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In Vivo Monitoring of Ca2+ Uptake into Subcellular Compartments of Mouse Skeletal Muscle

Rüdiger Rudolf1,2,3, Sofie Trajanovska4, David Grant Allen4

  • 1Institute of Molecular and Cell Biology, Mannheim University of Applied Sciences, Mannheim, Germany. r.rudolf@hs-mannheim.de.

Methods in Molecular Biology (Clifton, N.J.)
|January 24, 2019
PubMed

Insights

New methods visualize calcium (Ca2+) signaling in live skeletal muscle during contraction and relaxation. This allows detailed study of muscle function and Ca2+ dynamics at high resolution.

Area of Science:

  • Muscle Physiology
  • Cellular Biology
  • Biophysics

Background:

  • Calcium ions (Ca2+) are critical regulators of skeletal muscle functions, including excitation-contraction coupling and gene expression.
  • Observing Ca2+ signaling in live skeletal muscle is challenging due to rapid Ca2+ transients and muscle contractility.

Purpose of the Study:

  • To describe methods for visualizing Ca2+ signals in skeletal muscle at high spatiotemporal resolution.
  • To enable simultaneous measurement of muscle force alongside Ca2+ dynamics.

Main Methods:

  • Utilizing genetically encoded ratiometric Ca2+ sensors introduced via transfection.
  • Employing two-photon microscopy for high-resolution imaging.
  • Integrating force transducers for simultaneous muscle force measurements.

Main Results:

  • Protocols allow visualization of Ca2+ signals in different subcellular compartments during muscle relaxation-contraction cycles.
  • High spatiotemporal resolution imaging of Ca2+ dynamics is achieved.
  • Simultaneous measurement of muscle force provides functional context to Ca2+ signaling.

Conclusions:

  • The described methods overcome previous limitations in observing Ca2+ signaling in live skeletal muscle.
  • These techniques facilitate detailed investigation of Ca2+ regulation in muscle physiology.
  • Further information is provided on sensor localization and data calibration for semi-quantitative analysis.
Keywords:
CameleonFRETForce transducerMitochondriaMyoplasmRatiometric measurementSarcoplasmic reticulumTwo-photon microscopy

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