Nicotinic acetylcholine sensitivity of rat petrosal sensory neurons in dissociated cell culture
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Using whole-cell, patch-clamp techniques we investigated acetylcholine (ACh) sensitivity of dissociated sensory neurons from rat petrosal ganglia after 4 h-14 days in vitro. In approx. 68% of petrosal neurons (PN; n = 109) ACh, applied by fast perfusion or pressure ejection from a 'puffer' pipette, caused a rapid depolarization associated with a conductance increase. Under voltage clamp near the resting potential (approx. - 60 mV), ACh induced a hexamethonium-sensitive, inward current (IACh), mimicked by nicotine application, suggesting the presence of neuronal nicotinic acetylcholine receptors (nAChR). The reversal potential of IACh occurred near 0 mV (n = 4), a region where the I-V curve displayed a prominent rectification. The dose-response relation for IACh versus ACh concentration was fitted by the Hill equation with EC50 = approx. 33.9 microM and Hill coefficient = approx. 1.6. The activation phase of IACh was well fitted by a single exponential with mean (+/- S.E.M.) time constant of 102 +/- 82 ms (n = 6); the desensitization phase of IACh was best fitted by the sum of two exponentials, with time constant of 870 +/- 210 ms (n = 6) and 8576 +/- 1435 ms (at -70 mV). Fluctuation analysis yielded an apparent single-channel conductance of 21.6 +/- 10 pS (mean +/- S.E.M.; n = 4). These data indicate that a major subpopulation of sensory neurons in visceral petrosal ganglia of the rat express nAChR. Thus, if similar receptors are present on corresponding nerve terminals, they could mediate fast afferent excitation in response to ACh released at peripheral targets, e.g., the chemosensory carotid body.
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