Laboratory for Reproductive Toxicology, INRS-Institut Armand-Frappier, Université du Québec, Laval, Quebec, Canada.
Department of Urology, Royal Victoria Hospital, McGill University, Montreal, Quebec, Canada.
Biol Reprod. 2020 Feb 12;102(1):92-101. doi: 10.1093/biolre/ioz171.
A crucial function of the epididymis is providing a surface glycocalyx that is important for sperm maturation and capacitation. Defensins are antimicrobial peptides expressed in the epididymis. In the macaque epididymis, defensin beta 126 (DEFB126) is important for sperm motility, however, it is not known whether this is the case in humans. The objectives were to determine: (1) if DEFB126 on human ejaculated sperm was correlated with sperm motility in fertile and infertile men, (2) that recombinant DEFB126 could induce immature sperm motility in vitro. Immunofluorescence staining indicated that the proportion of DEFB126-positive sperm was significantly higher in motile sperm. Furthermore, the proportion of DEFB126-labeled sperm was positively correlated with sperm motility and normal morphology. Additional studies indicated that the proportion of DEFB126-positive spermatozoa in fertile volunteers was significantly higher than in volunteers with varicocele, and in infertile volunteers with semen deficiencies. To determine the role of DEFB126 on sperm motility, the DEFB126 gene was cloned and used to generate recombinant DEFB126 in H9C2 cells (rat embryonic heart myoblast cells). Deletion mutations were created into two regions of the protein, which have been linked to male infertility. Immotile testicular spermatozoa were incubated with cells expressing the different forms of DEFB126. Full-length DEFB126 significantly increased motility of co-cultured spermatozoa. However, no increase in sperm motility was observed with the mutated forms of DEFB126. In conclusion, these results support the notion that DEFB126 is important in human sperm maturation and the potential use of DEFB126 for in vitro sperm maturation.
附睾的一个重要功能是提供表面糖萼,这对于精子成熟和获能很重要。防御素是在附睾中表达的抗菌肽。在猕猴的附睾中,防御素 β 126(DEFB126)对精子运动很重要,然而,在人类中是否也是如此还不得而知。本研究的目的是确定:(1)人精子中的 DEFB126 是否与生育力正常和不育男性的精子运动能力相关,(2)重组 DEFB126 是否可以在体外诱导不成熟精子运动。免疫荧光染色表明,在运动的精子中,DEFB126 阳性精子的比例明显更高。此外,DEFB126 标记精子的比例与精子运动和正常形态呈正相关。进一步的研究表明,在生育力正常的志愿者中,DEFB126 阳性精子的比例明显高于精索静脉曲张志愿者和精液缺乏的不育志愿者。为了确定 DEFB126 对精子运动的作用,克隆了 DEFB126 基因,并在 H9C2 细胞(大鼠胚胎心肌成肌细胞)中生成重组 DEFB126。对与男性不育相关的蛋白质的两个区域进行了缺失突变。将无活力的睾丸精子与表达不同形式的 DEFB126 的细胞共培养。全长 DEFB126 显著增加了共培养精子的运动能力。然而,突变形式的 DEFB126 并未观察到精子运动的增加。总之,这些结果支持了 DEFB126 对人类精子成熟很重要的观点,以及 DEFB126 在体外精子成熟中的潜在应用。