Identification of TMCO2 as an acrosome-associated protein during rat spermiogenesis.,Molecular Reproduction and Development

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Identification of TMCO2 as an acrosome-associated protein during rat spermiogenesis.
Molecular Reproduction and Development ( IF 2.7 ) Pub Date : 2020-06-27 , DOI: 10.1002/mrd.23396
Takane Kaneko ₁ , Saori Toh ₁ , Izumi Mochida ₁ , Naoki Iwamori ₁ , Tetsuichiro Inai ₂ , Hiroshi Iida ₁

Affiliation

We isolated the transmembrane and coiled‐coil domains 2 (Tmco2 ) gene using a polymerase chain reaction‐based subtraction technique. Tmco2 is predominantly expressed in rat testes starting from 4 weeks of age. Rat TMCO2 consists of 187 amino acids with a predicted molecular mass of 20.6 kDa. When expressed in COS7 cells, TMCO2 was found as vesicle‐like structures in the cytoplasm, whereas TMCO2ΔTM lacking the transmembrane (TM) region was found diffused in the cytoplasm. These results suggest that the TM region in TMCO2 is essential for its specificity of localization. Immunocytochemical analyzes indicated that rat TMCO2 was localized as small semiluminate bodies or cap‐like structures in the vicinity of round spermatid nuclei and as curved lines associated with nuclei of elongated spermatids and caput epididymal spermatozoa. However, it was detected in only a small part of cauda epididymal spermatozoa. Double immunolabeling of the spermatids and spermatozoa with the anti‐TMCO2 antibody and the monoclonal anti‐MN7 antibody showed that TMCO2 was predominantly associated with the inner acrosomal membrane in spermatids and caput epididymal spermatozoa. Our findings suggest that TMCO2 might be involved in the process of acrosome biogenesis, especially binding of acrosome to a nucleus, during spermiogenesis.

中文翻译：

TMCO2 鉴定为大鼠精子发生过程中的顶体相关蛋白。

我们使用基于聚合酶链反应的消减技术分离了跨膜和卷曲螺旋结构域 2 ( Tmco2 ) 基因。从 4 周龄开始，Tmco2主要在大鼠睾丸中表达。大鼠 TMCO2 由 187 个氨基酸组成，预计分子量为 20.6 kDa。当在 COS7 细胞中表达时，TMCO2 在细胞质中呈囊泡样结构，而缺乏跨膜 (TM) 区域的 TMCO2ΔTM 在细胞质中扩散。这些结果表明 TMCO2 中的 TM 区域对于其定位特异性至关重要。免疫细胞化学分析表明，大鼠 TMCO2 定位为圆形精子细胞核附近的小半发光体或帽状结构，以及与细长精子细胞和带头附睾精子的细胞核相关的曲线。然而，仅在一小部分附睾尾精子中检测到。用抗TMCO2抗体和单克隆抗MN7抗体对精子和精子进行双重免疫标记表明，TMCO2主要与精子细胞和附睾精子的顶体内膜相关。我们的研究结果表明，TMCO2 可能参与精子发生过程中顶体生物发生过程，特别是顶体与细胞核的结合。

更新日期：2020-07-28

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