Background: Human sperm cryopreservation is a simple and effective approach for male fertility preservation. Methods: To identify potential proteomic changes in this process, data-independent acquisition (DIA), a technology with high quantitative accuracy and highly reproducible proteomics, was used to quantitatively characterize the proteomics of human sperm cryopreservation. Results: A total of 174 significantly differential proteins were identified between fresh and cryoperservated sperm: 98 proteins decreased and 76 proteins increased in the cryopreservation group. Bioinformatic analysis revealed that metabolic pathways play an important role in cryopreservation, including: propanoate metabolism, glyoxylate and dicarboxylate metabolism, glycolysis/gluconeogenesis, and pyruvate metabolism. Four different proteins involved in glycolysis were identified by Western blotting: GPI, LDHB, ADH5, and PGAM1. Conclusions: Our work will provide valuable information for future investigations and pathological studies involving sperm cryopreservation.

Introduction: Aging and delayed parenthood are major social concerns. Men older than 35 years, which is an advanced paternal age, experience reduced sperm quality and fertility.Methods: In this study, 12-month-old mice served as a model for males of advanced paternal age. RNA sequencing (RNA-seq) of epididymides from 2- and 12-month-old mice was performed.Results: Spermatogonia and sperm counts were significantly lower in these mice. We identified 449 differentially expressed genes by RNA-seq. Altered pathways were enriched using Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. Moreover, nine hub genes were identified from the DEGs, along with DEGs associated with mitochondria.Discussion: These results could enhance understanding of the molecular mechanisms underlying decreased male fertility in men of advanced paternal age and may aid in developing targeted treatment for male infertility related to aging.