Testis-expressed protein 33 is not essential for spermiogenesis and fertility in mice.
Testis-expressed protein 33 is not essential for spermiogenesis and fertility in mice.
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DOI：
10.3892/mmr.2021.11956
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发表时间：
2021-05
影响因子：
3.4
通讯作者：
Zheng Y
中科院分区：
文献类型：
--
作者：
Xia M;Xia J;Niu C;Zhong Y;Ge T;Ding Y;Zheng Y
基因表达分析显示，小鼠体内富含超过 2,300 个睾丸基因，而基因敲除模型表明，其中许多基因与雄性生育能力有关。然而，许多基因的功能尚未阐明。本研究的目的是鉴定小鼠睾丸表达蛋白33（TEX33）的表达模式，并探讨TEX33在雄性生殖中的作用。使用逆转录聚合酶链反应和蛋白质印迹测定来研究第一波精子发生过程中小鼠睾丸中 TEX33 的 mRNA 和蛋白质水平。还进行了免疫荧光分析，以确定 TEX33 蛋白在睾丸中的细胞和结构定位。 Tex33 基因敲除小鼠是通过 CRISPR/Cas9 基因编辑产生的。还进行了苏木精和伊红或高碘酸希夫 (PAS) 染色的组织学分析、计算机辅助精子分析 (CASA) 和生育力测试，以评估 TEX33 对小鼠精子发生和雄性生殖的影响。结果显示，Tex33 mRNA和蛋白仅在小鼠睾丸中表达，并在出生后21-28天（精子发生期）首次检测到；他们的表情一直持续到成年。免疫荧光分析显示，TEX33蛋白位于小鼠睾丸的精细胞和生精小管内的精子中，并在精子发生过程中特异性定位于顶体、鞭毛和manchette。这些结果表明 TEX33 可能在小鼠精子发生中发挥作用。然而，与野生型小鼠相比，Tex33 基因敲除小鼠的睾丸与体重比率没有表现出可检测到的差异。 PAS染色和CASA显示，缺乏Tex33的小鼠的精子发生和精子质量正常。此外，生育力检测表明Tex33基因敲除小鼠具有正常的生殖功能。总之，本研究的结果表明 TEX33 与精子发生有关，但对于精子发育和男性生育能力并不是必需的。
Gene expression analyses have revealed that there are >2,300 testis-enriched genes in mice, and gene knockout models have shown that a number of them are responsible for male fertility. However, the functions of numerous genes have yet to be clarified. The aim of the present study was to identify the expression pattern of testis-expressed protein 33 (TEX33) in mice and explore the role of TEX33 in male reproduction. Reverse transcription-polymerase chain reaction and western blot assays were used to investigate the mRNA and protein levels of TEX33 in mouse testes during the first wave of spermatogenesis. Immunofluorescence analysis was also performed to identify the cellular and structural localization of TEX33 protein in the testes. Tex33 knockout mice were generated by CRISPR/Cas9 gene-editing. Histological analysis with hematoxylin and eosin or periodic acid-Schiff (PAS) staining, computer-assisted sperm analysis (CASA) and fertility testing, were also carried out to evaluate the effect of TEX33 on mouse spermiogenesis and male reproduction. The results showed that Tex33 mRNA and protein were exclusively expressed in mouse testes and were first detected on postnatal days 21–28 (spermiogenesis phase); their expression then remained into adulthood. Immunofluorescence analysis revealed that TEX33 protein was located in the spermatids and sperm within the seminiferous tubules of the mouse testes, and exhibited specific localization to the acrosome, flagellum and manchette during spermiogenesis. These results suggested that TEX33 may play a role in mouse spermiogenesis. However, Tex33 knockout mice presented no detectable difference in testis-to-body weight ratios when compared with wild-type mice. PAS staining and CASA revealed that spermatogenesis and sperm quality were normal in mice lacking Tex33. In addition, fertility testing suggested that the Tex33 knockout mice had normal reproductive functions. In summary, the findings of the present study indicate that TEX33 is associated with spermiogenesis but is not essential for sperm development and male fertility.