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首页> 外文期刊>Zoological Science >Immunolocalization of Spetex-1 at the Connecting Piece in Spermatozoa of the Musk Shrew (Suncus murinus)
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Immunolocalization of Spetex-1 at the Connecting Piece in Spermatozoa of the Musk Shrew (Suncus murinus)

机译：Spetex-1在麝香（Suncus murinus）精子连接部位的免疫定位

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摘要

Spetex-1, which has been isolated by differential display and rat cDNA library screening as a haploid spermatid-specific gene, encodes a protein with two coiled-coil motifs that locates at both the segmented column in the connecting piece and outer dense fibers-affiliated satellite fibrils in rat sperm flagella. Orthologs of Spetex-1 are identified in many animal species, including human, chimpanzee, macaque, cow, dog, African clawed frog, green spotted puffer, and zebrafish. In this study, we used RT-PCR in combination with 5' and 3' RACE (Rapid Amplification of cDNA End) technique to isolate Spetex-1 ortholog of the musk shrew (Suncus murinus), which yielded a full-length Suncus Spetex-1 gene containing an open reading frame of 1,908 base pairs encoding a protein of 636 amino acids with the predicted molecular mass of 72,348 Da. Suncus Spetex-1 has two coiled-coil motifs at 118184 and 242-276 amino acid residues, which is a characteristic shared by mammalian Spetex-1 proteins. To examine the subcellular localization of Spetex-1 in Suncus spermatozoa, we produced the anti-Suncus Spetex-1 antibody and carried out immunocytochemistry. In spite of that the primary structure of Suncus Spetex-1 is basically similar to that of rat and mouse Spetex-1, confocal laser scanning microscopy and immunoelectron microscopy revealed that Spetex-1 was restricted to the segmented column and capitulum in the connecting piece of Suncus spermatozoa and was not detected in other parts of flagella, suggesting a diversity of Spetex-1 localization in mammalian spermatozoa.
机译：Spetex-1已通过差异展示和大鼠cDNA文库筛选分离为单倍体精子特异性基因，该蛋白编码具有两个卷曲螺旋基序的蛋白质，该蛋白质位于连接片段中的分段柱和相关的外部密集纤维上大鼠精子鞭毛中的卫星原纤维。 Spetex-1的直系同源物可在许多动物物种中找到，包括人类，黑猩猩，猕猴，牛，狗，非洲爪蛙，绿斑河豚和斑马鱼。在这项研究中，我们将RT-PCR与5'和3'RACE（cDNA末端快速扩增）技术结合使用，分离了麝香sh（Suncus murinus）的Spetex-1直系同源物，从而产生了全长的Suncus Spetex- 1个基因，含有1,908个碱基对的开放阅读框，编码636个氨基酸的蛋白质，预测的分子量为72,348 Da。 Suncus Spetex-1在118184和242-276个氨基酸残基处有两个卷曲螺旋基序，这是哺乳动物Spetex-1蛋白共有的特征。为了检查Spetex-1在Suncus精子中的亚细胞定位，我们生产了抗Suncus Spetex-1抗体并进行了免疫细胞化学。尽管Suncus Spetex-1的主要结构与大鼠和小鼠Spetex-1的基本结构基本相同，但共聚焦激光扫描显微镜和免疫电子显微镜显示，Spetex-1限于连接段的分段柱和上皮。 Suncus spermatozoa，并在鞭毛的其他部位未检测到，提示哺乳动物精子中Spetex-1定位的多样性。

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