Identification of differentially expressed genes in human testis biopsies with defective spermatogenesis

Purpose: Sperm morphology and motility are major contributors to male-factor infertility, with many genes predicted to be involved. This study aimed to elucidate differentially expressed transcripts in human testis tissues of normal and abnormal spermatogenesis that could reveal new genes that may regulate sperm morphology and function.

Methods: Human testis biopsies were collected from men with well-characterized phenotypes of normal spermatogenesis, spermatid arrest, and Sertoli cell-only phenotype, and transcriptional differences were quantified by RNA-sequencing (RNA-Seq). Differentially expressed genes (DEGs) were filtered based on predominant expression in spermatids and gene functional annotations relevant to sperm morphology and motility. Selected 10 DEGs were validated by qRT-PCR and the localization of two proteins was determined in testis biopsies.

Results: The analysis revealed 6 genes (SPATA31E1, TEKT3, SLC9C1, PDE4A, CFAP47, and TNC) that are excellent candidates for novel genes enriched in developing human sperm. The immunohistochemical localization of two proteins, ORAI1 and SPATA31E1, in testis biopsies, verified that both are expressed in developing human germ cells, with SPATA31E1 enriched in late spermatocytes and spermatids.

Conclusion: This study identified human germ cell-enriched genes that could play functional roles in spermiogenesis and could thus be important in the development of morphologically normal, motile sperm.