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微管相关蛋白MAP2和tau在培养的大鼠脑少突胶质细胞中的表达。

Expression of microtubule-associated proteins MAP2 and tau in cultured rat brain oligodendrocytes.

作者信息

Müller R, Heinrich M, Heck S, Blohm D, Richter-Landsberg C

机构信息

Department of Biology, University of Bremen, D-28359 Bremen, Germany.

出版信息

Cell Tissue Res. 1997 May;288(2):239-49. doi: 10.1007/s004410050809.

Abstract

Oligodendrocytes in culture are characterized by large membranous sheets containing an elaborate network of microtubules. Microtubule-associated proteins (MAPs) participate in microtubule stability and the regulation of the cellular architecture. We have investigated the expression of two major groups of MAPs, MAP2 and tau, in cultured rat brain oligodendrocytes. Alternatively spliced isoforms of mRNAs encoding MAP2 and tau were assessed by means of reverse transcription and polymerase chain reaction using a newly designed set of MAP2- and tau-specific primers. The data were compared with data obtained with cultures of rat brain astrocytes and rat cerebral neurons, and adult rat brain. The results show that oligodendrocytes, similarly to neurons, express mainly MAP2c transcripts containing three microtubule-binding repeats. They also contain small amounts of MAP2b mRNA. Six low molecular weight tau isoforms, namely tau 1-6, have been described in the brain (Goedert et al. 1991). The major isoform of tau mRNA in oligodendrocytes was found to be tau 1, which represents a marker typical for immature neurons. Tau 2 and tau 4 isoforms were also detected, albeit at a very low level. Immunoblot analysis of oligodendroglia cell extracts confirmed the presence of tau protein. It migrates as a single polypeptide with an apparent molecular weight of approximately 55 kDa. In addition, oligodendrocytes express MAP2c protein, which migrates as a close double band with an apparent molecular weight around 70 kDa. Indirect immunofluorescence staining indicated that tau and MAP2 immunoreactivity was expressed in oligodendrocytes of immature and mature morphologies in the cell somata and cellular processes. Tau was particularly found in the end of the cellular extensions, and both proteins exhibited a distribution similar to myelin basic protein. Thus, oligodendroglia, like neuronal cells, contain microtubule-associated proteins, mainly MAP2c and the tau 1 isoform, although at a much lower level. The presence of these MAPs in myelin-forming cells further points to the functional significance of the cytoskeleton during oligodendrocyte differentiation, process outgrowth, and myelin formation.

摘要

培养的少突胶质细胞的特征是具有大的膜状薄片,其中包含精细的微管网络。微管相关蛋白(MAPs)参与微管稳定性和细胞结构的调节。我们研究了培养的大鼠脑少突胶质细胞中两类主要MAPs,即MAP2和tau的表达。通过使用新设计的一组MAP2和tau特异性引物进行逆转录和聚合酶链反应,评估了编码MAP2和tau的mRNA的可变剪接异构体。将数据与大鼠脑星形胶质细胞、大鼠脑神经元培养物以及成年大鼠脑获得的数据进行比较。结果表明,少突胶质细胞与神经元类似,主要表达含有三个微管结合重复序列的MAP2c转录本。它们还含有少量的MAP2b mRNA。大脑中已描述了六种低分子量tau异构体,即tau 1 - 6(Goedert等人,1991年)。少突胶质细胞中tau mRNA的主要异构体被发现是tau 1,它代表未成熟神经元的典型标志物。还检测到了tau 2和tau 4异构体,尽管水平非常低。少突胶质细胞提取物的免疫印迹分析证实了tau蛋白的存在。它作为一条单多肽迁移,表观分子量约为55 kDa。此外,少突胶质细胞表达MAP2c蛋白,它作为紧密的双条带迁移,表观分子量约为70 kDa。间接免疫荧光染色表明,tau和MAP2免疫反应性在未成熟和成熟形态的少突胶质细胞的细胞体和细胞突起中表达。特别在细胞延伸末端发现了tau蛋白,并且这两种蛋白都表现出与髓鞘碱性蛋白相似的分布。因此,少突胶质细胞与神经元细胞一样,含有微管相关蛋白,主要是MAP2c和tau 1异构体,尽管水平要低得多。这些MAPs在形成髓鞘的细胞中的存在进一步表明了细胞骨架在少突胶质细胞分化、突起生长和髓鞘形成过程中的功能意义。

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