Loss of Cep135 causes oligoasthenoteratozoospermia and male infertility in mice

Centrosomal proteins (Cep), as crucial scaffolding molecules, play a pivotal role in the biogenesis of centrioles and the regulation of the cell cycle. To date, mutation in Cep135 has been reported to be closely associated with multiple morphological abnormalities of the flagella (MMAF) in humans. However, the specific mechanism of Cep135 in spermatogenesis and its detailed role in male infertility remains largely unexplored. In this study, we present compelling evidence that Cep135 functions as a pathogenic gene responsible for oligoasthenoteratozoospermia (OAT) and male infertility in mice. By selectively deleting Cep135 in premeiotic germ cells using Stra8-Cre mice crossed with Cep135^flox/flox mice, we observed that Cep135 knockdown produced abnormal sperm morphology, germ cell apoptosis and consequentlybecame complete infertility, but did not impact premeiosis. Scanning and transmission electron microscopy revealed defects in acrosome, flagellum, and head-to-tail connections during spermatogenesis. Proteomic analysis further indicated that CEP135 deletion led to a significant reduction in proteins mainly associated with acrosome formation, sperm heads, sperm flagellum and microtubule assembly. Additionally, CEP135 interacts with spermatogenic proteins SPATA6 and AKAP3, regulating their expression and stability. Deficiency in CEP135 or its interacting proteins resulted in ciliary shortening. In conclusion, our study profoundly unveils the central role of Cep135 in spermatogenesis and male fertility. This discovery not only deepens our comprehension of spermatogenesis but also furnishes a solid theoretical foundation and experimental evidence that can guide the formulation of therapeutic and preventive strategies for male infertility.