Epididymis-specific RNase A family genes regulate fertility and small RNA processing

Joshua F Shaffer, Alka Gupta, Geetika Kharkwal, Edgardo E Linares, Andrew D Holmes, Upasna Sharma
{"title":"Epididymis-specific RNase A family genes regulate fertility and small RNA processing","authors":"Joshua F Shaffer, Alka Gupta, Geetika Kharkwal, Edgardo E Linares, Andrew D Holmes, Upasna Sharma","doi":"10.1101/2024.08.26.608813","DOIUrl":null,"url":null,"abstract":"Sperm small RNAs are implicated in intergenerational transmission of paternal environmental effects. Small RNAs generated by cleavage of tRNAs, known as tRNA fragments (tRFs), are an abundant class of RNAs in mature sperm, and can be modulated by environmental conditions. The ribonuclease(s) responsible for the biogenesis of tRFs in the male reproductive tract remains unknown. Angiogenin, a member of the Ribonuclease A superfamily (RNase A), cleaves tRNAs to generate tRFs in response to cellular stress. Four paralogs of Angiogenin, namely Rnase9, Rnase10, Rnase11, and Rnase12, are specifically expressed in the epididymis -a long, convoluted tubule where sperm mature and acquire fertility and motility. The biological functions of these genes remain largely unknown. Here, by generating mice deleted for all four genes (Rnase9-12-/-, termed KO for Knock Out), we report that these genes regulate fertility and RNA processing. KO mice showed complete male sterility. KO sperm fertilized oocytes in vitro but failed to efficiently fertilize oocytes in vivo, likely due to an inability of sperm to pass through the utero-tubular junction. Intriguingly, there were decreased levels of fragments of tRNAs (tRFs) and rRNAs (rRNA-derived small RNAs or rsRNAs) in the KO epididymis and epididymal luminal fluid, implying that Rnase9-12 regulate the biogenesis and/or stability of tRFs and rsRNAs. Importantly, KO sperm showed a dramatic decrease in the levels of tRFs, demonstrating a role of Rnase9-12 in regulating sperm RNA composition. Together, our results reveal an unexpected role of four epididymis-specific non-canonical RNase A family genes in fertility and RNA processing.","PeriodicalId":501246,"journal":{"name":"bioRxiv - Genetics","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2024-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"bioRxiv - Genetics","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1101/2024.08.26.608813","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0

Abstract

Sperm small RNAs are implicated in intergenerational transmission of paternal environmental effects. Small RNAs generated by cleavage of tRNAs, known as tRNA fragments (tRFs), are an abundant class of RNAs in mature sperm, and can be modulated by environmental conditions. The ribonuclease(s) responsible for the biogenesis of tRFs in the male reproductive tract remains unknown. Angiogenin, a member of the Ribonuclease A superfamily (RNase A), cleaves tRNAs to generate tRFs in response to cellular stress. Four paralogs of Angiogenin, namely Rnase9, Rnase10, Rnase11, and Rnase12, are specifically expressed in the epididymis -a long, convoluted tubule where sperm mature and acquire fertility and motility. The biological functions of these genes remain largely unknown. Here, by generating mice deleted for all four genes (Rnase9-12-/-, termed KO for Knock Out), we report that these genes regulate fertility and RNA processing. KO mice showed complete male sterility. KO sperm fertilized oocytes in vitro but failed to efficiently fertilize oocytes in vivo, likely due to an inability of sperm to pass through the utero-tubular junction. Intriguingly, there were decreased levels of fragments of tRNAs (tRFs) and rRNAs (rRNA-derived small RNAs or rsRNAs) in the KO epididymis and epididymal luminal fluid, implying that Rnase9-12 regulate the biogenesis and/or stability of tRFs and rsRNAs. Importantly, KO sperm showed a dramatic decrease in the levels of tRFs, demonstrating a role of Rnase9-12 in regulating sperm RNA composition. Together, our results reveal an unexpected role of four epididymis-specific non-canonical RNase A family genes in fertility and RNA processing.
附睾特异性 RNase A 家族基因调控生育能力和小 RNA 处理过程
精子小 RNA 与父代环境影响的代际传递有关。由 tRNA 分裂产生的小 RNA 被称为 tRNA 片段(tRNA fragments,tRFs),是成熟精子中含量丰富的一类 RNA,可受环境条件的影响。在男性生殖道中负责 tRFs 生物生成的核糖核酸酶仍然未知。Angiogenin是核糖核酸酶A超家族(RNase A)的成员,它能在细胞受压时裂解tRNA以生成tRFs。Angiogenin的四个旁系亲属,即Rnase9、Rnase10、Rnase11和Rnase12,在附睾中特异性表达,附睾是精子成熟、获得生育能力和运动能力的长而曲折的小管。这些基因的生物学功能在很大程度上仍然未知。在这里,我们通过产生全部四个基因都被删除的小鼠(Rnase9-12-/-,称为KO,意为 "敲除"),报告了这些基因对生育力和RNA处理的调控作用。KO 小鼠表现出完全雄性不育。KO精子在体外能使卵母细胞受精,但在体内却不能有效地使卵母细胞受精,这可能是由于精子无法通过子宫管交界处。耐人寻味的是,KO附睾和附睾管腔液中的tRNAs(tRFs)和rRNAs(rRNA衍生的小RNAs或rsRNAs)片段水平降低,这意味着Rnase9-12调节tRFs和rsRNAs的生物生成和/或稳定性。重要的是,KO精子中的tRFs水平急剧下降,这证明了Rnase9-12在调节精子RNA组成中的作用。总之,我们的研究结果揭示了四个附睾特异性非经典 RNase A 家族基因在生育和 RNA 处理中的意想不到的作用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
自引率
0.00%
发文量
0
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信