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脊髓切片中运动神经元的膜片钳研究:一种用于药物作用高分辨率分析的工具。

Patch clamp studies of motor neurons in spinal cord slices: a tool for high-resolution analysis of drug actions.

作者信息

Wang M Y, Kendig J J

机构信息

Department of Anesthesia, Stanford University School of Medicine, Stanford, CA 94305, USA.

出版信息

Acta Pharmacol Sin. 2000 Jun;21(6):507-15.

PMID:11360684
Abstract

AIM

To develop a tool for detailed analysis of spinally acting anesthetic and analgesic agents.

METHODS

Studies were done on visually identified motor neurons in 400 microns thick spinal cord slices from 14-23 d old rats using patch clamp techniques. Ethanol was used as a prototype general anesthetic agent.

RESULTS

Cell bodies in the ventrolateral horn identified as motor neurons by retrograde fluorescent labeling had a mean dimension of 32 +/- 5 microns (x +/- s, n = 25). Mean resting potential was -62.8 +/- 2.4 mV; input resistance was 44 +/- 24 M omega (n = 19). Threshold was -44 +/- 7 mV, and action potential amplitude 101 +/- 9 mV from baseline. Ethanol concentrations at and below 50-200 mmol/L decreased motor neuron excitability to the injected current; there was no effect on resting potential, but a variable reversible increase in input resistance. Ethanol reversibly depressed the excitatory postsynaptic potential, with a dose-response relationship similar to that previously observed for the population excitatory postsynaptic potential in intact spinal cord in vitro. Ethanol also reversibly depressed currents evoked by glutamate, reducing total charge transfer to 40% +/- 26% of control (x +/- s; n = 4).

CONCLUSION

Reduction of connectivity in this relatively thick slice preparation does not significantly modify drug actions. The actions of ethanol on excitatory synaptic transmission observed in intact spinal cord are in part due to postsynaptic effects on motor neurons.

摘要

目的

开发一种用于详细分析脊髓作用麻醉药和镇痛药的工具。

方法

使用膜片钳技术,对14 - 23日龄大鼠400微米厚脊髓切片中视觉识别的运动神经元进行研究。乙醇用作原型全身麻醉剂。

结果

通过逆行荧光标记鉴定为运动神经元的腹外侧角细胞体平均尺寸为32±5微米(x±s,n = 25)。平均静息电位为-62.8±2.4 mV;输入电阻为44±24 MΩ(n = 19)。阈值为-44±7 mV,动作电位幅度相对于基线为101±9 mV。50 - 200 mmol/L及以下的乙醇浓度降低了运动神经元对注入电流的兴奋性;对静息电位无影响,但输入电阻有可变的可逆增加。乙醇可逆性地抑制兴奋性突触后电位,剂量 - 反应关系与先前在完整脊髓体外观察到的群体兴奋性突触后电位相似。乙醇还可逆性地抑制谷氨酸诱发的电流,将总电荷转移减少至对照的40%±26%(x±s;n = 4)。

结论

在这种相对较厚的切片制备中连接性的降低不会显著改变药物作用。在完整脊髓中观察到的乙醇对兴奋性突触传递的作用部分归因于对运动神经元的突触后效应。

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