Gene expression pattern of myosin Va during spermatogenesis of Chinese mitten crab, Eriocheir sinensis.
Gene expression pattern of myosin Va during spermatogenesis of Chinese mitten crab, Eriocheir sinensis.
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DOI：
10.1016/j.gene.2012.07.035
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发表时间：
2012-10
期刊：
影响因子：
3.5
通讯作者：
Xiao Sun;H. Mao;Wan-Xi Yang
中科院分区：
文献类型：
--
作者：
Xiao Sun;H. Mao;Wan-Xi Yang
肌球蛋白Va是一种F-肌动蛋白依赖的分子马达，具有多种功能，在小鼠精子发生过程中对顶体形成至关重要。螃蟹的精子有一个复杂的顶体，顶体周围有一个杯状的核。本研究采用简并PCR和RACE技术从中华绒螯蟹精巢中克隆了肌球蛋白Va的cDNA。肌球蛋白Va cDNA由125 bp的5′非翻译区（5′ UTR）、5331 bp的开放阅读框（ORF）和590 bp的3′ UTR组成。推测的肌球蛋白Va蛋白包含头部结构域、颈部结构域和尾部结构域。多重序列比对和系统进化树分析表明，E.中华肌球蛋白Va与脊椎动物肌球蛋白Va的关系比与无脊椎动物肌球蛋白V的关系更密切。中国人肌球蛋白Va在各种组织中均有表达。原位杂交结果表明，肌球蛋白Va mRNA在精子发生的整个过程中均有表达。实时荧光定量PCR结果表明，该基因在有丝分裂期和减数分裂期的表达水平高于精子发生期。综上所述，我们的工作表明肌球蛋白Va可能在E.中华精子发生
Myosin Va is an F-actin dependent molecular motor with multiple functions that are essential for acrosome formation in mouse spermiogenesis. The spermatozoon of the crab has a complicated acrosome surrounded by a cup-shaped nucleus. In the present study, the myosin Va cDNA was cloned from the testis of the Chinese mitten crab Eriocheir sinensis using degenerate PCR and rapid amplification of cDNA ends (RACE). The myosin Va cDNA consists of a 125bp 5′-untranslated region (5′ UTR), a 5331bp open reading frame (ORF) and a 590bp 3′ UTR. The putative myosin Va protein contains the head domain, neck domain and tail domain. Multiple alignment and phylogenetic tree showed that E. sinensis myosin Va is more closely related to the vertebrate myosin Va than to the invertebrate myosin V. E. sinensis myosin Va was expressed in various tissues. In situ hybridization demonstrated that myosin Va mRNA is located in the entire process of spermatogenesis. Quantitative real-time PCR indicated that the expression level at the mitotic and meiotic phases is higher than the spermiogenesis phase. Taken together, our work suggests that myosin Va may function in E. sinensis spermatogenesis.