Background: While many testis-enriched genes have been identified as important regulators of the spermatogenic process, the specific roles played by several of these genes and their functional importance has yet to be fully clarified.
Methods: We employed a CRISPR/Cas9 approach to introduce a 5 bp in-frame deletion within the Spdye4a gene (Exon 2) of C57BL/6 mice (Spdye4a-/-). Fertility and sperm counts were evaluated. Testes tissues and cell suspensions were analyzed via histological and immunofluorescence staining. mRNA and protein levels of candidate genes were assessed through qPCR and Western blotting. In vitro fertilization was used to assess the ability of sperm cells to bind to egg cells.
Results: Spdye4a-/- mice did not exhibit any reduction in fertility, and exhibited comparable sperm counts, morphology and motility to those of wildtype littermates. Functionally, Spdye4a-/- sperm exhibited normal sperm-egg binding activity in vitro. Furthermore, the testes of Spdye4a-/- mice exhibited a full range of germ cells from spermatogonia to mature spermatozoa. No differences in the progression of meiotic prophase I were observed when comparing Spdye4a-/- and wildtype mice, indicating that the loss of Spdye4a had no adverse effect on spermatogenesis.
Discussion: Spdye4a is dispensable in the context of mice fertility and spermatogenesis. This study will prevent other laboratories from expending repeated efforts to generate similar knockout mice.
Keywords: Knockout; Male infertility; Mice; Spdye4a; Spermatogenesis.
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