cAMP-dependent protein kinase A mediation of vasopressin gene expression in the hypothalamus of the osmotically challenged rat.
We have tested the hypothesis that 3', 5'-cyclic adenosine monophosphate (cAMP)-dependent protein kinase A (PKA) is involved in the regulation of the vasopressin (VP) gene in the magnocellular neurons of the paraventricular nucleus (PVN) of the osmotically challenged rat. An adenoviral vector expressing a potent peptide inhibitor of PKA, Ad.CMV.PKIalpha, was demonstrated to be highly efficient in vitro. Ad.CMV.PKIalpha was then introduced into the PVN of rats bearing a VP reporter transgene (3-VCAT-3) consisting of the VP structural gene containing an epitope reporter in exon III, flanked by 3 kb of upstream and 3 kb of downstream sequence Robust transgene expression is seen in VP neurons of the PVN, and this increases following 72 h of dehydration. Ad.CMV.PKIalpha significantly blunted 3-VCAT-3 expression in the osmotically stimulated PVN. Our evidence suggests that PKA mediates changes in VP gene expression in response to dehydration through sequences contained within the 3-VCAT-3 transgene.
Wong
Liang-Fong
LF
The University Research Centre for Neuroendocrinology, University of Bristol, Bristol Royal Infirmary, Marlborough Street, Bristol BS2 8HW, England, UK.
Harding
Tom
T
Uney
James
J
Murphy
David
D
D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D030801:Animals, Genetically Modified; D002352:Carrier Proteins / Q000235:genetics / Q000494:pharmacology; D002460:Cell Line; D017868:Cyclic AMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D003681:Dehydration / Q000235:genetics / Q000378:metabolism; D005260:Female; D005786:Gene Expression Regulation / Q000502:physiology*; D017930:Genes, Reporter / Q000235:genetics; D005822:Genetic Vectors / Q000494:pharmacology; D007030:Hypothalamo-Hypophyseal System / Q000201:enzymology* / Q000378:metabolism; D008297:Male; D010286:Paraventricular Hypothalamic Nucleus / Q000201:enzymology* / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D011993:Recombinant Fusion Proteins / Q000494:pharmacology; D019076:Transgenes / Q000235:genetics; D014667:Vasopressins / Q000096:biosynthesis / Q000235:genetics*; D014882:Water-Electrolyte Balance / Q000235:genetics*
D048868:Adaptor Proteins, Signal Transducing; D002352:Carrier Proteins; C491525:Pkia protein, rat; D011993:Recombinant Fusion Proteins; D014667:Vasopressins; D017868:Cyclic AMP-Dependent Protein Kinases