School of Life Science and Technology, Tong Ji University, Shanghai, China.
State Key Laboratory of Molecular Biology, Shanghai Key Laboratory of Molecular Andrology, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology (SIBCB), Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai, China.
FASEB J. 2018 Mar;32(3):1354-1363. doi: 10.1096/fj.201700936R. Epub 2018 Jan 3.
The epididymis is a male reproductive organ involved in posttesticular sperm maturation and storage, but the mechanism underlying sperm maturation remains unclear. β-Defensins (Defbs) belong to a family of small, cysteine-rich, cationic peptides that are antimicrobial and modulate the immune response. A large number of Defb genes are expressed abundantly in the male reproductive tract, especially in the epididymis. We and other groups have shown the involvement of several Defb genes in regulation of sperm function. In this study, we found that Defb23, Defb26, and Defb42 were highly expressed in specific regions of the epididymis. Rats with CRISPR/Cas9-mediated single-gene disruption of Defb23, Defb26, or Defb42 had no obvious fertility phenotypes. Those with the deletion of Defb23/ 26 or Defb23/ 26/ 42 became subfertile, and sperm isolated from the epididymal cauda of multiple-mutant rats were demonstrated decreased motility. Meanwhile, the sperm showed precocious capacitation and increased spontaneous acrosome reaction. Consistent with premature capacitation and acrosome reaction, sperm from multiple-gene-knockout rats had significantly increased intracellular calcium. These results suggest that Defb family members affect sperm maturation by a synergistic pattern in the epididymis.-Zhang, C., Zhou, Y., Xie, S., Yin, Q., Tang, C., Ni, Z., Fei, J., Zhang, Y. CRISPR/Cas9-mediated genome editing reveals the synergistic effects of β-defensin family members on sperm maturation in rat epididymis.
附睾是男性生殖器官的一部分，参与睾丸后精子成熟和储存，但精子成熟的机制尚不清楚。β-防御素（Defbs）属于富含半胱氨酸的阳离子小肽家族，具有抗菌和调节免疫反应的作用。大量 Defb 基因在男性生殖道中大量表达，尤其是在附睾中。我们和其他研究小组已经表明，一些 Defb 基因参与了精子功能的调节。在这项研究中，我们发现 Defb23、Defb26 和 Defb42 在附睾的特定区域高度表达。使用 CRISPR/Cas9 介导的单个基因敲除大鼠的 Defb23、Defb26 或 Defb42 ，没有明显的生育表型。那些缺失 Defb23/26 或 Defb23/26/42 的大鼠变得亚不育，从多个突变大鼠的附睾尾部分离出的精子显示运动能力下降。同时，精子显示出过早的获能和增加的自发顶体反应。与过早获能和顶体反应一致，来自多基因敲除大鼠的精子具有显著增加的细胞内钙。这些结果表明，Defb 家族成员通过协同模式影响附睾中的精子成熟。
