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加州海兔电压钳制完整神经细胞体中的钙尾电流。

Calcium tail currents in voltage-clamped intact nerve cell bodies of Aplysia californica.

作者信息

Eckert R, Ewald D

出版信息

J Physiol. 1983 Dec;345:533-48. doi: 10.1113/jphysiol.1983.sp014995.

Abstract

Calcium tail currents were measured in axotomized Aplysia neurones L2-L6 using a two-electrode voltage clamp and micro-electrodes of specially low resistance. Measurements were made at -40 mV following depolarizing pulses of 7 or 10 ms duration in the presence of 45 microM-tetrodotoxin and 200 mM-tetraethylammonium. Symmetrical currents were eliminated by addition of digitally stored current traces produced in response to equivalent hyperpolarizations. The remaining current, identified as a tail current, was blocked by replacement of extracellular calcium with cobalt or manganese. Computer fits showed that the tail current closely approximated the sum of two exponentially decaying components. The first had a time constant, tau 1, of 0.38 +/- 0.05 ms, which may have been frequency-limited by the speed of the clamp; the second had a time constant, tau 2, of 2.0 +/- 0.8 ms. A more slowly decaying third tail current component (tau 3 = 30 ms), which developed more slowly, may be related to the non-specific current rather than the calcium current. The tau 1 and tau 2 components of the tail current lost amplitude with increasing pulse duration along an approximately bi-exponential time course that resembled the time course of relaxation of the calcium current during a prolonged depolarization. The slow third component of the tail current showed no such inactivation. The amplitudes of the first and second components of the calcium tail current both increased as sigmoidal functions of the test pulse voltage, reaching half maximum at +20 mV and plateauing above +60 mV. The voltage dependencies of the two components were similar. The rate of decay of the tau 1 component increased with increasing temperature and with increasing negative potential, whereas tau 2 showed little dependence on these parameters. The rates of decay of neither the tau 1 nor the tau 2 component were affected by large changes in the amplitude of the test depolarization or in the amplitude of the tail current or by injection of calcium ions or EGTA. Thus, the kinetics of the tail current as resolved under our conditions appear to be virtually independent of calcium-mediated inactivation. Activation time constants (tau m) predicted from tau 1 are 3 to 5 times longer than the values of tau m determined from the half-time to peak of activation. These kinetics are slower than those reported for Limnaea by factors of 2.5 to 3.5.(ABSTRACT TRUNCATED AT 400 WORDS)

摘要

使用双电极电压钳和具有特别低电阻的微电极,在切断轴突的海兔神经元L2-L6中测量钙尾电流。在存在45微摩尔河豚毒素和200毫摩尔四乙铵的情况下,在持续7或10毫秒的去极化脉冲后,于-40毫伏下进行测量。通过添加响应等效超极化产生的数字存储电流迹线来消除对称电流。剩余的电流被鉴定为尾电流,用钴或锰替代细胞外钙可将其阻断。计算机拟合显示,尾电流非常接近两个指数衰减成分的总和。第一个成分的时间常数τ1为0.38±0.05毫秒,这可能受到钳制速度的频率限制;第二个成分的时间常数τ2为2.0±0.8毫秒。第三个衰减更慢的尾电流成分(τ3 = 30毫秒),其发展更慢,可能与非特异性电流而非钙电流有关。尾电流的τ1和τ2成分随着脉冲持续时间的增加,沿近似双指数时间进程失去幅度,这类似于长时间去极化期间钙电流的松弛时间进程。尾电流的缓慢第三成分没有这种失活现象。钙尾电流的第一和第二成分的幅度均作为测试脉冲电压的S形函数增加,在+20毫伏时达到最大值的一半,并在+60毫伏以上达到平稳。这两个成分的电压依赖性相似。τ1成分的衰减速率随温度升高和负电位增加而增加,而τ2对这些参数的依赖性很小。τ1和τ2成分的衰减速率均不受测试去极化幅度、尾电流幅度的大幅变化或钙离子或乙二醇双四乙酸(EGTA)注射的影响。因此,在我们的条件下解析出的尾电流动力学似乎实际上与钙介导的失活无关。由τ1预测的激活时间常数(τm)比根据激活的半峰时间确定的τm值长3至5倍。这些动力学比椎实螺报道的动力学慢2.5至3.5倍。(摘要截断于400字)

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