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大鼠和猴小脑内GABAA/苯二氮䓬受体样免疫反应性

GABAA/benzodiazepine receptor-like immunoreactivity in rat and monkey cerebellum.

作者信息

Meinecke D L, Tallman J, Rakic P

机构信息

Department of Neuroanatomy, Yale University School of Medicine, New Haven, CT 06510.

出版信息

Brain Res. 1989 Jul 31;493(2):303-19. doi: 10.1016/0006-8993(89)91165-7.

Abstract

The cellular and subcellular localization of GABAA/benzodiazepine receptor-like immunoreactivity in the rat and monkey cerebellum has been studied with a monoclonal antibody (E9) directed against the alpha-subunit of purified GABAA/benzodiazepine receptors. At both the light and electron microscopic level E9 immunoreactivity is located in all 3 layers of the cerebellar cortex and within the deep cerebellar nuclei. The reaction product accumulates within the cytoplasm of neurons and their dendrites but axons are not immunoreactive. Glial cells in the white matter and the cortical layers are also unlabeled, although in some instances Bergmann glia do contain reaction product. The overall distribution and cellular and subcellular localization of E9 immunoreactivity is identical for both monkey and rat cerebellum. On the basis of cell size, morphology, and location it is evident that E9 immunoreactivity occurs in examples of all 5 neuronal types in the cerebellar cortex: Purkinje cells, Golgi type II cells, granule cells, and stellate and basket cells. However, the distribution of the reaction product within the cells is more selective. For example, electron microscopy demonstrates that axonal processes and terminals are not E9 immunoreactive with the single exception of the mossy fiber terminals in the granular layer. Also, examples of unlabeled axon terminals resembling those derived from Golgi type II cells, basket cells, and stellate cells form synapses with immunoreactive dendrites and cell bodies in the cortical layers. Finally, in the deep cerebellar nuclei unreactive axon terminals make symmetric synapses with immunostained neurons and dendrites. These results show that E9 monoclonal antibodies label neurons and portions of their processes which are postsynaptic in GABA-mediated inhibitory circuits, and demonstrates that this antiserum can be used as a morphological marker for cells which make GABAA/benzodiazepine receptors.

摘要

利用一种针对纯化的GABAA/苯二氮䓬受体α亚基的单克隆抗体(E9),研究了大鼠和猴小脑内GABAA/苯二氮䓬受体样免疫反应性的细胞和亚细胞定位。在光镜和电镜水平上,E9免疫反应性位于小脑皮质的所有三层以及小脑深部核团内。反应产物积聚在神经元及其树突的细胞质中,但轴突无免疫反应性。白质和皮质层中的神经胶质细胞也未被标记,尽管在某些情况下,伯格曼胶质细胞确实含有反应产物。E9免疫反应性在猴和大鼠小脑中的总体分布以及细胞和亚细胞定位是相同的。根据细胞大小、形态和位置,很明显E9免疫反应性出现在小脑皮质所有5种神经元类型中:浦肯野细胞、高尔基II型细胞、颗粒细胞以及星状细胞和篮状细胞。然而,反应产物在细胞内的分布更具选择性。例如,电子显微镜显示轴突过程和终末除颗粒层中的苔藓纤维终末外均无E9免疫反应性。此外,未标记的轴突终末,类似于源自高尔基II型细胞、篮状细胞和星状细胞的轴突终末,与皮质层中具有免疫反应性的树突和细胞体形成突触。最后,在小脑深部核团中,无反应性的轴突终末与免疫染色的神经元和树突形成对称突触。这些结果表明,E9单克隆抗体标记了在GABA介导的抑制性回路中作为突触后成分的神经元及其部分突起,并证明这种抗血清可作为具有GABAA/苯二氮䓬受体的细胞的形态学标记物。

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