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Division of Gastrointestinal Hormones, Intrahypothalamic (IHT) administration of neuropeptide Y (NPY) induces
a robust feeding response in rats. We have shown previously that
NPY-induced feeding is mediated by a pertussis-toxin-sensitive G
protein in rats. NPY receptors are coupled to cAMP and
Ca2+. Because these second messengers are known to activate
cAMP response element binding proteins, (CREB), cAMP response element
modulators, or activating transcription factor 1, we investigated the
involvement of these transcription factors in NPY-induced feeding in
rats. Compared with control injections of cerebrospinal fluid (1 µl), IHT administration of NPY increased cAMP response element (CRE) binding
to rat hypothalamic nuclear extracts in a time-dependent manner, as
detected by an electrophoretic mobility shift assay. In contrast, IHT
administration of the anorectic neuropeptide, pituitary adenylate
cyclase activating polypeptide, strongly inhibited the CRE binding.
Food deprivation for 48 hr also increased CRE binding, whereas 8 hr of
refeeding normalized CRE activity. Preincubation of the hypothalamic
nuclear extracts of NPY-treated and unfed rats with antibody specific
to CREB blocked CRE binding, whereas preincubation with phosphoCREB
antibody retarded the migration of CRE-protein complex, indicating that
phosphoCREB is involved in this process. Consistently,
immunohistochemical studies with food-deprived rats showed an intense
phosphoCREB signal in the paraventricular nuclei and ventromedial
hypothalamus in comparison to rats fed ad libitum.
Hypothalamic calcium/calmodulin-dependent protein kinase II activity
was also increased by IHT-NPY. These results suggest that
calcium/calmodulin-dependent protein kinase II induced phosphorylation
of CREB may be involved in regulating feeding behavior induced by NPY.
Copyright © by The American Society for Pharmacology and Experimental Therapeutics
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