Chemosignal transduction in the vomeronasal organ of garter snakes: cloning of a gene encoding adenylate cyclase from the vomeronasal organ of garter snakes.

We previously reported that ES20-receptor binding activates phosphoinositide (PI) turnover, resulting in an increase in inositol-1,4,5-trisphosphate, which in turn mobilizes intracellularly stored calcium in the vomeronasal (VN) sensory epithelium of garter snakes. We also found that the activity of adenylate cyclase (AC) in the VN organ is very sensitive to Ca2+ but insensitive to calmodulin regulation. A 250-bp fragment of adenylate cyclase type VI (AC-VI) was obtained from brain cDNA of garter snake by RT-PCR with degenerate primers. The 250-bp fragments were amplified, cloned, and sequenced. Both Northern blot and RNase protection assays revealed that the vomeronasal organ (VNO) and brain contained more abundance of AC type VI than the main olfactory epithelium. A 3.8-kb cDNA was then cloned from the vomeronasal cDNA library of garter snakes and sequenced. The 5' cDNA was obtained by means of 5' RACE PCR and sequenced. We have successfully cloned a 5200-nucleotide cDNA from VNO of garter snakes containing an open reading frame++ encoding 1150 amino acids of AC-VI protein. The vomeronasal AC is termed AC(VN) . AC(VN) shows a high degree of homology with type VI AC of rat, mouse, or human. In situ hybridization with digoxigenin-labeled cRNA demonstrated that AC(VN) mRNA was abundant in the sensory epithelium but not in the nonsensory epithelium of the mushroom body of the vomeronasal organ of garter snakes.