title: Myocyte differentiation generates nuclear invaginations traversed by myofibrils associating with sarcomeric protein mRNAs.
abstract: Certain types of cell both in vivo and in vitro contain invaginated or convoluted nuclei. However, the mechanisms and functional significance of the deformation of the nuclear shape remain enigmatic. Recent studies have suggested that three types of cytoskeleton, microfilaments, microtubules and intermediate filaments, are involved in the formation of nuclear invaginations, depending upon cell type or conditions. Here, we show that undifferentiated mouse C2C12 skeletal muscle myoblasts had smoothsurfaced spherical or ellipsoidal nuclei, whereas prominent nuclear grooves and invaginations were formed in multinucleated myotubes during terminal differentiation. Conversion of mouse fibroblasts to myocytes by the transfection of MyoD also resulted in the formation of nuclear invaginations after differentiation. C2C12 cells prevented from differentiation did not have nuclear invaginations, but biochemically differentiated cells without cell fusion exhibited nuclear invaginations. Thus, biochemical differentiation is sufficient for the nuclear deformation. Although vimentin markedly decreased both in the biochemically and in the terminally differentiated cells, exogenous expression of vimentin in myotubes did not rescue nuclei from the deformation. On the other hand, non-striated premyofibrils consisting of sarcomeric actinmyosin filament bundles and cross-striated myofibrils traversed the grooves and invaginations. Time-lapse microscopy showed that the preformed myofibrillar structures cut horizontally into the nuclei. Prevention of myofibril formation retarded the generation of nuclear invaginations. These results indicate that the myofibrillar structures are, at least in part, responsible for the formation of nuclear grooves and invaginations in these myocytes. mRNA of sarcomeric proteins including myosin heavy chain and alpha-actin were frequently associated with the myofibrillar structures running along the nuclear grooves and invaginations. Consequently, the grooves and invaginations might function in efficient sarcomeric protein mRNA transport from the nucleus along the traversing myofibrillar structures for active myofibril formation.
authors.affiliation: Department of Biology, Faculty of Science, and Graduate School of Science and Technology, Chiba University, Yayoicho, Inageku, Chiba, Chiba 263-8522, Japan.
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Abe
authors.forename: Kazunori
authors.initials: K
authors.lastname: Takano
authors.forename: Akiko
authors.initials: A
authors.lastname: Suzuki
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Shimada
authors.forename: Masaki
authors.initials: M
authors.lastname: Inagaki
authors.forename: Naruki
authors.initials: N
authors.lastname: Sato
authors.forename: Takashi
authors.initials: T
authors.lastname: Obinata
authors.forename: Takeshi
authors.initials: T
authors.lastname: Endo
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D002467:Cell Nucleus / Q000648:ultrastructure*; D015536:Down-Regulation; D017404:In Situ Hybridization, Fluorescence; D051379:Mice; D018613:Microscopy, Confocal; D032342:Muscle Cells / Q000648:ultrastructure*; D009124:Muscle Proteins / Q000235:genetics / Q000378:metabolism*; D018482:Muscle, Skeletal; D017570:MyoD Protein / Q000235:genetics / Q000378:metabolism*; D009210:Myofibrils / Q000378:metabolism* / Q000648:ultrastructure; D009218:Myosins / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D014162:Transfection; D014746:Vimentin / Q000378:metabolism
chemical_list: D000199:Actins; D009124:Muscle Proteins; D017570:MyoD Protein; C057363:MyoD1 myogenic differentiation protein; D012333:RNA, Messenger; D014746:Vimentin; D009218:Myosins