title: Methods for Assessing Nuclear Rotation and Nuclear Positioning in Developing Skeletal Muscle Cells.
abstract: Skeletal muscle cells are large syncytia, containing hundreds of nuclei positioned regularly along the length of the fiber. During development, nuclei are actively distributed throughout the myotube by the microtubule motor proteins, kinesin-1, and cytoplasmic dynein. Nuclear movement consists of translocation along the long axis of the cell concurrent with three-dimensional rotation of nuclei. In this chapter we describe methods for quantitatively assessing the speed of nuclear rotation in cultured myotubes using live-cell imaging techniques coupled with rigid body kinematic analyses. Additionally, we provide protocols for analyzing nuclear distribution in myotubes.
authors.affiliation: Department of Physiology, The Pennsylvania Muscle Institute, Perelman School of Medicine at the University of Pennsylvania, 638A Clinical Research Building, 415 Curie Boulevard, Philadelphia, PA, 19104-6085, USA.
authors.forename: Meredith H
authors.initials: MH
authors.lastname: Wilson
authors.affiliation: Space Exploration Sector, Johns Hopkins Applied Physics Laboratory, Laurel, MD, 21723, USA.
authors.forename: Matthew G
authors.initials: MG
authors.lastname: Bray
authors.affiliation: Department of Physiology, The Pennsylvania Muscle Institute, Perelman School of Medicine at the University of Pennsylvania, 638A Clinical Research Building, 415 Curie Boulevard, Philadelphia, PA, 19104-6085, USA. holzbaur@mail.med.upenn.edu.
authors.forename: Erika L F
authors.initials: EL
authors.lastname: Holzbaur
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D002467:Cell Nucleus / Q000378:metabolism*; D002478:Cells, Cultured; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry; D051379:Mice; D008856:Microscopy, Fluorescence; D024510:Muscle Development; D018485:Muscle Fibers, Skeletal / Q000166:cytology* / Q000378:metabolism*; D018482:Muscle, Skeletal / Q000166:cytology / Q000196:embryology / Q000378:metabolism; D032446:Myoblasts
keywords: Angular velocity; Nuclear positioning; Nuclear rotation; Rigid body kinematics; Skeletal muscle