KIFC1 and myosin Va: two motors for acrosomal biogenesis and nuclear shaping during spermiogenesis of Portunus trituberculatus
KIFC1 and myosin Va: two motors for acrosomal biogenesis and nuclear shaping during spermiogenesis of Portunus trituberculatus
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KIFC1和肌球蛋白Va：三疣梭子蟹精子发生过程中顶体生物发生和核成形的两个马达
DOI：
10.1007/s00441-017-2638-4
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发表时间：
2017-09-01
影响因子：
3.6
通讯作者：
Yang, Wan-Xi
中科院分区：
文献类型：
--
作者：
Ma, Dan-Dan;Pan, Meng-Ying;Yang, Wan-Xi
为了探讨三疣梭子蟹精子发生的分子机制，通过快速扩增三疣梭子蟹睾丸cDNA的cDNA末端，克隆了全长运动蛋白KIFC 1和肌球蛋白Va，并使用各自的探针和特异性抗体来追踪它们在精子成熟过程中的定位。从基因序列设计反义探针，并用于检测每个基因的mRNA水平。荧光原位杂交（FISH）结果表明，kifc1和myosin Va的转录始于精子细胞中期，在顶体帽形成的部位kifc1 mRNA表达最活跃，myosin Va在顶体复合体中表达最集中。免疫荧光结果显示，KIFC 1和肌球蛋白Va在精子发生的各个阶段都有高表达。在早期精子细胞中，它们与细胞骨架一起随机分散在细胞质中。在中期，马达聚集在核的一侧上方，那里是后来形成顶体的地方。在晚期精子细胞和成熟精子中，KIFC 1紧密分布在核周区，表明其在核变形中起作用。肌球蛋白Va分布于顶体复合体，直至精子成熟。这表明肌球蛋白Va在顶体形成和成熟过程中的物质运输中的潜在作用。上述结果为KIFC 1和肌球蛋白Va在三疣梭子蟹精子发生中的重要作用提供了初步的证据。
To investigate the molecular mechanisms underlying the spermiogenesis of the swimming crab Portunus trituberculatus, full lengths of motor proteins KIFC1 and myosin Va were cloned by rapid-amplification of cDNA ends from P. trituberculatus testes cDNA, and their respective probes and specific antibodies were used to track their localization during sperm maturation. Antisense probes were designed from the gene sequences and used to detect the mRNA levels of each gene. According to the results of fluorescence in situ hybridization (FISH), the transcription of kifc1 and myosin Va began at the mid-stage of spermatids, with the kifc1 mRNA being most active at the location where the acrosome cap was formed and the myosin Va was more concentrated in the acrosome complex. Immunofluorescence results showed that KIFC1 and myosin Va were highly expressed in each stage of spermigenesis. In the early spermatids, they were randomly dispersed in the cytoplasm together with cytoskeletons. At the mid-stage, the motors were gathered above one side of the nucleus where the acrosome would later form. In the late spermatids and mature sperm, the KIFC1 was closely distributed in the perinuclear region, indicating its role in nucleus deformation. Myosin Va was distributed in the acrosome complex until sperm maturity. This suggests myosin Va's potential role in material transportation during acrosome formation and maturation. The above results provide a preliminary illustration of the essential roles of KIFC1 and myosin Va in the spermiogenesis of the swimming crab P. trituberculatus.