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由胆碱乙酰转移酶信使核糖核酸可变剪接变体编码的一种蛋白质优先定位于外周神经细胞和纤维中。

A protein encoded by an alternative splice variant of choline acetyltransferase mRNA is localized preferentially in peripheral nerve cells and fibers.

作者信息

Tooyama I, Kimura H

机构信息

Neuroanatomny Unit, Molecular Neuroscience Research Center, Shiga University of Medical Science, Otsu, Japan.

出版信息

J Chem Neuroanat. 2000 Jan;17(4):217-26. doi: 10.1016/s0891-0618(99)00043-5.

Abstract

Central cholinergic systems have been visualized by immunohistochemistry using antibodies to choline acetyltransferase (ChAT). Peripheral cholinergic cells and fibers, however, have been hardly detectable with most of these antibodies. This phenomenon suggests that a different form of ChAT may exist in peripheral tissues. Here we report two types of mRNA for ChAT expressed by alternative splicing in rat pterygopalatine ganglion. One is exactly identical with ChAT mRNA reported in the central nervous system (ChAT of a common type; cChAT). The other lacks exons 6, 7, 8 and 9, which was detected only in the pterygopalatine ganglion (ChAT of a peripheral type; pChAT). The peculiarity of pChAT in chemical structure, possessing a splice joint of the exons 5 and 10, led us to produce rabbit antisera against a recombinant peptide of 41 amino acids which spans over the splice joint. On Western blots using a successfully obtained antiserum, an intense band of about 50 kDa, corresponding to the expected molecular weight of pChAT, was detected in the pterygopalatine ganglion but not in the brain. Immunohistochemistry using the antiserum failed to reveal positive staining of known brain cholinergic structures, while it permitted us to observe peripheral, probably cholinergic, nerve cells and fibers including those in the pterygopalatine ganglion and enteric nervous system.

摘要

通过使用针对胆碱乙酰转移酶(ChAT)的抗体进行免疫组织化学,已可视化中枢胆碱能系统。然而,使用这些抗体中的大多数几乎检测不到外周胆碱能细胞和纤维。这种现象表明外周组织中可能存在不同形式的ChAT。在此,我们报告了大鼠翼腭神经节中通过可变剪接表达的两种ChAT mRNA类型。一种与中枢神经系统中报道的ChAT mRNA完全相同(常见类型的ChAT;cChAT)。另一种缺少外显子6、7、8和9,仅在翼腭神经节中检测到(外周类型的ChAT;pChAT)。pChAT在化学结构上的独特性,即具有外显子5和10的剪接接头,促使我们针对跨越该剪接接头的41个氨基酸的重组肽制备兔抗血清。在使用成功获得的抗血清进行的蛋白质印迹分析中,在翼腭神经节中检测到一条约50 kDa的强条带,与pChAT的预期分子量相对应,但在脑中未检测到。使用该抗血清进行免疫组织化学未能显示已知脑胆碱能结构的阳性染色,而它使我们能够观察到外周的、可能是胆碱能的神经细胞和纤维,包括翼腭神经节和肠神经系统中的那些。

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