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首页> 外文期刊>The European Journal of Neuroscience >Molecular cloning and dendritic localization of rat SH3P7.
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Molecular cloning and dendritic localization of rat SH3P7.

机译：大鼠SH3P7的分子克隆和树突定位。

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摘要

SH3P7 was originally isolated by cloning SH3 domain ligand targets from a mouse embryo cDNA library. SH3P7 is an actin-binding protein implicated in antigen reception, JNK1 signalling, and Rac activation. It contains a drebrin homology sequence in its N-terminal region and a cortactin homology sequence (SH3 domain) in its C-terminal region. Both drebrin and cortactin are actin-binding proteins, and both have been suggested as possible regulators of the actin cytoskeleton in neurons. In the present study, we performed cDNA cloning of rat SH3P7, performed RT-PCR analysis, generated polyclonal antibodies against the recombinant rat SH3P7 protein, and examined the distribution of SH3P7 in the rat brain using immunohistochemistry. Sequence analysis revealed that there were at least four isoforms of the SH3P7 protein: SH3P7r1-SH3P7r4. RT-PCR analysis revealed that the predominant isoforms expressed in the brain were SH3P7r1 and SH3P7r3. The relative levels of isoform expression were similar among regions. Immunohistochemistry revealed that the most intense immunolabelling for SH3P7 was observed in the hippocampus and cerebellar cortex. Double-labelling studies with anti-SH3P7 antibody and other neuronal marker proteins revealed that SH3P7 was located primarily in dendrites, and in moderate amounts in cell bodies. Immunoreactivity was absent in the presynaptic terminals. In cultured astrocytes, SH3P7 was localized at protrusive structures of the cell periphery and in the cell body. We concluded that SH3P7 is ubiquitous in the rat brain, and occurs as several isoforms. Also, its dendritic localization suggests that SH3P7 is functionally linked to actin cytoskeleton organization in dendrites.
机译：SH3P7最初是通过从小鼠胚胎cDNA文库克隆SH3结构域配体靶标而分离的。 SH3P7是一种肌动蛋白结合蛋白，与抗原接收，JNK1信号传导和Rac激活有关。它在其N端区域包含drebrin同源序列，在其C端区域包含一个cortactin同源序列（SH3结构域）。 drebrin和cortactin都是肌动蛋白结合蛋白，都被认为可能是神经元中肌动蛋白细胞骨架的调节剂。在本研究中，我们进行了大鼠SH3P7的cDNA克隆，进行了RT-PCR分析，生成了针对重组大鼠SH3P7蛋白的多克隆抗体，并使用免疫组织化学检查了SH3P7在大鼠脑中的分布。序列分析表明，SH3P7蛋白至少有四个同工型：SH3P7r1-SH3P7r4。 RT-PCR分析表明，在大脑中表达的主要同工型为SH3P7r1和SH3P7r3。各区域间同工型表达的相对水平相似。免疫组织化学显示，在海马和小脑皮层中观察到了最强烈的SH3P7免疫标记。用抗SH3P7抗体和其他神经元标记蛋白进行的双标记研究表明，SH3P7主要位于树突中，在细胞体内的含量适中。突触前末端不存在免疫反应性。在培养的星形胶质细胞中，SH3P7定位在细胞外围和细胞体内的突出结构上。我们得出的结论是，SH3P7在大鼠脑中无处不在，并以几种同工型的形式出现。而且，其树突状定位表明SH3P7在功能上与树突中的肌动蛋白细胞骨架组织相关。

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