Vomeronasal sensory neurons fromSternotherus odoratus(stinkpot/musk turtle) respond to chemosignalsviathe phospholipase C system

Abstract

SUMMARYThe mammalian signal transduction apparatus utilized by vomeronasal sensory neurons (VSNs) in the vomeronasal organ (VNO) has been richly explored, while that of reptiles, and in particular, the stinkpot or musk turtle Sternotherus odoratus, is less understood. Given that the turtle's well-known reproductive and mating behaviors are governed by chemical communication, 247 patch-clamp recordings were made from male and female S. odoratus VSNs to study the chemosignal-activated properties as well as the second-messenger system underlying the receptor potential. Of the total neurons tested, 88 (35%) were responsive to at least one of five complex natural chemicals, some of which demonstrated a degree of sexual dimorphism in response selectivity. Most notably, male VSNs responded to male urine with solely outward currents. Ruthenium Red, an IP3 receptor(IP3R) antagonist, failed to block chemosignal-activated currents,while the phospholipase C (PLC) inhibitor, U73122, abolished the chemosignal-activated current within 2 min, implicating the PLC system in the generation of a receptor potential in the VNO of musk turtles. Dialysis of several second messengers or their analogues failed to elicit currents in the whole-cell patch-clamp configuration, negating a direct gating of the transduction channel by cyclic adenosine monophosphate (cAMP), inositol 1,4,5-trisphosphate (IP3), arachidonic acid (AA), or diacylglycerol(DAG). Reversal potential analysis of chemosignal-evoked currents demonstrated that inward currents reversed at –5.7±7.8 mV (mean ±s.e.m.; N=10), while outward currents reversed at–28.2±2.4 mV (N=30). Measurements of conductance changes associated with outward currents indicated that the outward current represents a reduction of a steady state inward current by the closure of an ion channel when the VSN is exposed to a chemical stimulus such as male urine. Chemosignal-activated currents were significantly reduced when a peptide mimicking a domain on canonical transient receptor potential 2 (TRPC2), to which type 3 IP3 receptor (IP3R3) binds, was included in the recording pipette. Collectively these data suggest that there are multiple transduction cascades operational in the VSNs of S. odoratus, one of which may be mediated by a non-selective cation conductance that is not gated by IP3 but may be modulated by the interaction of its receptor with the TRPC2 channel.