| Abstrakt: |
The effect of two commonly used sodium substitutes, tris and glucosamine, on the amplitude and kinetics of miniature end-plate currents (MEPCs), acetylcholine (ACh) induced end-plate currents (EPC) and EPC flucmations was studied in voltage clamped single muscle fibres from a monolayer preparation of the cutaneous pectoris muscle. Total replacement of sodium with each substitute shifted the reversal potential from -4.7 mV (normal sodium solution) to -36.6 mV(tris) and -49.0 mV (glucosamine). In tris and glucosamine substituted solutions the current (MEPC or EPC) — voltage relation became markedly nonlinear, with peak current decreasing with membrane hyperpolarization. Peak current at +40 mV, was unaltered in tris solutions and reduced in glucosamine substituted solutions. MEPCs decayed with a single exponential time course and the EPC fluctuation spectra were characterized by single Lorentzian functions in both normal sodium solution and each substituted solution. Analysis of EPC fluctuations demonstrated that both tris and glucosamine decrease single channel conductance and increase channel lifetime. Both effects were enhanced by either membrane hyperpolarization or by increasing the concentration of each substitute. In the presence of each cationic substitute, single channel conductance increased and mean channel lifetime decreased with membrane depolarization. Analysis of the data according to the constant field assumptions (Goldman, Hodgkin, Katz equation) provided an inadequate description of experimental currents obtained at hyperpolarized membrane potentials with total ion substitution. Shifts in reversal potential with partial substitution were, however, adequately predicted by the GHK equation. These results suggest that tris and glucosamine ions interact with end-plate channels to reduce cation permeability and decrease channel closing rates. The dependence of this block on the level of membrane potential suggests that these cations bind to site(s) within open end-plate channels. |
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