title: Localization of sarcomeric proteins during myofibril assembly in cultured mouse primary skeletal myotubes.
abstract: It is important to understand how muscle forms normally in order to understand muscle diseases that result in abnormal muscle formation. Although the structure of myofibrils is well understood, the process through which the myofibril components form organized contractile units is not clear. Based on the staining of muscle proteins in avian embryonic cardiomyocytes, we previously proposed that myofibrils formation occurred in steps that began with premyofibrils followed by nascent myofibrils and ending with mature myofibrils. The purpose of this study was to determine whether the premyofibril model of myofibrillogenesis developed from studies developed from studies in avian cardiomyocytes was supported by our current studies of myofibril assembly in mouse skeletal muscle. Emphasis was on establishing how the key sarcomeric proteins, F-actin, nonmuscle myosin II, muscle myosin II, and α-actinin were organized in the three stages of myofibril assembly. The results also test previous reports that nonmuscle myosins II A and B are components of the Z-bands of mature myofibrils, data that are inconsistent with the premyofibril model. We have also determined that in mouse muscle cells, telethonin is a late assembling protein that is present only in the Z-bands of mature myofibrils. This result of using specific telethonin antibodies supports the approach of using YFP-tagged proteins to determine where and when these YFP-sarcomeric fusion proteins are localized. The data presented in this study on cultures of primary mouse skeletal myocytes are consistent with the premyofibril model of myofibrillogenesis previously proposed for both avian cardiac and skeletal muscle cells.
authors.affiliation: Department of Medicine, SUNY Upstate Medical University, Syracuse, New York.
authors.forename: Jennifer
authors.initials: J
authors.lastname: White
authors.forename: Marietta V
authors.initials: MV
authors.lastname: Barro
authors.forename: Helen P
authors.initials: HP
authors.lastname: Makarenkova
authors.forename: Joseph W
authors.initials: JW
authors.lastname: Sanger
authors.forename: Jean M
authors.initials: JM
authors.lastname: Sanger
mesh_terms: D000185:Actinin / Q000378:metabolism; D000199:Actins / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D064211:Connectin / Q000378:metabolism; D051379:Mice; D024510:Muscle Development; D018485:Muscle Fibers, Skeletal / Q000378:metabolism / Q000502:physiology*; D009124:Muscle Proteins / Q000378:metabolism*; D009210:Myofibrils / Q000378:metabolism / Q000502:physiology*; D024507:Nonmuscle Myosin Type IIA / Q000378:metabolism; D024501:Nonmuscle Myosin Type IIB / Q000378:metabolism; D061251:Primary Cell Culture; D012518:Sarcomeres / Q000378:metabolism / Q000502:physiology*
chemical_list: D000199:Actins; D064211:Connectin; D009124:Muscle Proteins; C577294:Tcap protein, mouse; D000185:Actinin; D024507:Nonmuscle Myosin Type IIA; D024501:Nonmuscle Myosin Type IIB
keywords: myofibrillogenesis; nonmuscle myosin II; premyofibrils; telethonin; α-actinin