biostudies-literatureNagano MExploratory Research for Advanced TechnologyNIAID NIH HHSNINDS NIH HHSOpen Philanthropy ProjectHFSPNCI NIH HHSNational Institutes of HealthJapan Society for the Promotion of Sciencee110600https://www.ebi.ac.uk/biostudies/studies/S-EPMC9251848biostudies-literatureUnknown41(13)Germ cells are unique in engendering totipotency, yet the mechanisms underlying this capacity remain elusive. Here, we perform comprehensive and in-depth nucleome analysis of mouse germ-cell development in vitro, encompassing pluripotent precursors, primordial germ cells (PGCs) before and after epigenetic reprogramming, and spermatogonia/spermatogonial stem cells (SSCs). Although epigenetic reprogramming, including genome-wide DNA de-methylation, creates broadly open chromatin with abundant enhancer-like signatures, the augmented chromatin insulation safeguards transcriptional fidelity. These insulatory constraints are then erased en masse for spermatogonial development. Notably, despite distinguishing epigenetic programming, including global DNA re-methylation, the PGCs-to-spermatogonia/SThe EMBO journalNucleome programming is required for the foundation of totipotency in mammalian germline development.PMC9251848RGP0057/2018R01 NS111997JP20H05387P01 CA196539CA196539JP18H0261322H04920JPMJER11042018‐193685NS11199717H06098R01 AI118891Yokobayashi SMirny LAYabuta YShimizu SIshikura YNosaka YGarcia BAYamamura ACoradin MKawahira NMurakawa YTachibana KHiraoka YPeters JMOkamoto IMeehan KStocsits RImoto YYamamoto THu BSaitou MOhta HMizuta KIkeda HWutz GKojima YKasahara TMajewski JNagano MUmemura FfalseNucleome programming is required for the foundation of totipotency in mammalian germline development.Germ cells are unique in engendering totipotency, yet the mechanisms underlying this capacity remain elusive. Here, we perform comprehensive and in-depth nucleome analysis of mouse germ-cell development in vitro, encompassing pluripotent precursors, primordial germ cells (PGCs) before and after epigenetic reprogramming, and spermatogonia/spermatogonial stem cells (SSCs). Although epigenetic reprogramming, including genome-wide DNA de-methylation, creates broadly open chromatin with abundant enhancer-like signatures, the augmented chromatin insulation safeguards transcriptional fidelity. These insulatory constraints are then erased en masse for spermatogonial development. Notably, despite distinguishing epigenetic programming, including global DNA re-methylation, the PGCs-to-spermatogonia/S2022-01-01T00:00:00Z2022 Jul2026-05-27T22:42:38.595Z2024-12-03T19:33:51.579ZS-EPMC92518483570312110.15252/embj.2022110600