Project description:We report the application of single-cell ATAC-seq to reconstruct the epigenetic program and differentiation trajectory associated with the retinoic acid (RA)-induced differentiation in cultured male germline stem cells (GSCs). We also identified key transcription factors and regulatory elements regulated by RA.
Project description:We report the application of single-cell ATAC-seq to reconstruct the epigenetic program and identify key transcription factors and cis-regulatory elements associated with the epithelial and mesenchymal heterogeneity and G9a inhibition in cultured male germline stem cells (GSCs). First, we identified epithelial-like and mesenchymal-like clusters in the sample. Pseudotime trajectory also indicated that GSCs could undergo EMT-like process with unique regulators in different stages. Lastly, we showed that G9a inhibition could suppress the EMT-like process in GSCs.
Project description:Establishing cell type–specific chromatin landscapes is essential for cellular identity, yet how these landscapes are generated and maintained remains poorly understood. Here, we demonstrate that the chromatin remodeler SMARCA5 facilitates epigenetic priming required for retinoic acid–induced differentiation in the male germline. Germ cell–specific deletion of Smarca5 results in a complete loss of differentiating spermatogonia, phenocopying vitamin A–deficient mice lacking retinoic acid signaling. During the perinatal transition from prospermatogonia to undifferentiated spermatogonia, SMARCA5 is recruited to DMRT1-binding sites located at distal putative enhancers and promoters of germline genes. The SMARCA5–DMRT1 pioneer complex establishes chromatin accessibility at these loci, generating poised enhancers and promoters that serve as retinoic acid receptor (RAR)–binding sites. Thus, SMARCA5 licenses transcriptional responses to retinoic acid that enable spermatogenic differentiation. Our findings uncover a critical epigenetic priming mechanism that links pioneer factor activity to external signal responsiveness in the germline.
Project description:Establishing cell type–specific chromatin landscapes is essential for cellular identity, yet how these landscapes are generated and maintained remains poorly understood. Here, we demonstrate that the chromatin remodeler SMARCA5 facilitates epigenetic priming required for retinoic acid–induced differentiation in the male germline. Germ cell–specific deletion of Smarca5 results in a complete loss of differentiating spermatogonia, phenocopying vitamin A–deficient mice lacking retinoic acid signaling. During the perinatal transition from prospermatogonia to undifferentiated spermatogonia, SMARCA5 is recruited to DMRT1-binding sites located at distal putative enhancers and promoters of germline genes. The SMARCA5–DMRT1 pioneer complex establishes chromatin accessibility at these loci, generating poised enhancers and promoters that serve as retinoic acid receptor (RAR)–binding sites. Thus, SMARCA5 licenses transcriptional responses to retinoic acid that enable spermatogenic differentiation. Our findings uncover a critical epigenetic priming mechanism that links pioneer factor activity to external signal responsiveness in the germline.
Project description:Establishing cell type–specific chromatin landscapes is essential for cellular identity, yet how these landscapes are generated and maintained remains poorly understood. Here, we demonstrate that the chromatin remodeler SMARCA5 facilitates epigenetic priming required for retinoic acid–induced differentiation in the male germline. Germ cell–specific deletion of Smarca5 results in a complete loss of differentiating spermatogonia, phenocopying vitamin A–deficient mice lacking retinoic acid signaling. During the perinatal transition from prospermatogonia to undifferentiated spermatogonia, SMARCA5 is recruited to DMRT1-binding sites located at distal putative enhancers and promoters of germline genes. The SMARCA5–DMRT1 pioneer complex establishes chromatin accessibility at these loci, generating poised enhancers and promoters that serve as retinoic acid receptor (RAR)–binding sites. Thus, SMARCA5 licenses transcriptional responses to retinoic acid that enable spermatogenic differentiation. Our findings uncover a critical epigenetic priming mechanism that links pioneer factor activity to external signal responsiveness in the germline.
Project description:mRNA expression of hearts obtained from mice with Stimulated by retinoic acid gene 6 (Stra6) germline deletion, Vitamin A deficiency (VAD) induced by lecithin-retinol acyltransferase (Lrat) germline deletion and feeding with vitamin A-deficient diet, and the combination of both. Mice were subjected to myocardial infarction (MI) or Sham surgery.