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升高的钾离子会增强中脑细胞培养物中发育的多巴胺能神经元对谷氨酸的易损性。

Elevated potassium enhances glutamate vulnerability of dopaminergic neurons developing in mesencephalic cell cultures.

作者信息

Andreeva N, Ungethum U, Heldt J, Marschhausen G, Altmann T, Andersson K, Gross J

机构信息

Brain Research Institute, Academy of Medical Sciences, Moscow, Russia.

出版信息

Exp Neurol. 1996 Feb;137(2):255-62. doi: 10.1006/exnr.1996.0024.

Abstract

This study examines the effects of high K+ concentration on the growth and development of mesencephalic cells and their glutamate vulnerability. Mesencephalic cell cultures obtained from Wistar rat embryos on the 14th gestational day were maintained for 14 days in medium with either normal (4.2 mM) or elevated (24.2 mM) potassium concentration. There was no significant difference due to various K+ concentration in cell growth and survival up to day in vitro (DIV) 13-14. In order to test the glutamate (Glu) vulnerability, cultures were treated with 100 mu M Glu for 15 min in salt solution on the DIV 3,6,8 and 13. Glu-induced neuronal damage was estimated 24 h later by measuring the neuron-specific enolase (NSE) content in the culture medium and by counting the number of tyrosine hydroxylase-immunoreactive (TH-IR) neurons. Glu had no damaging effect on the cells on DIV 3, but became pronounced beyond DIV 6. Elevated potassium concentration 24.2 mM) in the culture medium during development significantly increased neuronal vulnerability to Glu treatment, indicated by a higher increase of NSE content in the medium and by a more pronounced Glu-induced decrease of the number of TH-IR cells. The Glu-induced decrease of the number of TH-IR cells and of NSE-IR cells let us conclude that dopaminergic neurons are more vulnerable to glutamate than other neurons from mesencephalic culture.

摘要

本研究考察了高钾浓度对中脑细胞生长发育及其谷氨酸易损性的影响。从妊娠第14天的Wistar大鼠胚胎获取的中脑细胞培养物,在钾浓度正常(4.2 mM)或升高(24.2 mM)的培养基中维持14天。直到体外培养第13 - 14天,不同钾浓度对细胞生长和存活没有显著差异。为了测试谷氨酸(Glu)易损性,在体外培养第3、6、8和13天,将培养物在盐溶液中用100 μM Glu处理15分钟。24小时后,通过测量培养基中神经元特异性烯醇化酶(NSE)含量以及计数酪氨酸羟化酶免疫反应性(TH - IR)神经元数量来评估Glu诱导的神经元损伤。Glu在体外培养第3天对细胞没有损伤作用,但在第6天之后变得明显。发育期间培养基中升高的钾浓度(24.2 mM)显著增加了神经元对Glu处理的易损性,表现为培养基中NSE含量升高更多以及Glu诱导的TH - IR细胞数量减少更明显。Glu诱导的TH - IR细胞和NSE - IR细胞数量减少使我们得出结论,多巴胺能神经元比中脑培养物中的其他神经元对谷氨酸更易损。

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