title: Specific pattern of cell cycle during limb fetal myogenesis.
abstract: Tight regulation of cell proliferation and differentiation is required to ensure proper growth during development and post-natal life. The source and nature of signals regulating cell proliferation are not well identified in vivo. We investigated the specific pattern of proliferating cells in mouse limbs, using the Fluorescent ubiquitynation-based cell-cycle indicator (Fucci) system, which allowed the visualization of the G1, G1/S transition and S/G2/M phases of the cell cycle in red, yellow or green fluorescent colors, respectively. We also used the retroviral RCAS system to express a Fucci cassette in chick embryos. We performed a comprehensive analysis of the cell cycle state of myogenic cells in fetal limb muscles, adult myoblast primary cultures and isolated muscle fiber cultures using the Fucci transgenic mice. We found that myonuclei of terminally differentiated muscle fibers displayed Fucci red fluorescence during mouse and chick fetal development, in adult isolated muscle fiber (ex vivo) and adult myoblast (in vitro) mouse cultures. This indicated that myonuclei exited from the cell cycle in the G1 phase and are maintained in a blocked G1-like state. We also found that cycling muscle progenitors and myoblasts in G1 phase were not completely covered by the Fucci system. During mouse fetal myogenesis, Pax7+ cells labeled with the Fucci system were observed mostly in S/G2/M phases. Proliferating cells in S/G2/M phases displayed a specific pattern in mouse fetal limbs, delineating individualized muscles. In addition, we observed more Pax7+ cells in S/G2/M phases at muscle tips, compared to the middle of muscles. These results highlight a specific spatial regionalization of cycling cells at the muscle borders and muscle-tendon interface during fetal development.
authors.affiliation: CNRS UMR 7622, IBPS-Developmental Biology Laboratory, F-75005 Paris, France; Sorbonne Universités, UPMC Univ Paris 06, IBPS-Developmental Biology Laboratory, F-75005 Paris, France; Inserm U1156, F-75005 Paris, France.
authors.forename: Joana
authors.initials: J
authors.lastname: Esteves de Lima
authors.affiliation: CNRS UMR 7622, IBPS-Developmental Biology Laboratory, F-75005 Paris, France; Sorbonne Universités, UPMC Univ Paris 06, IBPS-Developmental Biology Laboratory, F-75005 Paris, France; Inserm U1156, F-75005 Paris, France.
authors.forename: Marie-Ange
authors.initials: MA
authors.lastname: Bonnin
authors.affiliation: CNRS UMR 7622, IBPS-Developmental Biology Laboratory, F-75005 Paris, France; Sorbonne Universités, UPMC Univ Paris 06, IBPS-Developmental Biology Laboratory, F-75005 Paris, France; Inserm U1156, F-75005 Paris, France.
authors.forename: Adeline
authors.initials: A
authors.lastname: Bourgeois
authors.affiliation: Inserm U1016, Institut Cochin, Paris, France; CNRS UMR 8104, Paris, France; Université Paris Descartes, Sorbonne Paris Cité, Paris F-75014, France.
authors.forename: Alice
authors.initials: A
authors.lastname: Parisi
authors.affiliation: Inserm U1016, Institut Cochin, Paris, France; CNRS UMR 8104, Paris, France; Université Paris Descartes, Sorbonne Paris Cité, Paris F-75014, France.
authors.forename: Fabien
authors.initials: F
authors.lastname: Le Grand
authors.affiliation: CNRS UMR 7622, IBPS-Developmental Biology Laboratory, F-75005 Paris, France; Sorbonne Universités, UPMC Univ Paris 06, IBPS-Developmental Biology Laboratory, F-75005 Paris, France; Inserm U1156, F-75005 Paris, France. Electronic address: delphine.duprez@upmc.fr.
authors.forename: Delphine
authors.initials: D
authors.lastname: Duprez
mesh_terms: D000818:Animals; D002453:Cell Cycle / Q000502:physiology*; D002467:Cell Nucleus / Q000502:physiology*; D002642:Chick Embryo; D005121:Extremities / Q000196:embryology*; D005333:Fetus / Q000502:physiology*; D007150:Immunohistochemistry; D051379:Mice; D008822:Mice, Transgenic; D008856:Microscopy, Fluorescence / Q000379:methods; D024510:Muscle Development / Q000502:physiology*; D018482:Muscle, Skeletal / Q000502:physiology*; D051763:PAX7 Transcription Factor / Q000378:metabolism; D013234:Stem Cells / Q000378:metabolism; D054875:Ubiquitination
chemical_list: D051763:PAX7 Transcription Factor; C497322:Pax7 protein, mouse
keywords: Cell cycle; Fetal myogenesis; Fucci; MyoD; Pax7