Project description:We report bulk RNA-sequencing for synchronized adult Sertoli cells in germ cell-sufficient (RFPxAMH-Cre) and germ cell-deficient (NANOS2 KO) mice. We also have single-cell RNA-sequencing data for RFPxAMH-Cre sorted Sertoli cells from unsynchronized adult testes (not included in this series).
Project description:Sertoli cells accumulate glycogen within a specific developmental window, peaking at embryonic day (E) 12.5 and rapidly depleting by E13.5. Using immunofluorescence and single-nuclei RNA-seq, we identified the regulatory mechanisms underlying this transient glycogen storage. Glycogen accumulation correlates with elevated PPP1R3C expression, while its degradation coincides with increased glycogen phosphorylase (PYGB) levels. Functional assays revealed that glycogen metabolism is not required for Sertoli cell differentiation but is essential for germ cell survival. Pharmacological inhibition of glycogen breakdown resulted in germ cell loss, which could not be rescued by exogenous lactate or glucose, highlighting a critical metabolic dependency. Glycogen-derived lactate is transported via the MCT4/MCT1 shuttle from Sertoli cells to germ cells, supporting their survival. These findings establish glycogen metabolism as a key determinant of germ cell viability, emphasizing a metabolic coupling between Sertoli and germ cells during testis development with implications for reproductive health.
Project description:Non-obstructive azoospermia (NOA) is a highly heterogeneous and complex disease. However, there is lack of the research on latent biological mechanisms. Here, we performed single-cell RNA sequencing (scRNA-seq) and Assay for Transposase-Accessible Chromatin using sequencing (scATAC-seq) on patients with obstructive azoospermia and NOA. Our study identified 12 germ cell subtypes and 8 sertoli cell subtypes. Further analysis identified several stage-specific marker genes of germ cells, such as ID4, TEX19, SCML1, and DMC1 and identified several stage-specific marker genes of sertoli cells, such as SOX2, BEND2, MLC1, PGAM2, and CFH. Furthermore, Notch1/2/3 signaling and integrin were involved in the interaction between germ cells and sertoli cells. Overall, we mapped the scRNA-seq and scATAC-seq of human spermatogenesis, and analyzed germ cell and somatic cell types and their key molecular markers. It gives us a comprehensive insight in spermatogenesis and provide a fresh perspective on diagnosis and treatment for NOA.
Project description:Sertoli cells accumulate glycogen within a specific developmental window, peaking at embryonic day (E) 12.5 and rapidly depleting by E13.5. Using immunofluorescence and single-nuclei RNA-seq, we identified the regulatory mechanisms underlying this transient glycogen storage. Glycogen accumulation correlates with elevated PPP1R3C expression, while its degradation coincides with increased glycogen phosphorylase (PYGB) levels. Functional assays revealed that glycogen metabolism is not required for Sertoli cell differentiation but is essential for germ cell survival. Pharmacological inhibition of glycogen breakdown resulted in germ cell loss, which could not be rescued by exogenous lactate or glucose, highlighting a critical metabolic dependency. Glycogen-derived lactate is transported via the MCT4/MCT1 shuttle from Sertoli cells to germ cells, supporting their survival. These findings establish glycogen metabolism as a key determinant of germ cell viability, emphasizing a metabolic coupling between Sertoli and germ cells during testis development with implications for reproductive health.