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大鼠腹侧中脑器官型切片培养中多巴胺能和非多巴胺能神经元的电生理特性

Electrophysiological characterization of dopaminergic and non-dopaminergic neurones in organotypic slice cultures of the rat ventral mesencephalon.

作者信息

Steensen B H, Nedergaard S, Ostergaard K, Lambert J D

机构信息

Institute of Physiology, University of Aarhus, Denmark.

出版信息

Exp Brain Res. 1995;106(2):205-14. doi: 10.1007/BF00241116.

Abstract

The aim of the present study was to characterize electrophysiologically neurones in organotypic cultures of the rat ventral mesencephalon and to compare these results with results published for the same neurones in other types of preparation. Intracellular recordings were obtained in 3- to 8-week-old organotypic slice cultures of the ventral mesencephalon prepared from new-born rats. Dopaminergic neurones were distinguished from non-dopaminergic neurones by staining with the autofluorescent serotonin analogue 5,7-dihydroxytryptamine and briefly viewing the preparation with short exposures to ultraviolet (UV) light (365 nm). Short exposures to UV light did not affect the electrophysiological properties. There were no significant differences between dopaminergic and non-dopaminergic neurones with regard to resting membrane potential or action potential threshold and amplitude, and in both types of neurone spontaneous burst activity and glutamatergic excitatory postsynaptic potentials were seen. There were differences in the following parameters, which can be used to distinguish between the two types of neurone. Dopaminergic neurones had broad action potentials (2-9 ms), high input resistance (mean 81 M omega), were silent or fired spontaneously at a low frequency (0-9 Hz), and no spontaneous GABAA-ergic inhibitory postsynaptic potentials or inward rectification were present. In contrast, non-dopaminergic neurones had fast action potentials (0.6-3.2 ms), low input resistance (mean 32 M omega), were silent or fired spontaneously at relatively high firing frequency (0-28 Hz), and sometimes inhibitory postsynaptic potentials and inward rectification were seen. In the presence of 1 microM tetrodotoxin and 10 mM tetraethylammonium, Ca2+ spikes could be evoked in both dopaminergic and non-dopaminergic neurones. Dopaminergic neurones in 3- to 8-week-old organotypic slice cultures have a number of distinguishing electrophysiological characteristics similar to those recorded in other types of acute or cultured preparations. However, some intrinsic regulatory mechanisms, namely the slow oscillatory potentials, inward rectification and the K+ current, IA, seem to be missing in the cultured neurones.

摘要

本研究的目的是用电生理学方法对大鼠腹侧中脑器官型培养物中的神经元进行特性描述,并将这些结果与在其他类型制备物中对相同神经元所发表的结果进行比较。在由新生大鼠制备的3至8周龄腹侧中脑器官型切片培养物中进行细胞内记录。通过用自发荧光的血清素类似物5,7 - 二羟基色胺染色,并短暂暴露于紫外线(365nm)下观察制备物,将多巴胺能神经元与非多巴胺能神经元区分开来。短暂暴露于紫外线不会影响电生理特性。在静息膜电位、动作电位阈值和幅度方面,多巴胺能神经元与非多巴胺能神经元之间没有显著差异,并且在这两种类型的神经元中都观察到了自发爆发活动和谷氨酸能兴奋性突触后电位。在以下参数方面存在差异,这些差异可用于区分这两种类型的神经元。多巴胺能神经元具有宽动作电位(2 - 9毫秒)、高输入电阻(平均81MΩ),静息或低频(0 - 9Hz)自发放电,并且不存在自发的GABAA能抑制性突触后电位或内向整流。相比之下,非多巴胺能神经元具有快速动作电位(0.6 - 3.2毫秒)、低输入电阻(平均32MΩ),静息或相对高频(0 - 28Hz)自发放电,并且有时可见抑制性突触后电位和内向整流。在存在1μM河豚毒素和10mM四乙铵的情况下,多巴胺能和非多巴胺能神经元均可诱发Ca2 + 尖峰。3至8周龄器官型切片培养物中的多巴胺能神经元具有许多独特的电生理特征,与在其他类型的急性或培养制备物中记录的特征相似。然而,一些内在调节机制,即慢振荡电位、内向整流和钾电流IA,在培养的神经元中似乎缺失。

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