Abstract
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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adaptor Proteins, Signal Transducing
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Animals
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Animals, Genetically Modified
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Carrier Proteins / genetics
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Carrier Proteins / pharmacology
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Cell Line
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Cyclic AMP-Dependent Protein Kinases / antagonists & inhibitors
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Cyclic AMP-Dependent Protein Kinases / metabolism*
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Dehydration / genetics
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Dehydration / metabolism
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Female
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Gene Expression Regulation / physiology*
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Genes, Reporter / genetics
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Genetic Vectors / pharmacology
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Hypothalamo-Hypophyseal System / enzymology*
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Hypothalamo-Hypophyseal System / metabolism
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Male
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Paraventricular Hypothalamic Nucleus / enzymology*
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Paraventricular Hypothalamic Nucleus / metabolism
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Rats
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Rats, Sprague-Dawley
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Recombinant Fusion Proteins / pharmacology
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Transgenes / genetics
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Vasopressins / biosynthesis
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Vasopressins / genetics*
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Water-Electrolyte Balance / genetics*
Substances
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Adaptor Proteins, Signal Transducing
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Carrier Proteins
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Pkia protein, rat
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Recombinant Fusion Proteins
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Vasopressins
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Cyclic AMP-Dependent Protein Kinases