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Methods for Assessing Nuclear Rotation and Nuclear Positioning in Developing Skeletal Muscle Cells
Meredith H Wilson1, Matthew G Bray2, Erika L F Holzbaur3
1Department 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.
Methods in Molecular Biology (Clifton, N.J.)
|May 6, 2016
Summary
This study details methods for measuring nuclear rotation speed in muscle cells using live-cell imaging. It also provides protocols for analyzing how nuclei are distributed within these cells.
Area of Science:
- Cell Biology
- Biophysics
- Muscle Physiology
Background:
- Skeletal muscle cells are multinucleated syncytia with regularly positioned nuclei.
- Microtubule motor proteins, kinesin-1 and cytoplasmic dynein, drive nuclear distribution during development.
- Nuclear movement involves both translocation and three-dimensional rotation.
Purpose of the Study:
- To describe methods for quantitatively assessing nuclear rotation speed in cultured myotubes.
- To provide protocols for analyzing nuclear distribution within myotubes.
Main Methods:
- Live-cell imaging techniques were employed to observe nuclear dynamics.
- Rigid body kinematic analyses were used to quantify the speed of nuclear rotation.
- Protocols for assessing nuclear distribution were developed.
Main Results:
- The study presents methodologies for measuring nuclear rotation speed.
- Protocols for analyzing nuclear distribution in cultured myotubes are detailed.
Conclusions:
- Quantitative assessment of nuclear rotation and distribution is crucial for understanding muscle cell development and function.
- The described methods facilitate further research into the mechanisms of nuclear positioning in skeletal muscle.
