Rln3a is a prerequisite for spermatogenesis and fertility in male fish
Rln3a is a prerequisite for spermatogenesis and fertility in male fish
复制标题
Rln3a是雄性鱼类精子发生和生育能力的先决条件
DOI：
10.1016/j.jsbmb.2019.105517
复制
发表时间：
2020-03
影响因子：
4.1
通讯作者：
Zhou Linyan
中科院分区：
文献类型：
--
作者：
Yang Lanying;Li Yanlong;Wu You;Sun Shaohua;Song Qiang;Wei Jing;Sun Lina;Li Minghui;Wang Deshou;Zhou Linyan
Relaxin3(RLN3)在能量平衡中的重要作用已经得到了很好的研究，但在哺乳动物中RLN3调节生殖的机制仍然不清楚。尽管在几种硬骨鱼中已经发现了类似物的特征，但它们的功能在很大程度上仍不清楚。在本研究中，从尼罗罗非鱼(Oreochromis Niloticus)的精巢中鉴定出两个类似的rln3基因，分别为rln3a和rln3b，其中rln3a主要在睾丸中表达，而rln3b在脑中表达最丰富。原位杂交结果表明，rln3aA在睾丸间质细胞中大量表达。为了解Rln3基因在睾丸发育中的作用，利用CRISPR/Cas9技术构建了同源的空rln3基因突变系。形态学观察表明，rln3agene零突变导致睾丸肥大，GSI显著增加。但在精原细胞、精母细胞、精子细胞和精子等不同时期的生精细胞明显减少。沉默rln3agene抑制了生殖细胞和间质细胞中关键基因的表达。Rln3a缺乏导致11-KT生成显著减少，可能通过负反馈的方式刺激垂体FSH和LH的生成上调。XY鱼rnn3a基因突变导致性腺功能低下，精子畸形，精子活力显著下降，表明与野生型雌性−/−XY鱼杂交所获得的后代死亡率较高。有趣的是，重组人RLN3注射显著提高了rIn3a−/−XY鱼的精子活力。此外，hCG处理还刺激了XY鱼类固醇合成酶基因的表达和11-KT的产生，而这些基因和11-KT的产生被rIn3抑制。综上所述，本研究首次利用基因敲除模型证明了Rln3a是通过HPG轴在睾丸中产生雄激素所必需的介质，并且在鱼类的精子发生、精子运动和雄性生育中起着重要作用。
The essential roles of Relaxin3 (RLN3) in energy homeostasis had been well investigated, while the mechanisms of RLN3 regulating reproduction remain to be elusive in mammals. Although tworln3paralogues have been characterized in several teleosts, their functions still remain largely unknown. In this study, two paralogousrln3genes, represented asrln3aandrln3b, were identified from the testis of Nile tilapia (Oreochromis niloticus).Rln3awas dominantly expressed in testis, while the most abundantrln3bexpression was in brain. In situ hybridization demonstrated thatrln3ais abundantly expressed in the Leydig cells of the testis. To understand the role of Rln3 in the testicular development, homologous null-rln3agene mutant line was constructed by CRISPR/Cas9 technology. Morphological observation demonstrated that null mutation ofrln3agene caused testicular hypertrophy and a significant increase of GSI. However, a significant decrease of spermatogenic cells at different phases, i.e. spermatogonia, spermatocytes, spermatids and sperms was found. Silencing ofrln3agene repressed the expression of key genes in germ cell and Leydig cell. Deficiency of Rln3a led to the significant decrease of 11-KT production, which stimulated the up-regulation of both FSH and LH production in the pituitary via a negative feedback manner possibly. Mutation ofrln3ain XY fish led to the hypogonadism with sperm deformation, significant decrease of fertility, and sperm motility, revealing as the high mortality of the offspring obtained by crossing the wild type female andrln3a−/−XY fish. Interestingly, recombinant human RLN3 injection significantly enhanced the sperm motility inrln3a−/−XY fish. Moreover, hCG treatment stimulated the expression of steroidogenic enzyme genes and 11-KT production, which were repressed byrln3amutation in XY fish. Taken together, this study, for the first time by using a gene knockout model, proved that Rln3a is an indispensable mediator for androgen production in testis via HPG axis, and plays an essential role in spermatogenesis, sperm motility and male fertility in fish.
登录
查看更多内容
影响因子：
4.8
作者：
Christophe Lenglos;J. Calvez;E. Timofeeva
通讯作者：
Christophe Lenglos;J. Calvez;E. Timofeeva
影响因子：
6.7
作者：
Kohn TP;Louis MR;Pickett SM;Lindgren MC;Kohn JR;Pastuszak AW;Lipshultz LI
通讯作者：
Lipshultz LI
影响因子：
56.9
作者：
Bowles, J;Knight, D;Koopman, P
通讯作者：
Koopman, P
影响因子：
2.7
作者：
Kusakabe, Makoto;Ishikawa, Asano;Kitano, Jun
通讯作者：
Kitano, Jun
影响因子：
4.8
作者：
Park, Jae-Il;Semyonov, Jenia;Hsu, Sheau Yu Teddy
通讯作者：
Hsu, Sheau Yu Teddy