Spatial Transcriptomic Profiling of Male Germ Cells from Control and Cbfb cKO mice

Much of the framework for spermatogenesis is established postnatally before sexual maturation. To evaluate the role of core-binding factor subunit β (CBFβ) in prepubertal male germline development, we generated male mice with conditional knockout of Cbfb using Blimp1-Cre (denoted Blimp1-Cbfb cKO). Outcomes from phenotype analysis revealed that Blimp1-Cbfb cKO males were completely infertile and experienced rapid germline degeneration prior to puberty. To evaluate the pathways downstream of Cbfb that coincide with this degeneration, we conducted spatial transcriptomics of heterozygous control and Blimp1-Cbfb cKO testes at postnatal days 6 (P6) and 21 (P21) and used fluorescence immunostaining to identify and capture cell-type specific transcriptomic signatures. Collectively, our analysis revealed central roles of CBFβ in maintenance, differentiation, and cell cycle checkpoint regulation within the prepubertal male germline.