Methods for Assessing Nuclear Rotation and Nuclear Positioning in Developing Skeletal Muscle Cells

  • Meredith H. Wilson
  • Matthew G. Bray
  • Erika L. F. Holzbaur
Protocol
First Online:
Part of the Methods in Molecular Biology book series (MIMB, volume 1411)

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.

Key words

Skeletal muscle Nuclear positioning Nuclear rotation Angular velocity Rigid body kinematics 
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Notes

Acknowledgments

This work was supported by the National Institutes of Health (P01 GM087253 to E.L.F.H., T32 GM-07229, and T32 AR-053461 to M.H.W.) and the American Heart Association (#13PRE16090007 to M.H.W.).

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Meredith H. Wilson
    • 1
  • Matthew G. Bray
    • 2
  • Erika L. F. Holzbaur
    • 1
  1. 1.Department of Physiology, The Pennsylvania Muscle InstitutePerelman School of Medicine at the University of PennsylvaniaPhiladelphiaUSA
  2. 2.Space Exploration SectorJohns Hopkins Applied Physics LaboratoryLaurelUSA

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