A cholinergic chloride conductance in neurones of Helix aspersa

Inhibitory Cl- -mediated currents through cholinergic channels on the soma of identified neurones from the right parietal ganglion of Helix aspersa were studied under voltage clamp. Voltage-jump relaxation analysis showed that these currents decreased with hyperpolarization. In 3 microM-acetylcholine (ACh), the normalized fraction of channels in the open configuration (rho) decreased e-fold with each 191 mV of membrane hyperpolarization. The steady-state membrane conductance, G(infinity), decreased e-fold with each 128 mV of membrane hyperpolarization. The difference in the voltage sensitivities of rho and G(infinity) arose because of the voltage sensitivity of the instantaneous membrane conductance, G(0). G(0) rectified in the direction predicted by the Goldman-Hodgkin-Katz conductance model. The degree of rectification decreased when the internal Cl- concentration was raised. The relaxing currents were composed of two exponential components. At membrane potential (Vm) = -160 mV, 12 degrees C, the time constants of the two components were 4.1 ms and 21 ms in 3 microM-ACh, and 3.6 ms and 18 ms in 100 microM-tetramethylammonium (TMA). Fluctuation analysis in neurones loaded with Cl- yielded spectra which were composed of two Lorentzian components. In 3 microM-ACh the mean single-channel conductance (gamma) appeared to rise from a low value observed in cells with normal intracellular Cl- to 2.7 pS in cells whose internal Cl- concentration was raised four-fold. The voltage sensitivity of rho was attributed to the conformational change step of a gating mechanism having three kinetically distinguishable states.