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人脊髓培养物中的γ-氨基丁酸能神经元:正常及噻吩基苯环利定处理细胞的计算机辅助分析

GABAergic neurons in human spinal cord cultures: a computer-aided analysis of normal and thienyl-phencyclidine-treated cells.

作者信息

Calvet M C, Levallois C, Calvet J, Kamenka J M

机构信息

INSERM U336, Montpellier, France.

出版信息

Brain Res. 1993 Apr 16;608(2):299-309. doi: 10.1016/0006-8993(93)91471-4.

Abstract

gamma-Aminobutyric acid (GABA)-containing neurons were studied in dissociated cell cultures of human spinal cords from 6-10-week-old fetuses using immunohistochemistry with anti-GABA antibodies. Light microscopy showed two types of immunoreactive (IR) neurons: (1) IR neurons with short neuritic processes remaining near the cell body (small neuritic tree neurons); and (2) IR neurons with long neuritic processes extending far from the cell body (large neuritic tree neurons). Both types were studied at different ages in vitro, in control and in thienyl phencyclidine (TCP)-treated cultures by means of computer reconstructions and morphometric parameters. A discriminant analysis permitted the recognition of three populations: whatever the age, the control and TCP-treated neurons with small neuritic trees were not discriminated from each other and were considered to be one population whereas the 98 DIV control and both 21 DIV and 98 DIV TCP-treated cells with large neuritic trees were clearly separated from each other and from the small cell population. In all models, an astrocytic labeling, weaker than that of the neurons, was observed. The nature of these neurons (probably interneurons) intrinsic to the spinal cord is discussed in view of previous findings concerning the anatomical distribution and organization of the GABAergic system in the spinal cord.

摘要

利用抗γ-氨基丁酸(GABA)抗体进行免疫组织化学,对6至10周龄胎儿的人脊髓解离细胞培养物中的含GABA神经元进行了研究。光学显微镜显示出两种免疫反应性(IR)神经元:(1)具有短神经突起的IR神经元,其神经突起保留在细胞体附近(小神经树神经元);(2)具有长神经突起的IR神经元,其神经突起远离细胞体延伸(大神经树神经元)。通过计算机重建和形态计量学参数,在体外不同年龄、对照和噻吩基苯环利定(TCP)处理的培养物中对这两种类型的神经元进行了研究。判别分析识别出三个群体:无论年龄如何,具有小神经树的对照神经元和经TCP处理的神经元彼此之间没有区别,被视为一个群体,而具有大神经树的98天体外培养(DIV)对照神经元以及21 DIV和98 DIV经TCP处理的细胞与小细胞群体明显分开。在所有模型中,均观察到星形胶质细胞标记,其比神经元的标记弱。鉴于先前关于脊髓中GABA能系统的解剖分布和组织的研究结果,讨论了这些脊髓固有神经元(可能是中间神经元)的性质。

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