Molecular cloning and dendritic localization of rat SH3P7
Molecular cloning and dendritic localization of rat SH3P7
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大鼠SH3P7的分子克隆和树突定位
DOI：
10.1046/j.0953-816x.2001.01727.x
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发表时间：
2001
影响因子：
3.4
通讯作者：
T. Shirao
中科院分区：
文献类型：
--
作者：
H. Yamazaki;Hideto Takahashi;Takeo Aoki;T. Shirao
SH3P7最初是通过从小鼠胚胎cDNA文库中克隆SH3结构域配体靶点分离得到的。SH3P7是一种与抗原接受、JNK1信号传导和Rac激活有关的肌动蛋白结合蛋白。它在N端含有一个drebrin同源序列，在C端含有一个SH3结构域。drebrin和cortinn都是肌动蛋白结合蛋白，两者都被认为是神经元肌动蛋白细胞骨架的可能调节因子。本研究对大鼠SH3P7进行了cDNA克隆，RT - PCR分析，制备了针对重组大鼠SH3P7蛋白的多克隆抗体，并利用免疫组织化学检测了SH3P7在大鼠脑内的分布。序列分析显示，SH3P7蛋白至少存在4种亚型：SH3P7r1-SH3P7r4。RT‐PCR分析显示，在大脑中表达的主要亚型是SH3P7r1和SH3P7r3。不同地区的相对表达水平相似。免疫组化结果显示，海马和小脑皮层对SH3P7的免疫标记最为强烈。抗SH3P7抗体和其他神经元标记蛋白的双标记研究表明，SH3P7主要位于树突中，在细胞体中也有少量存在。突触前终末无免疫反应性。在培养的星形胶质细胞中，SH3P7定位于细胞外周的突起结构和细胞体中。我们得出结论，SH3P7在大鼠大脑中普遍存在，并以几种亚型出现。此外，它的树突定位表明SH3P7在功能上与树突中的肌动蛋白细胞骨架组织有关。
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.
影响因子：
3.3
作者：
Kessels, MM;Engqvist-Goldstein, ÅEY;Drubin, DG
通讯作者：
Drubin, DG
影响因子：
3.3
作者：
Lappalainen, P;Kessels, MM;Drubin, DG
通讯作者：
Drubin, DG