The metabotropic glutamate receptor subtype 5 (mGluR5) is densely expressed in striatal projection neurons. As a G protein-coupled receptor, mGluR5 may initiate an intracellular cascade that conveys extracellular signals to gene expression. This study investigated the possible role of mGluR5 in the inducible phosphorylation of a nuclear transcription factor, cAMP response element-binding protein (CREB), in primary cultures of striatal neurons from rat E19 embryos or neonatal day-1 pups. We found that selective activation of mGluR5 with a selective agonist (RS)-2-chloro-5-hydroxyphenylglycine (CHPG) induced a rapid and transient increase in phosphorylated CREB immunoreactivity in striatal neurons as analyzed by both immunocytochemistry and western blot. The increase in CREB phosphorylation was concentration-dependent, and seen in neurochemically identified GABAergic neurons. Pre-treatment with the mGluR5 antagonist 2-methyl-6-(phenylethynyl)pyridine hydrochloride (MPEP) blocked the CHPG phosphorylation of CREB. In contrast, the mGluR1 antagonist, 7-(hydroxyimino)cyclopropa[b]chromen-1a-carboxylate ethyl ester (CPCCOet) did not alter CHPG-stimulated CREB phosphorylation. The mGluR5 antisense oligonucleotides, but not their controls, selectively reduced basal mGluR5 levels as well as CREB phosphorylation in response to CHPG addition. Lastly, using an immediate early gene c-fos as a reporter of inducible gene expression downstream to phosphorylated CREB, we found that CHPG induced a rapid and transient increase in c-fos mRNA levels in cultured neurons as revealed by quantitative in situ hybridization. The increase in c-fos was kinetically correlated well with the CREB phosphorylation and blocked by MPEP and the CREB antisense oligonucleotides. These results demonstrate a positive linkage from surface mGluR5 to CREB phosphorylation, which is able to facilitate immediate gene expression in striatal neurons.