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Department of Biology, University of Vermont, Burlington, VT 05405

Correspondence to Rona J. Delay: rdelay{at}uvm.edu

The vomeronasal organ (VNO) is an odor detection system that mediates many pheromone-sensitive behaviors. Vomeronasal sensory neurons (VSNs), located in the VNO, are the initial site of interaction with odors/pheromones. However, how an individual VSN transduces chemical signals into electrical signals is still unresolved. Here, we show that a Ca²⁺-activated Cl^– current contributes 80% of the response to urine in mouse VSNs. Using perforated patch clamp recordings with gramicidin, which leaves intracellular chloride undisrupted, we found that the urine-induced inward current (V_hold = –80 mV) was decreased in the presence of chloride channel blockers. This was confirmed using whole cell recordings and altering extracellular chloride to shift the reversal potential. Further, the urine-induced currents were eliminated when both extracellular Ca²⁺ and Na⁺ were removed. Using inside-out patches from dendritic tips, we recorded Ca²⁺-activated Cl^– channel activity. Several candidates for this Ca²⁺-activated Cl^– channel were detected in VNO by reverse transcription–polymerase chain reaction. In addition, a chloride cotransporter, Na⁺-K⁺-2Cl^– isoform 1, was detected and found to mediate much of the chloride accumulation in VSNs. Collectively, our data demonstrate that chloride acts as a major amplifier for signal transduction in mouse VSNs. This amplification would increase the responsiveness to pheromones or odorants.

Abbreviations: Ano, anoctamin; Best, bestrophin; DIDS, 4,4’-diisothiocyanatostilbene-2,2’-disulfonic acid; KCC, potassium-chloride cotransporter; MOE, main olfactory epithelium; NKCC, sodium-potassium-chloride cotransporter; NKCC1, Na⁺-K⁺-2Cl^– isoform 1; OSN, olfactory sensory neuron; RT, reverse transcription; TRPC2, transient receptor potential channel 2; VNO, vomeronasal organ; VSN, vomeronasal sensory neuron

© 2009 Yang and Delay
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