Human SRY Expression at the Sex-determining Period is Insufficient to Drive Testis Development in Mice

• Published in: Endocrinology (Philadelphia) 2022-01, Vol.163 (1), p.1
• Main Authors: Tsuji-Hosokawa, Atsumi, Ogawa, Yuya, Tsuchiya, Iku, Terao, Miho, Takada, Shuji
• Format: Article
• Language: English
• Subjects: Analysis; Animal experimentation; Animals; Anogenital; Compatibility; CRISPR; CRISPR-Cas Systems; Embryonic development; Embryos; Endocrinology; Exons; Female; Fetuses; Follicles; Gene Expression Regulation, Developmental; Genetic engineering; Genome editing; Genomes; Genomics; Genotype; Glutamine; Gonads; Gonads - metabolism; Humans; Male; Mice; Mice, Inbred C57BL; Mice, Transgenic; Molecular interactions; Nuclear Proteins - metabolism; Ovaries; Ovary - metabolism; Phenotype; Protein Domains; Regulatory sequences; Reproductive organs; Reproductive system; Sex; Sex determination; Sex Determination Processes; Sex Differentiation; Sex-Determining Region Y Protein - biosynthesis; Sex-Determining Region Y Protein - genetics; Signal Transduction; Sox9 protein; Testes; Testis - growth & development; Testis - metabolism; Testis - physiology; Transgenes; Transgenic mice; Y chromosomes
• ISSN: 0013-7227; 1945-7170
• DOI: 10.1210/endocr/bqab217

The sex-determining region of the Y chromosome, Sry/SRY, is an initiation factor for testis development in both humans and mice. Although the functional compatibility between murine SRY and human SRY was previously examined in transgenic mice, their equivalency remains inconclusive. Because molecular interaction and timeline of mammalian sex determination were mostly described in murine experiments, we generated a mouse model in which Sry was substituted with human SRY to verify the compatibility. The mouse model had the human SRY open reading frame at the locus of murine Sry exon 1-Sry(SRY) mice-and was generated using the CRISPR/Cas9 system. The reproductive system of the mice was analyzed. The expression of human SRY in the fetal gonadal ridge of Sry(SRY) mice was detected. The external and internal genitalia of adult Sry(SRY) mice were similar to those of wild-type females, without any significant difference in anogenital distance. Sry(SRY) mice obtained gonads, which were morphologically considered as ovaries. Histological analysis revealed that the cortical regions of gonads from adult Sry(SRY) mice contained few follicles. We successfully replaced genes on the Y chromosome with targeted genome editing using the CRISPR/Cas9 system. Because the Sry(SRY) XY mice did not develop testis, we concluded that human SRY was insufficient to drive testis development in mouse embryos. The difference in response elements and lack of glutamine-rich domains may have invalidated human SRY function in mice. Signal transduction between Sry/SRY expression and Sox9/SOX9 activation is possibly organized in a species-specific manner.