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Analysis of the Proenkephalin Second Messenger‐Inducible Enhancer in Rat Striatal Cultures
Author(s) -
Konradi Christine,
Cole Rebecca L.,
Green Donnella,
Senatus Patrick,
Leveque JeanChristophe,
Pollack Alexia E.,
Grossbard Sarah J.,
Hyman Steven E.
Publication year - 1995
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1046/j.1471-4159.1995.65031007.x
Subject(s) - proenkephalin , enhancer , second messenger system , messenger rna , biology , neuroscience , microbiology and biotechnology , chemistry , endocrinology , medicine , genetics , gene expression , gene , enkephalin , signal transduction , receptor , opioid
We have previously shown that in cell extracts from rat striatum, cyclic AMP response element (CRE) binding protein (CREB), rather than AP‐1 proteins, preferentially interacts with the CRE‐2 element of the proenkephalin second messenger‐inducible enhancer, even under conditions in which AP‐1 proteins are highly induced. Here we use primary striatal cultures to permit a more detailed analysis of CRE‐2 function and protein binding in relevant neural cell types. By transfection we find that in primary striatal cultures, as in transformed cell lines, the CRE‐1 and CRE‐2 elements are required for significant induction by cyclic AMP. We report that cyclic AMP induction of the proenkephalin gene in striatal cultures is protein synthesis independent, excluding a role for newly synthesized proteins like c‐Fos. We also show that cyclic AMP induces CREB phosphorylation and that phosphorylated CREB interacts strongly with CRE‐2 and weakly with CRE‐1. The predominant protein bound to CRE‐1 is not CREB, however, and remains to be identified. Despite some prior predictions, we do not find a role for c‐Fos in cyclic AMP regulation of proenkephalin gene expression in neurons.