Alterations of AP-1 and CREB protein DNA binding in rat supraoptic and paraventricular nuclei by acute and repeated
1Department of Anatomy, Physiology and Genetics, F. E. Hébert School of Medicine, The Uniformed Services University of the Health Sciences, Bethesda, MD 20814, USA. jmccabe@usuhs.mil
Abstract:
Electrophoretic mobility shift assays were used to analyze Fos and CREB protein-DNA-interactions in the rat hypothalamic paraventricular (PVN) and supraoptic (SON) nuclei. After intraperitoneal administration of normal saline, PVN (but not SON) extracts exhibited a significant 183% increase in binding to the activational protein-1 (AP-1) canonical DNA binding sequence. Hypertonic saline treatment resulted in a approximately 2.5-fold increase in binding by tissue samples from both regions. AP-1 binding by SON extracts after two hypertonic saline injections caused a 307% increase in binding that was significantly greater than binding by PVN extracts (207%). Fos binding was equal in the SON after one and two hypertonic saline injections, but the PVN exhibited less of an increase after two injections. Binding to the canonical cyclic adenosine monophosphate regulatory element (CRE), and phosphorylated CREB (pCREB) supershift binding, indicated pCREB is constitutively expressed. Any experimental treatment (handling and an injection) caused an elevation in binding in the PVN. AP-1 protein complex DNA binding was increased after osmotic stimulation, and SON and PVN exhibit differences in AP-1 DNA binding kinetics, after repeated hypertonic saline stress. Changes in PVN tissue samples were subtle, and may reflect the fact that magnocellular and parvocellular neurons mediate, respectively, fluid homeostasis and stress responses.
