The kinetics and mechanisms underlying the voltage dependence of inhibitory postsynaptic currents (IPSCs) recorded in the Mauthner cell (M cell) were investigated in the isolated medulla of 52-h-old zebrafish larvae, with the use of whole cell and outside-out patch-clamp recordings. 2. Spontaneous miniature IPSCs (mIPSCs) were recorded in the presence of 10(-6) M tetrodotoxin (TTX), 10 mM MgCl2, and 0.1 mM [CaCl2]o. Depolarizing the cell from -50 to +50 mV did not evoke any significant change in the distribution of mIPSC amplitudes, whereas synaptic currents were prolonged at positive voltages. The average decay time constant was increased twofold at +50 mV. 3. The voltage dependence of the kinetics of glycine-activated channels was first investigated during whole cell recording experiments. Currents evoked by voltage steps in the presence of glycine (50 microM) were compared with those obtained without glycine. The increase in chloride conductance (gCl-) evoked by glycine was time and voltage dependent. Inactivation and reactivation of the chloride current were observed during voltage pulses from 0 to -50 mV and from -50 to 0 mV, respectively, and they occurred with similar time constants (2-3 s). During glycine application, voltage-ramp analysis revealed a shift in the reversal potential (ECl-) occurring at all [Cl-]i tested. 4. The basis of the voltage sensitivity of glycine-evoked gCl- was first analyzed by measuring the relative changes in the total open probability (NPo) of glycine-activated channels with voltage.(ABSTRACT TRUNCATED AT 250 WORDS)
摘要
利用全细胞和外向膜片钳记录技术,在52小时龄斑马鱼幼虫的离体延髓中,研究了Mauthner细胞(M细胞)中记录的抑制性突触后电流(IPSC)电压依赖性的动力学和机制。2. 在存在10^(-6) M河豚毒素(TTX)、10 mM MgCl2和0.1 mM [CaCl2]o的情况下记录自发微小IPSC(mIPSC)。将细胞从-50 mV去极化到+50 mV不会引起mIPSC幅度分布的任何显著变化,而突触电流在正电压下会延长。在+50 mV时,平均衰减时间常数增加了两倍。3. 在全细胞记录实验中,首先研究了甘氨酸激活通道动力学的电压依赖性。将在存在甘氨酸(50 microM)时电压阶跃诱发的电流与没有甘氨酸时获得的电流进行比较。甘氨酸诱发的氯离子电导(gCl-)增加是时间和电压依赖性的。在从0到-50 mV和从-50到0 mV的电压脉冲期间,分别观察到氯离子电流的失活和再激活,并且它们以相似的时间常数(2-3 s)发生。在应用甘氨酸期间,电压斜坡分析显示在所有测试的[Cl-]i下反转电位(ECl-)发生了偏移。4. 首先通过测量甘氨酸激活通道的总开放概率(NPo)随电压的相对变化,分析了甘氨酸诱发的gCl-电压敏感性的基础。(摘要截断于250字)