The testis-specific serine proteases PRSS44, PRSS46, and PRSS54 are dispensable for male mouse fertility
The testis-specific serine proteases PRSS44, PRSS46, and PRSS54 are dispensable for male mouse fertility
复制标题
DOI：
10.1093/biolre/ioz158
复制
发表时间：
2020-01-01
影响因子：
3.6
通讯作者：
Garcia, Thomas X.
中科院分区：
文献类型：
--
作者：
Holcomb, Richard J.;Oura, Seiya;Garcia, Thomas X.
高通量转录组学和蛋白质组学方法最近鉴定了大量生殖细胞特异性基因，其中许多基因仍有待通过功能遗传学方法进行研究。丝氨酸蛋白酶 (PRSS) 几乎占所有蛋白酶的三分之一，并且在我们的生物信息学筛选中，我们鉴定了许多睾丸特异性蛋白酶。在这项研究中，我们选择关注 Prss44、Prss46 和 Prss54，我们确认它们在小鼠和人类中具有睾丸特异性。根据对小鼠发育表达的分析，所有四种基因的表达仅限于精子发生的后期，同时在精子发生、精液形成或精子功能中具有潜在的功能作用。为了更好地了解雄性生殖需求和这些丝氨酸蛋白酶的功能作用，通过 CRISPR/Cas9 介导的 ES 细胞或合子方法单独消除每个基因。获得每个基因的纯合缺失突变体并分析表型变化。对睾丸重量、睾丸和附睾组织学、精子形态和生育能力的分析显示，与对照组相比，Prss44、Prss46 和 Prss54 敲除小鼠没有显着差异。因此，我们的结果证明这些基因并不是小鼠正常生育能力所必需的，尽管并不排除这些基因可能以冗余方式发挥作用的可能性。阐明这些新基因的个体功能需求或缺乏，对于更好地了解精子发生和精子成熟的潜在因素是必要的，这对于了解男性不育的病因和男性避孕药的开发具有重要意义。 CRISPR/Cas9系统。
High-throughput transcriptomics and proteomics approaches have recently identified a large number of germ cell-specific genes with many that remain to be studied through functional genetics approaches. Serine proteases (PRSS) constitute nearly one-third of all proteases, and, in our bioinformatics screens, we identified many that are testis specific. In this study, we chose to focus on Prss44, Prss46, and Prss54, which we confirmed as testis specific in mouse and human. Based on the analysis of developmental expression in the mouse, expression of all four genes is restricted to the late stage of spermatogenesis concomitant with a potential functional role in spermiogenesis, spermiation, or sperm function. To best understand the male reproductive requirement and functional roles of these serine proteases, each gene was individually ablated by CRISPR/Cas9-mediated ES cell or zygote approach. Homozygous deletion mutants for each gene were obtained and analyzed for phenotypic changes. Analyses of testis weights, testis and epididymis histology, sperm morphology, and fertility revealed no significant differences in Prss44, Prss46, and Prss54 knockout mice in comparison to controls. Our results thereby demonstrate that these genes are not required for normal fertility in mice, although do not preclude the possibility that these genes may function in a redundant manner. Elucidating the individual functional requirement or lack thereof of these novel genes is necessary to build a better understanding of the factors underlying spermatogenesis and sperm maturation, which has implications in understanding the etiology of male infertility and the development of male contraceptives.Summary sentenceThe testis-specific serine proteases Prss44, Prss46, and Prss54 are dispensable for male fertility based on phenotype analyses of knockout mice produced using the CRISPR/Cas9 system.