期刊文献

Identification of TSSK1 and TSSK2 as Novel Targets for Male Contraception 收藏

将TSSK1和TSSK2识别为男性避孕的新目标

原文求助发布源

作者

Saman Nayyab [1];Marıá Gracia Gervasi [2];Darya A. Tourzani [3];Yeva Shamailova [4];Hiroki Akizawa (https://orcid.org/0000-0003-1091-5629) [5];Mahboubeh Taghavi [6];Wei Cui (https://orcid.org/0000-0001-7281-3471) [7];Rafael Fissore [8];Ana Maria Salicioni [9];Gunda I. Georg (https://orcid.org/0000-0002-8900-9460) [10];Elizabeth Snyder (https://orcid.org/0000-0002-3786-3753) [11];Pablo E. Visconti (https://orcid.org/0000-0001-9320-7518) [12];

作者单位

[1]Department of Veterinary and Animal Sciences, University of Massachusetts-Amherst, Amherst, MA 01003, USA; [2]Department of Animal Science, University of Connecticut, Storrs, CT 06269, USA; [3]Department of Veterinary and Animal Sciences, University of Massachusetts-Amherst, Amherst, MA 01003, USA; [4]Department of Animal Science, Rutgers University, New Brunswick, NJ 08901, USA; [5]Department of Cell and Developmental Biology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; [6]Department of Veterinary and Animal Sciences, University of Massachusetts-Amherst, Amherst, MA 01003, USA; [7]Animal Models Core Facility, Institute for Applied Life Sciences (IALS), University of Massachusetts-Amherst, Amherst, MA 01003, USA; [8]Department of Veterinary and Animal Sciences, University of Massachusetts-Amherst, Amherst, MA 01003, USA; [9]Department of Veterinary and Animal Sciences, University of Massachusetts-Amherst, Amherst, MA 01003, USA; [10]Department of Medicinal Chemistry, Institute for Therapeutics Discovery and Development, University of Minnesota, Minneapolis, MN 55455, USA; [11]Department of Animal Science, Rutgers University, New Brunswick, NJ 08901, USA; [12]Department of Veterinary and Animal Sciences, University of Massachusetts-Amherst, Amherst, MA 01003, USA;

发布日期

2025-4-18

页码

601

DOI

10.3390/biom15040601

来源信息

Biomolecules ISSN：2218-273X, 2025年, 15卷, 4期, 601页

摘要

The testis-specific serine kinases (TSSKs) are post-meiotically expressed in testicular germ cells. Their testis-specific expression, together with their putative role in phosphorylation pathways, suggests that TSSKs have relevant roles in spermiogenesis, sperm function, or both. Independent Tssk3 and Tssk6 knockout mice, as well as the double Tssk1/Tssk2 KO males, are sterile. However, the double KO results are silent regarding the individual roles of TSSK1 and TSSK2. The aim of this study was to develop independent mutant mouse models of Tssk1 and Tssk2, using CRISPR/Cas9, to evaluate their independent roles in reproduction. Male heterozygous pups were used to establish one Tssk1 and two independent Tssk2 mutant lines. Natural mating mutant Tssk1 and Tssk2 homozygous males but not females were found to be sterile. Additionally, homozygous males have lower sperm numbers and decreased motility, and were infertile in vitro. Anti-TSSK2 antibodies were validated against Tssk2 mutants and used in Western blot and immunofluorescence experiments. TSSK2 is localized to the sperm head; importantly, it is present in the testes and sperm from Tssk1 mutant mice, confirming individual mutation. Our results indicate that both TSSK1 and TSSK2 are individually essential for male reproduction and support both kinases as suitable nonhormonal male contraceptive targets.

摘要译文

睾丸特异性丝氨酸激酶（TSSK）在睾丸生殖细胞中以丙酸后表达。他们的睾丸特异性表达以及它们在磷酸化途径中的推定作用表明，TSSK在精子发生，精子功能或两者兼而有之具有相关作用。独立的TSSK3和TSSK6基因敲除小鼠，以及双TSSK1/TSSK2 KO雄性，都是无菌的。但是，双重KO结果在TSSK1和TSSK2的个别角色上保持沉默。这项研究的目的是开发使用CRISPR/CAS9的TSSK1和TSSK2的独立突变小鼠模型，以评估其在繁殖中的独立作用。雄性杂合幼崽用于建立一个TSSK1和两个独立的TSSK2突变线。天然交配突变体TSSK1和TSSK2纯合雄性而不是女性无菌。此外，纯合雄性的精子数量较低，运动性降低，体外是不育的。对TSSK2突变体验证了抗TSSK2抗体，并用于蛋白质印迹和免疫荧光实验。TSSK2位于精子头上。重要的是，它存在于TSSK1突变小鼠的睾丸和精子中，证实了个体突变。我们的结果表明，TSSK1和TSSK2对于男性繁殖都是必不可少的，并且支持两个激酶作为合适的非激素男性避孕靶标。