Department of Experimental Genome Research, Research Institute for Microbial Diseases, Osaka University, Suita, Osaka, Japan.
Graduate School of Medicine, Osaka University, Suita, Osaka, Japan.
Biol Reprod. 2020 Aug 4;103(2):183-194. doi: 10.1093/biolre/ioaa083.
Developing a safe and effective male contraceptive remains a challenge in the field of medical science. Molecules that selectively target the male reproductive tract and whose targets are indispensable for male reproductive function serve among the best candidates for a novel non-hormonal male contraceptive method. To determine the function of these genes in vivo, mutant mice carrying disrupted testis- or epididymis-enriched genes were generated by zygote microinjection or electroporation of the CRISPR/Cas9 components. Male fecundity was determined by consecutively pairing knockout males with wild-type females and comparing the fecundity of wild-type controls. Phenotypic analyses of testis appearance and weight, testis and epididymis histology, and sperm movement were further carried out to examine any potential spermatogenic or sperm maturation defect in mutant males. In this study, we uncovered 13 testis- or epididymis-enriched evolutionarily conserved genes that are individually dispensable for male fertility in mice. Owing to their dispensable nature, it is not feasible to use these targets for the development of a male contraceptive.
开发安全有效的男性避孕药仍然是医学领域的一个挑战。选择性靶向男性生殖系统的分子，其靶标对于男性生殖功能是不可或缺的，这些是新型非激素男性避孕药方法的最佳候选物之一。为了确定这些基因在体内的功能，通过将 CRISPR/Cas9 成分的受精卵微注射或电穿孔，生成携带睾丸或附睾富集基因缺失的突变型小鼠。通过连续将敲除雄性与野生型雌性配对，并比较野生型对照的繁殖力，来确定雄性的繁殖力。进一步进行睾丸外观和重量、睾丸和附睾组织学以及精子运动的表型分析，以检查突变型雄性中是否存在任何潜在的精子发生或精子成熟缺陷。在这项研究中，我们揭示了 13 个睾丸或附睾富集的进化保守基因，这些基因在小鼠中单独对于雄性生育力是可有可无的。由于它们可有可无的性质，因此不可能将这些靶标用于开发男性避孕药。
