Expression of metabotropic GABA(B) receptors is essential for slow inhibitory synaptic transmission in the CNS, and disruption of GABA(B) receptor-mediated responses has been associated with several disorders, including neuropathic pain and epilepsy. The location of GABA(B) receptors in neurons determines their specific role in synaptic transmission, and it is believed that sorting of subunit isoforms, GABA(B)R1a and GABA(B)R1b, to presynaptic or postsynaptic membranes helps to determine this role. GABA(B)R1a and GABA(B)R1b are thought to arise by alternative splicing of heteronuclear RNA. We now demonstrate that alternative promoters, rather than alternative splicing, produce GABA(B)R1a and GABA(B)R1b isoforms. Our data further show that subunit gene expression in hippocampal neurons is mediated by the cAMP response element-binding protein (CREB) by binding to unique cAMP response elements in the alternative promoter regions. Double-stranded oligonucleotide decoys selectively alter levels of endogenous GABA(B)R1a and GABA(B)R1b in primary hippocampal neurons, and CREB knock-out mice show changes in levels of GABA(B)R1a and GABA(B)R1b transcripts, consistent with decoy competition experiments. These results demonstrate a critical role of CREB in transcriptional mechanisms that control GABA(B)R1 subunit levels in vivo. In addition, the CREB-related factor activating transcription factor-4 (ATF4) has been shown to interact directly with GABA(B)R1 in neurons, and we show that ATF4 differentially regulates GABA(B)R1a and GABA(B)R1b promoter activity. These results, together with our finding that the depolarization-sensitive upstream stimulatory factor (USF) binds to a composite CREB/ATF4/USF regulatory element only in the absence of CREB binding, indicate that selective control of alternative GABA(B)R1 promoters by CREB, ATF4, and USF may dynamically regulate expression of their gene products in the nervous system.