Characterization of a Novel Tektin Member, TEKT5, in Mouse Sperm
Characterization of a Novel Tektin Member, TEKT5, in Mouse Sperm
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DOI：
10.2164/jandrol.109.009456
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发表时间：
2011-01-01
影响因子：
--
通讯作者：
Gerton, George L.
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文献类型：
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作者：
Cao, Wenlei;Ijiri, Takashi W.;Gerton, George L.
Tektins是鞭毛的重要组成部分。小鼠tektins表达的改变或突变与精子活力缺陷相关，这是男性不育的一个原因。我们的鞭毛附属结构的蛋白质组学研究以前确定了一种新的tektin，TEKT 5，其功能是未知的。为了了解TEKT 5在小鼠精子中的作用，我们表征了小鼠Tekt 5基因的表达以及TEKT 5在生精细胞和精子中的存在。通过逆转录-聚合酶链反应（RT-PCR）和3 '-快速扩增cDNA末端，构建了编码Tekt 5转录本的完整cDNA，预测Tekt 5是一个62 730-道尔顿的蛋白质，具有一个不寻常的长C-末端。Tekt 5 mRNA在精子发生的晚期有高表达。在检测的组织中，Tekt 5 mRNA仅在睾丸和脑中表达，定量RT-PCR显示睾丸中Tekt 5 mRNA的表达量是脑中的6.8倍。在蛋白质水平上，TEKT 5存在于精子中，并在鞭毛的附属结构中富集。免疫荧光证实TEKT 5定位于整个精子尾部的鞭毛附属结构中。表达模式表明TEKT 5在精子发生过程中鞭毛形成中起重要作用，并与精子活力有关。
Tektins are important components of flagella. Alterations in the expression of or mutations in mouse tektins are correlated with defective sperm motility, a cause of male infertility. Our proteomic studies of flagellar accessory structures previously identified a novel tektin, TEKT5, whose function is unknown. To understand the role of TEKT5 in mouse sperm, we characterized the expression of the mouse Tekt5 gene and the presence of TEKT5 in spermatogenic cells and spermatozoa. A complete cDNA encoding the Tekt5 transcript was assembled following reverse transcription-polymerase chain reaction (RT-PCR) and 3'-rapid amplification of cDNA ends and predicted that TEKT5 is a 62 730-dalton protein with an unusual, long C-terminus. Tekt5 mRNA was highly expressed during late stages of spermiogenesis. Among examined tissues, Tekt5 mRNA was present only in testis and brain, and quantitative RT-PCR showed that the expression level of mRNA in testis was 6.8-fold higher than that in brain. At the protein level, TEKT5 was present in sperm and was enriched in the accessory structures of flagella. lmmunofluorescence confirmed that TEKT5 was localized throughout the sperm tail in flagellar accessory structures. The expression pattern suggests that TEKT5 plays an important role in flagella formation during spermiogenesis as well as being implicated in sperm motility.