Regulation of Spermatogenesis by Different Alternative Splicing Events of MEI1 in Mongolian Horse 

Abstract Background Mongolian horses are characterized by cold resistance, resistance to rough feeding, good endurance, strong disease resistance, and strong gregariousness. The normal production of sperm in male mammals forms the basis of reproduction and spermatogenesis and involves mitosis, meiosis, and metamorphosis of sperm cells. The MEI1 gene is involved in the meiosis cycle and is required for normal chromosome association during meiosis. Previous studies have shown that alternative splicing of MEI1 can promote spermatogenesis in Mongolian horses. In this paper, the regulatory effects of different MEI1 alternative splicing events on Mongolian horse spermatogenesis are investigated. Results In this study, two overexpressed lentiviral vectors with mutually exclusive exon (MXE) and skipped exon (SE) events of MEI1 were constructed and successfully used to infect Sertoli cells. After 72 h of viral infection, the expression of MEI1 was higher in the SE event than in the MXE event, as shown by fluorescence quantification; transcriptomics and metabolomics were then used to screen and annotate the differential genes and metabolites, and 193 differentially expressed genes (comprising 109 genes, such as MEI1, and 84 genes with upregulated and downregulated expression, respectively) and 11360 differentially expressed metabolites (comprising 7494 and 3866 metabolites with upregulated and downregulated expression, respectively) were screened. Differential genes and metabolites were mainly enriched in several metabolic pathways related to spermatogenesis. Differential genes such as IL31RA, ATP2B3, and CASQ2 were highly expressed in SE events, while IL11, PRLR, and CCR7 were highly expressed in MXE events. Metabolites such as folic acid and spermine were highly expressed during SE events, while citric acid and glutathione were highly expressed during MXE events. This suggests that both MXE and SE events of the MEI1 gene can promote the activity of the spermatogenesis signaling pathway. Conclusions The MXE and SE events of the MEI1 gene can promote the expression of spermatogenesis-related genes and metabolites, and these genes influence spermatogenesis by regulating the expression of metabolites. The results of this study provide a theoretical basis for exploring the regulatory mechanisms of alternative splicing events in spermatogenesis in Mongolian horses.