modelling, can provide insight into the cooperative behaviour of neurons, their average synaptic input and their spiking output, and can increase our understanding of how these processes contribute to the extracellular signal.
779103
Attention modulates synchronized neuronal firing in primate somatosensory cortex
A potentially powerful information processing strategy in the brain is to take advantage of the temporal structure of neuronal spike trains. An increase in synchrony within the neural representation of an object or location increases the efficacy of that neural representation at the next synaptic stage in the brain; thus, increasing synchrony is a candidate for the neural correlate of attentional selection. We investigated the synchronous firing of pairs of neurons in the secondary somatosensory cortex (SII) of three monkeys trained to switch attention between a visual task and a tactile discrimination task. We found that most neuron pairs in SII cortex fired synchronously and, furthermore, that the degree of synchrony was affected by the monkey's attentional state. In the monkey performing the most difficult task, 35% of neuron pairs that fired synchronously changed their degree of synchrony when the monkey switched attention between the tactile and visual tasks. Synchrony increased in 80% and decreased in 20% of neuron pairs affected by attention.
982790
Internal brain state regulates membrane potential synchrony in barrel cortex of behaving mice
Internal brain states form key determinants for sensory perception, sensorimotor coordination and learning. A prominent reflection of different brain states in the mammalian central nervous system is the presence of distinct patterns of cortical synchrony, as revealed by extracellular recordings of the electroencephalogram, local field potential and action potentials. Such temporal correlations of cortical activity are thought to be fundamental mechanisms of neuronal computation. However, it is unknown how cortical synchrony is reflected in the intracellular membrane potential (Vm) dynamics of behaving animals. Here we show, using dual whole-cell recordings from layer 2/3 primary somatosensory barrel cortex in behaving mice, that the Vm of nearby neurons is highly correlated during quiet wakefulness. However, when the mouse is whisking, an internally generated state change reduces the Vm correlation, resulting in a desynchronized local field potential and electroencephalogram. Action potential activity was sparse during both quiet wakefulness and active whisking. Single action potentials were driven by a large, brief and specific excitatory input that was not present in the Vm of neighbouring cells. Action potential initiation occurs with a higher signal-to-noise ratio during active whisking than during quiet periods. Therefore, we show that an internal brain state dynamically regulates cortical membrane potential synchrony during behaviour and defines different modes of cortical processing.
4409655
Sensory stimulation shifts visual cortex from synchronous to asynchronous states
In the mammalian cerebral cortex, neural responses are highly variable during spontaneous activity and sensory stimulation. To explain this variability, the cortex of alert animals has been proposed to be in an asynchronous high-conductance state in which irregular spiking arises from the convergence of large numbers of uncorrelated excitatory and inhibitory inputs onto individual neurons. Signatures of this state are that a neuron’s membrane potential (Vm) hovers just below spike threshold, and its aggregate synaptic input is nearly Gaussian, arising from many uncorrelated inputs. Alternatively, irregular spiking could arise from infrequent correlated input events that elicit large fluctuations in Vm (refs 5, 6). To distinguish between these hypotheses, we developed a technique to perform whole-cell Vm measurements from the cortex of behaving monkeys, focusing on primary visual cortex (V1) of monkeys performing a visual fixation task. Here we show that, contrary to the predictions of an asynchronous state, mean Vm during fixation was far from threshold (14 mV) and spiking was triggered by occasional large spontaneous fluctuations. Distributions of Vm values were skewed beyond that expected for a range of Gaussian input, but were consistent with synaptic input arising from infrequent correlated events. Furthermore, spontaneous fluctuations in Vm were correlated with the surrounding network activity, as reflected in simultaneously recorded nearby local field potential. Visual stimulation, however, led to responses more consistent with an asynchronous state: mean Vm approached threshold, fluctuations became more Gaussian, and correlations between single neurons and the surrounding network were disrupted. These observations show that sensory drive can shift a common cortical circuitry from a synchronous to an asynchronous state.
4454069
Diverse coupling of neurons to populations in sensory cortex
A large population of neurons can, in principle, produce an astronomical number of distinct firing patterns. In cortex, however, these patterns lie in a space of lower dimension, as if individual neurons were “obedient members of a huge orchestra”. Here we use recordings from the visual cortex of mouse (Mus musculus) and monkey (Macaca mulatta) to investigate the relationship between individual neurons and the population, and to establish the underlying circuit mechanisms. We show that neighbouring neurons can differ in their coupling to the overall firing of the population, ranging from strongly coupled ‘choristers’ to weakly coupled ‘soloists’. Population coupling is largely independent of sensory preferences, and it is a fixed cellular attribute, invariant to stimulus conditions. Neurons with high population coupling are more strongly affected by non-sensory behavioural variables such as motor intention. Population coupling reflects a causal relationship, predicting the response of a neuron to optogenetically driven increases in local activity. Moreover, population coupling indicates synaptic connectivity; the population coupling of a neuron, measured in vivo, predicted subsequent in vitro estimates of the number of synapses received from its neighbours. Finally, population coupling provides a compact summary of population activity; knowledge of the population couplings of n neurons predicts a substantial portion of their n2 pairwise correlations. Population coupling therefore represents a novel, simple measure that characterizes the relationship of each neuron to a larger population, explaining seemingly complex network firing patterns in terms of basic circuit variables.
748328
Petilla terminology: nomenclature of features of GABAergic interneurons of the cerebral cortex
Neuroscience produces a vast amount of data from an enormous diversity of neurons. A neuronal classification system is essential to organize such data and the knowledge that is derived from them. Classification depends on the unequivocal identification of the features that distinguish one type of neuron from another. The problems inherent in this are particularly acute when studying cortical interneurons. To tackle this, we convened a representative group of researchers to agree on a set of terms to describe the anatomical, physiological and molecular features of GABAergic interneurons of the cerebral cortex. The resulting terminology might provide a stepping stone towards a future classification of these complex and heterogeneous cells. Consistent adoption will be important for the success of such an initiative, and we also encourage the active involvement of the broader scientific community in the dynamic evolution of this project.
4396795
Interneuron cell types are fit to function
Understanding brain circuits begins with an appreciation of their component parts — the cells. Although GABAergic interneurons are a minority population within the brain, they are crucial for the control of inhibition. Determining the diversity of these interneurons has been a central goal of neurobiologists, but this amazing cell type has so far defied a generalized classification system. Interneuron complexity within the telencephalon could be simplified by viewing them as elaborations of a much more finite group of developmentally specified cardinal classes that become further specialized as they mature. Our perspective emphasizes that the ultimate goal is to dispense with classification criteria and directly define interneuron types by function.
1546069
Functional specificity of local synaptic connections in neocortical networks
Neuronal connectivity is fundamental to information processing in the brain. Therefore, understanding the mechanisms of sensory processing requires uncovering how connection patterns between neurons relate to their function. On a coarse scale, long-range projections can preferentially link cortical regions with similar responses to sensory stimuli. But on the local scale, where dendrites and axons overlap substantially, the functional specificity of connections remains unknown. Here we determine synaptic connectivity between nearby layer 2/3 pyramidal neurons in vitro, the response properties of which were first characterized in mouse visual cortex in vivo. We found that connection probability was related to the similarity of visually driven neuronal activity. Neurons with the same preference for oriented stimuli connected at twice the rate of neurons with orthogonal orientation preferences. Neurons responding similarly to naturalistic stimuli formed connections at much higher rates than those with uncorrelated responses. Bidirectional synaptic connections were found more frequently between neuronal pairs with strongly correlated visual responses. Our results reveal the degree of functional specificity of local synaptic connections in the visual cortex, and point to the existence of fine-scale subnetworks dedicated to processing related sensory information.
14063660
Dense Inhibitory Connectivity in Neocortex
The connectivity diagram of neocortical circuits is still unknown, and there are conflicting data as to whether cortical neurons are wired specifically or not. To investigate the basic structure of cortical microcircuits, we use a two-photon photostimulation technique that enables the systematic mapping of synaptic connections with single-cell resolution. We map the inhibitory connectivity between upper layers somatostatin-positive GABAergic interneurons and pyramidal cells in mouse frontal cortex. Most, and sometimes all, inhibitory neurons are locally connected to every sampled pyramidal cell. This dense inhibitory connectivity is found at both young and mature developmental ages. Inhibitory innervation of neighboring pyramidal cells is similar, regardless of whether they are connected among themselves or not. We conclude that local inhibitory connectivity is promiscuous, does not form subnetworks, and can approach the theoretical limit of a completely connected synaptic matrix.
152194651
Behavioural report of single neuron stimulation in somatosensory cortex
In welcher Beziehung steht die neuronale Aktivität im Großhirn zur Wahrnehmung von Sinnesreizen? Ist für Wahrnehmungsprozesse das Zusammenwirken zahlreicher aktiver, d.h. Aktionspotenziale feuernder Nervenzellen erforderlich oder haben kleine Neuronengruppen oder sogar einzelne Zellen einen Einfluss auf den Wahrnehmungsprozess? Welche Nervenzelltypen sind hier von besonderer Bedeutung? Dies sind zentrale Fragestellungen der neurowissenschaftlichen Forschung. Schon früh konnte gezeigt werden, dass die Mikrostimulationen weniger Neurone die Perzeption von Sinnesreizen beeinflusst. In einer Publikation aus dem Jahre 2004 zeigten Brecht und Mitarbeiter am motorischen Kortex, dass durch die intrazelluläre Stimulation weniger Aktionspotenziale in einzelnen Pyramidenzellen der Lamina 5 und 6 Schnurrhaarbewegungen ausgelöst werden können. Unbekannt war jedoch bisher, wie sich die Aktivität einzelner Nervenzellen in sensorischen Kortizes auf die Wahrnehmung von Sinnesreizen auswirkt. Arthur R. Houweling und Michael Brecht haben zu dieser Fragestellung jetzt einen wichtigen Beitrag geleistet. Die Forscher aus des Berliner Bernstein Center for Computational Neuroscience führten dazu Experimente am somatosensorischen System, genauer dem Schnurrhaar-‘Barrel’-KortexSystem der Ratte durch. Sie untersuchten den Einfluss, den die Stimulation einzelner Nervenzellen auf ein zuvor gelerntes Verhalten hatte. Dazu wurde folgende Aufgabe trainiert: Kleinere Neuronengruppen wurden in vivo mit einer extrazellulären Mikroelektrode gereizt. Antworteten die Ratten mit einem Zungenlecken, erfolgte eine Belohnung durch Wassergabe. Die Mikrostimulation entspricht dabei der Wahrnehmung eines taktilen Reizes über die Schnurrhaare. Die so trainierten Tiere wurden nun auf ihre Fähigkeit untersucht, die Aktivität eines einzelnen Neurons wahrzunehmen. Um einzelne Neurone zu stimulieren, benutzten die Autoren die so genannte ‘juxtazelluläre’ Stimulation, bei der eine Glaspipette in unmittelbaren Kontakt mit dem Soma des Neurons gebracht wird, sodass durch entsprechend starke Strominjektion (200 ms Dauer) im Mittel etwa 15 Aktionspotenziale in dem untersuchten Neuron ausgelöst werden können; zur gleichzeitigen Mikrostimulation war an der Glaspipette eine Wolfram-Mikroelektrode angebracht um größere Neuronenpopulationen zu stimulieren (Abbildung 1, 2). Es wurden nur Neurone tieferer kortikaler Schichten (der Laminae 4, 5A und 5B) gereizt, wobei sowohl exzitatorische als auch inhibitorische Neurone untersucht wurden. Inhibitorische Neurone unterschieden sich dabei von exzitatorischen Neuronen durch die Aktionspotenzialform und die maximale Feuerfrequenz (wobei inhibitorische Interneurone meist kürzere Aktionspotenziale haben und mit höherer Frequenz feuern können). In einer Versuchsreihe wurde nun entweder mit juxtazellulärer Stimulation, Mikrostimulation oder in Blindkontrollen (ohne oder mit unterschwelliger Strominjektion, sog. ‘catch trial’) gereizt, deren Abfolge zufällig variierte. Das Auftreten einer Verhaltensantwort (dem Lecken) nach Stimulation der Ratte wurde dabei gemessen. Ein derartiges Experiment ist in Abbildung 2 gezeigt. Es zeigte sich, dass nicht nur durch Mikrostimulation bei der Ratte eine Verhaltensantwort ausgelöst werden konnte (in 75% der Fälle), sondern dass auch bei juxtazellulärer Einzelzellstimulation die Tiere häufiger antworteten als bei den Blindversuchen. Dieser Effekt war innerhalb eines Experiments zwar klein, erwies sich jedoch bei Berücksichtigung aller Experimente als statistisch signifikant. In Blindversuchen ohne Stimulation zeigten die Ratten in 18% der Fälle ein Lecken; bei juxtazellulärer Stimulation wurde diese Verhaltensantwort jedoch in 22% beobachtet. Dies bedeutet, dass zumindest in einigen Fällen die Stimulation von einzelnen Neuronen das Verhalten des Tieres beeinflussen kann. Bei den Experimenten von Houweling und Brecht zeigte sich, dass die Einzellzellstimulation von putativ exzitatorischen und inhibitorischen Interneuronen unterschiedliche Folgen für das antrainierte Verhalten hat. Bemerkenswert war, dass zumindest in einigen Fällen die Stimulation von putativen Interneuronen zu besonders häufigen Verhaltensantworten führte. Dies bedeutet, dass die Stimulation von inhibitorischen und exzitatorischen Neuronen unterschiedliche Folgen für die Wahrnehmung von Sinnesreizen oder das Verhalten hat. Auch scheint die Unterdrückung von Aktionspotenzialen bei sensorischen Antworten eine Rolle zu spielen. Es wäre interessant zu untersuchen, inwieweit dieser Effekt auf einem spezifischen Interneurontyp beruht und welche Bedingungen für eine derart starke Antwort erfüllt sein müssen. In der gleichen Ausgabe von Nature veröffentlichten auch Huber et al. eine Abb. 1: Versuchsaufbau und Stimulations-
5730324
Transgenic Mice for Intersectional Targeting of Neural Sensors and Effectors with High Specificity and Performance
UNLABELLED
An increasingly powerful approach for studying brain circuits relies on targeting genetically encoded sensors and effectors to specific cell types. However, current approaches for this are still limited in functionality and specificity. Here we utilize several intersectional strategies to generate multiple transgenic mouse lines expressing high levels of novel genetic tools with high specificity. We developed driver and double reporter mouse lines and viral vectors using the Cre/Flp and Cre/Dre double recombinase systems and established a new, retargetable genomic locus, TIGRE, which allowed the generation of a large set of Cre/tTA-dependent reporter lines expressing fluorescent proteins, genetically encoded calcium, voltage, or glutamate indicators, and optogenetic effectors, all at substantially higher levels than before. High functionality was shown in example mouse lines for GCaMP6, YCX2.60, VSFP Butterfly 1.2, and Jaws. These novel transgenic lines greatly expand the ability to monitor and manipulate neuronal activities with increased specificity.


VIDEO ABSTRACT
16471589
Visual Receptive Field Structure of Cortical Inhibitory Neurons Revealed by Two-Photon Imaging Guided Recording
Synaptic inhibition plays an important role in shaping receptive field (RF) properties in the visual cortex. However, the underlying mechanisms remain not well understood, partly because of difficulties in systematically studying functional properties of cortical inhibitory neurons in vivo. Here, we established two-photon imaging guided cell-attached recordings from genetically labeled inhibitory neurons and nearby “shadowed” excitatory neurons in the primary visual cortex of adult mice. Our results revealed that in layer 2/3, the majority of excitatory neurons exhibited both On and Off spike subfields, with their spatial arrangement varying from being completely segregated to overlapped. In contrast, most layer 4 excitatory neurons exhibited only one discernable subfield. Interestingly, no RF structure with significantly segregated On and Off subfields was observed for layer 2/3 inhibitory neurons of either the fast-spike or regular-spike type. They predominantly possessed overlapped On and Off subfields with a significantly larger size than the excitatory neurons and exhibited much weaker orientation tuning. These results from the mouse visual cortex suggest that different from the push-pull model proposed for simple cells, layer 2/3 simple-type neurons with segregated spike On and Off subfields likely receive spatially overlapped inhibitory On and Off inputs. We propose that the phase-insensitive inhibition can enhance the spatial distinctiveness of On and Off subfields through a gain control mechanism.
8066170
Broadly Tuned Response Properties of Diverse Inhibitory Neuron Subtypes in Mouse Visual Cortex
Different subtypes of GABAergic neurons in sensory cortex exhibit diverse morphology, histochemical markers, and patterns of connectivity. These subtypes likely play distinct roles in cortical function, but their in vivo response properties remain unclear. We used in vivo calcium imaging, combined with immunohistochemical and genetic labels, to record visual responses in excitatory neurons and up to three distinct subtypes of GABAergic neurons (immunoreactive for parvalbumin, somatostatin, or vasoactive intestinal peptide) in layer 2/3 of mouse visual cortex. Excitatory neurons had sharp response selectivity for stimulus orientation and spatial frequency, while all GABAergic subtypes had broader selectivity. Further, bias in the responses of GABAergic neurons toward particular orientations or spatial frequencies tended to reflect net biases of the surrounding neurons. These results suggest that the sensory responses of layer 2/3 GABAergic neurons reflect the pooled activity of the surrounding population--a principle that may generalize across species and sensory modalities.
13907138
Somatostatin-expressing neurons in cortical networks
Somatostatin-expressing GABAergic neurons constitute a major class of inhibitory neurons in the mammalian cortex and are characterized by dense wiring into the local network and high basal firing activity that persists in the absence of synaptic input. This firing provides both GABA type A receptor (GABAAR)- and GABABR-mediated inhibition that operates at fast and slow timescales. The activity of somatostatin-expressing neurons is regulated by brain state, during learning and in rewarded behaviour. Here, we review recent advances in our understanding of how this class of cells can control network activity, with specific reference to how this is constrained by their anatomical and electrophysiological properties.
195683623
Differential connectivity and response dynamics of excitatory and inhibitory neurons in visual cortex.
Neuronal responses during sensory processing are influenced by both the organization of intracortical connections and the statistical features of sensory stimuli. How these intrinsic and extrinsic factors govern the activity of excitatory and inhibitory populations is unclear. Using two-photon calcium imaging in vivo and intracellular recordings in vitro, we investigated the dependencies between synaptic connectivity, feature selectivity and network activity in pyramidal cells and fast-spiking parvalbumin-expressing (PV) interneurons in mouse visual cortex. In pyramidal cell populations, patterns of neuronal correlations were largely stimulus-dependent, indicating that their responses were not strongly dominated by functionally biased recurrent connectivity. By contrast, visual stimulation only weakly modified co-activation patterns of fast-spiking PV cells, consistent with the observation that these broadly tuned interneurons received very dense and strong synaptic input from nearby pyramidal cells with diverse feature selectivities. Therefore, feedforward and recurrent network influences determine the activity of excitatory and inhibitory ensembles in fundamentally different ways.
4423189
Driving fast-spiking cells induces gamma rhythm and controls sensory responses
Cortical gamma oscillations (20-80 Hz) predict increases in focused attention, and failure in gamma regulation is a hallmark of neurological and psychiatric disease. Current theory predicts that gamma oscillations are generated by synchronous activity of fast-spiking inhibitory interneurons, with the resulting rhythmic inhibition producing neural ensemble synchrony by generating a narrow window for effective excitation. We causally tested these hypotheses in barrel cortex in vivo by targeting optogenetic manipulation selectively to fast-spiking interneurons. Here we show that light-driven activation of fast-spiking interneurons at varied frequencies (8-200 Hz) selectively amplifies gamma oscillations. In contrast, pyramidal neuron activation amplifies only lower frequency oscillations, a cell-type-specific double dissociation. We found that the timing of a sensory input relative to a gamma cycle determined the amplitude and precision of evoked responses. Our data directly support the fast-spiking-gamma hypothesis and provide the first causal evidence that distinct network activity states can be induced in vivo by cell-type-specific activation.
6192188
Not all that glitters is gold: off-target recombination in the somatostatin–IRES-Cre mouse line labels a subset of fast-spiking interneurons
Neurons in the mammalian brain are a highly diverse population with a complex assortment of electrophysiological, morphological and molecular properties, which has hindered efforts to classify them into genetically and functionally meaningful subtypes and to understand their various roles in the normal or pathological brain. Nowhere is this issue more acutely felt than in the study of inhibitory cortical interneurons, whose classification is still a source of contention and confusion (Markram et al., 2004; Petilla Interneuron Nomenclature et al., 2008; Defelipe et al., 2013). A major boon to investigators has been the development of mouse lines in which genetically defined subsets of interneurons express fluorescent proteins, allowing their identification and targeting during electrophysiological recordings or imaging experiments (Oliva et al., 2000; Meyer et al., 2002; Chattopadhyaya et al., 2004; Ma et al., 2006). More recently, investigators funded by the NIH Neuroscience Blueprint project have developed a toolbox of “driver” lines in which the Cre recombinase gene is inserted immediately downstream to genes that are known markers of specific interneuron subsets (Taniguchi et al., 2011). These Cre lines can be bred with mice carrying floxed genes, to generate cell type-specific knockouts of genes of interest. In addition, by breeding these lines with mice from a parallel toolbox of Cre reporter lines in which a gene coding sequence is inserted after a lox-STOP-lox cassette, or by transfecting them with viral vectors carrying similar constructs, investigators can induce cell-type specific expression of any gene of interest, from inert fluorescent proteins to calcium probes and light-activated ion channels or pumps (Madisen et al., 2010, 2012; Zariwala et al., 2012). While these technologies carry great promise and have already enabled some important findings, the rush to use them also carries considerable risk, if the relevant expression patterns are not fully characterized. A case in point is the somatostatin–IRES-Cre (SOM-Cre) mouse line (Taniguchi et al., 2011), in which Cre expression was targeted to cells containing the neuropeptide somatostatin (SOM). In the cerebral cortex, SOM-containing neurons are a well-studied population of dendritic-targeting inhibitory interneurons (Ma et al., 2006; Silberberg and Markram, 2007; Fanselow et al., 2008; Tan et al., 2008; Ma et al., 2010; Fino and Yuste, 2011). The SOM-Cre line has already been used in several high-profile studies, and in most of these the authors tacitly assumed—but did not validate—that Cre-mediated recombination was restricted to SOM interneurons (Adesnik et al., 2012; Gentet et al., 2012; Lee et al., 2012; Wilson et al., 2012; Chiu et al., 2013; Kvitsiani et al., 2013; Xu et al., 2013). We found, however, that 6–10% of neurons expressing a Cre-dependent reporter in any given cortical layer were fast-spiking/ parvalbumin-expressing (FS/PV) interneurons, a subtype quite distinct from SOM interneurons in electrophysiological etc., morphological and molecular properties (Rudy et al., 2011) [Note that there is another SOM-Cre line reported in the literature (Lovett-Barron et al., 2012), which we did not test].
17527528
Cellular mechanisms of brain-state-dependent gain modulation in visual cortex
Visual cortical neurons fire at higher rates to visual stimuli during locomotion than during immobility, while maintaining orientation selectivity. The mechanisms underlying this change in gain are not understood. We performed whole-cell recordings from layer 2/3 and layer 4 visual cortical excitatory neurons and from parvalbumin-positive and somatostatin-positive inhibitory neurons in mice that were free to rest or run on a spherical treadmill. We found that the membrane potential of all cell types became more depolarized and (with the exception of somatostatin-positive interneurons) less variable during locomotion. Cholinergic input was essential for maintaining the unimodal membrane potential distribution during immobility, whereas noradrenergic input was necessary for the tonic depolarization associated with locomotion. Our results provide a mechanism for how neuromodulation controls the gain and signal-to-noise ratio of visual cortical neurons during changes in the state of vigilance.
599889
A Cortical Circuit for Gain Control by Behavioral State
The brain's response to sensory input is strikingly modulated by behavioral state. Notably, the visual response of mouse primary visual cortex (V1) is enhanced by locomotion, a tractable and accessible example of a time-locked change in cortical state. The neural circuits that transmit behavioral state to sensory cortex to produce this modulation are unknown. In vivo calcium imaging of behaving animals revealed that locomotion activates vasoactive intestinal peptide (VIP)-positive neurons in mouse V1 independent of visual stimulation and largely through nicotinic inputs from basal forebrain. Optogenetic activation of VIP neurons increased V1 visual responses in stationary awake mice, artificially mimicking the effect of locomotion, and photolytic damage of VIP neurons abolished the enhancement of V1 responses by locomotion. These findings establish a cortical circuit for the enhancement of visual response by locomotion and provide a potential common circuit for the modulation of sensory processing by behavioral state.
4713941
Vision and Locomotion Shape the Interactions between Neuron Types in Mouse Visual Cortex
Summary Cortical computation arises from the interaction of multiple neuronal types, including pyramidal (Pyr) cells and interneurons expressing Sst, Vip, or Pvalb. To study the circuit underlying such interactions, we imaged these four types of cells in mouse primary visual cortex (V1). Our recordings in darkness were consistent with a “disinhibitory” model in which locomotion activates Vip cells, thus inhibiting Sst cells and disinhibiting Pyr cells. However, the disinhibitory model failed when visual stimuli were present: locomotion increased Sst cell responses to large stimuli and Vip cell responses to small stimuli. A recurrent network model successfully predicted each cell type’s activity from the measured activity of other types. Capturing the effects of locomotion, however, required allowing it to increase feedforward synaptic weights and modulate recurrent weights. This network model summarizes interneuron interactions and suggests that locomotion may alter cortical computation by changing effective synaptic connectivity.
594875
Unique functional properties of somatostatin-expressing GABAergic neurons in mouse barrel cortex
Neocortical GABAergic neurons have diverse molecular, structural and electrophysiological features, but the functional correlates of this diversity are largely unknown. We found unique membrane potential dynamics of somatostatin-expressing (SOM) neurons in layer 2/3 of the primary somatosensory barrel cortex of awake behaving mice. SOM neurons were spontaneously active during periods of quiet wakefulness. However, SOM neurons hyperpolarized and reduced action potential firing in response to both passive and active whisker sensing, in contrast with all other recorded types of nearby neurons, which were excited by sensory input. Optogenetic inhibition of SOM neurons increased burst firing in nearby excitatory neurons. We hypothesize that the spontaneous activity of SOM neurons during quiet wakefulness provides a tonic inhibition to the distal dendrites of excitatory pyramidal neurons. Conversely, the inhibition of SOM cells during active cortical processing likely enhances distal dendritic excitability, which may be important for top-down computations and sensorimotor integration.
4341820
Long-term in vivo imaging of experience-dependent synaptic plasticity in adult cortex
Do new synapses form in the adult cortex to support experience-dependent plasticity? To address this question, we repeatedly imaged individual pyramidal neurons in the mouse barrel cortex over periods of weeks. We found that, although dendritic structure is stable, some spines appear and disappear. Spine lifetimes vary greatly: stable spines, about 50% of the population, persist for at least a month, whereas the remainder are present for a few days or less. Serial-section electron microscopy of imaged dendritic segments revealed retrospectively that spine sprouting and retraction are associated with synapse formation and elimination. Experience-dependent plasticity of cortical receptive fields was accompanied by increased synapse turnover. Our measurements suggest that sensory experience drives the formation and elimination of synapses and that these changes might underlie adaptive remodelling of neural circuits.
6763838
Dendritic Inhibition in the Hippocampus Supports Fear Learning
Fear, Memory, and Place Contextual fear conditioning (CFC) is widely used as a hippocampal-dependent classical conditioning task to model human episodic memory. Lovett-Barron et al. (p. 857) combined in vivo imaging with pharmacology, pharmacogenetics, and optogenetics and they found that somatostatin-expressing, dendrite-targeting γ-aminobutyric acid–releasing interneurons in hippocampal area CA1 are required for CFC. During CFC, sensory features of the aversive event reach hippocampal output neurons through excitatory cortical afferents and require active inhibitory filtering to ensure that the hippocampus exclusively encodes the conditioned stimulus. Cholinergic activation of somatostatin-positive hippocampal CA1 interneurons promotes fear-context associations. Fear memories guide adaptive behavior in contexts associated with aversive events. The hippocampus forms a neural representation of the context that predicts aversive events. Representations of context incorporate multisensory features of the environment, but must somehow exclude sensory features of the aversive event itself. We investigated this selectivity using cell type–specific imaging and inactivation in hippocampal area CA1 of behaving mice. Aversive stimuli activated CA1 dendrite-targeting interneurons via cholinergic input, leading to inhibition of pyramidal cell distal dendrites receiving aversive sensory excitation from the entorhinal cortex. Inactivating dendrite-targeting interneurons during aversive stimuli increased CA1 pyramidal cell population responses and prevented fear learning. We propose subcortical activation of dendritic inhibition as a mechanism for exclusion of aversive stimuli from hippocampal contextual representations during fear learning.
205242868
Branch-specific dendritic Ca2+ spikes cause persistent synaptic plasticity
The brain has an extraordinary capacity for memory storage, but how it stores new information without disrupting previously acquired memories remains unknown. Here we show that different motor learning tasks induce dendritic Ca2+ spikes on different apical tuft branches of individual layer V pyramidal neurons in the mouse motor cortex. These task-related, branch-specific Ca2+ spikes cause long-lasting potentiation of postsynaptic dendritic spines active at the time of spike generation. When somatostatin-expressing interneurons are inactivated, different motor tasks frequently induce Ca2+ spikes on the same branches. On those branches, spines potentiated during one task are depotentiated when they are active seconds before Ca2+ spikes induced by another task. Concomitantly, increased neuronal activity and performance improvement after learning one task are disrupted when another task is learned. These findings indicate that dendritic-branch-specific generation of Ca2+ spikes is crucial for establishing long-lasting synaptic plasticity, thereby facilitating information storage associated with different learning experiences.
2113877
A cortical disinhibitory circuit for enhancing adult plasticity
The adult brain continues to learn and can recover from injury, but the elements and operation of the neural circuits responsible for this plasticity are not known. In previous work, we have shown that locomotion dramatically enhances neural activity in the visual cortex (V1) of the mouse (Niell and Stryker, 2010), identified the cortical circuit responsible for this enhancement (Fu et al., 2014), and shown that locomotion also dramatically enhances adult plasticity (Kaneko and Stryker, 2014). The circuit that is responsible for enhancing neural activity in the visual cortex contains both vasoactive intestinal peptide (VIP) and somatostatin (SST) neurons (Fu et al., 2014). Here, we ask whether this VIP-SST circuit enhances plasticity directly, independent of locomotion and aerobic activity. Optogenetic activation or genetic blockade of this circuit reveals that it is both necessary and sufficient for rapidly increasing V1 cortical responses following manipulation of visual experience in adult mice. These findings reveal a disinhibitory circuit that regulates adult cortical plasticity. DOI: http://dx.doi.org/10.7554/eLife.05558.001
205418185
Targeted patch-clamp recordings and single-cell electroporation of unlabeled neurons in vivo
Here we describe an approach for making targeted patch-clamp recordings from single neurons in vivo, visualized by two-photon microscopy. A patch electrode is used to perfuse the extracellular space surrounding the neuron of interest with a fluorescent dye, thus enabling the neuron to be visualized as a negative image ('shadow') and identified on the basis of its somatodendritic structure. The same electrode is then placed on the neuron under visual control to allow formation of a gigaseal ('shadowpatching'). We demonstrate the reliability and versatility of shadowpatching by performing whole-cell recordings from visually identified neurons in the neocortex and cerebellum of rat and mouse. We also show that the method can be used for targeted in vivo single-cell electroporation of plasmid DNA into identified cell types, leading to stable transgene expression. This approach facilitates the recording, labeling and genetic manipulation of single neurons in the intact native mammalian brain without the need to pre-label neuronal populations.
9009449
Targeted Whole-Cell Recordings in the Mammalian Brain In Vivo
While electrophysiological recordings from visually identified cell bodies or dendrites are routinely performed in cell culture and acute brain slice preparations, targeted recordings from the mammalian nervous system are currently not possible in vivo. The "blind" approach that is used instead is somewhat random and largely limited to common neuronal cell types. This approach prohibits recordings from, for example, molecularly defined and/or disrupted populations of neurons. Here we describe a method, which we call TPTP (two-photon targeted patching), that uses two-photon imaging to guide in vivo whole-cell recordings to individual, genetically labeled cortical neurons. We apply this technique to obtain recordings from genetically manipulated, parvalbumin-EGFP-positive interneurons in the somatosensory cortex. We find that both spontaneous and sensory-evoked activity patterns involve the synchronized discharge of electrically coupled interneurons. TPTP applied in vivo will therefore provide new insights into the molecular control of neuronal function at the systems level.
120456150
Measures of multivariate skewness and kurtosis with applications
SUMMARY Measures of multivariate skewness and kurtosis are developed by extending certain studies on robustness of the t statistic. These measures are shown to possess desirable properties. The asymptotic distributions of the measures for samples from a multivariate normal population are derived and a test of multivariate normality is proposed. The effect of nonnormality on the size of the one-sample Hotelling's T2 test is studied empirically with the help of these measures, and it is found that Hotelling's T2 test is more sensitive to the measure of skewness than to the measure of kurtosis. measures have proved useful (i) in selecting a member of a family such as from the Karl Pearson family, (ii) in developing a test of normality, and (iii) in investigating the robustness of the standard normal theory procedures. The role of the tests of normality in modern statistics has recently been summarized by Shapiro & Wilk (1965). With these applications in mind for the multivariate situations, we propose measures of multivariate skewness and kurtosis. These measures of skewness and kurtosis are developed naturally by extending certain aspects of some robustness studies for the t statistic which involve I1 and 32. It should be noted that measures of multivariate dispersion have been available for quite some time (Wilks, 1932, 1960; Hotelling, 1951). We deal with the measure of skewness in ? 2 and with the measure of kurtosis in ? 3. In ? 4 we give two important applications of these measures, namely, a test of multivariate normality and a study of the effect of nonnormality on the size of the one-sample Hotelling's T2 test. Both of these problems have attracted attention recently. The first problem has been treated by Wagle (1968) and Day (1969) and the second by Arnold (1964), but our approach differs from theirs.