Differential regulation of AP-1 DNA binding activity in rat hypothalamus and pituitary by estrogen.
Ap-1 proteins such as Fos and Jun are nuclear transcription factors that have been postulated to function as third messengers in signal transduction pathways to regulate target gene expression. Using electrophoretic mobility shift assays (EMSA), we have studied estrogen (E) effects on regulation of AP-1 DNA binding activity in the rat hypothalamus and pituitary. AP-1 binding is defined herein as the specific association with a consensus AP-1 site during EMSA. Specific AP-1 binding activity was observed in nuclear extracts from the hypothalamus and pituitary of ovariectomized (OVX) female and castrated (CAS) male rats. Treatment with E increased the levels of AP-1 binding activity in the pituitary and uterus, whereas E decreased the levels of AP-1 binding in the hypothalamus, of OVX females. These effects were observed within 60 min and maintained for at least 72 h after a single dose of estrogen. Estrogen-induced changes in AP-1 binding were much more prominent in OVX females than in CAS males. Treatment with progesterone in OVX females had no significant effects on AP-1 binding activity in either pituitary or hypothalamus. Analysis of AP-1 binding activity in both hypothalamus and pituitary by supershift, immunodepletion and shift-Western blot indicated that part of the AP-1 binding was due to the presence of Fos and Jun proteins. However, Western blot analysis shows that the levels of Fos and Jun proteins in the hypothalamic nuclear extracts were not altered by E treatment. We conclude that E produced tissue and sex-differentiated alterations in AP-1 DNA binding activity in the hypothalamus and pituitary of female rats, which may be related to differential estrogenic actions on gene regulation.