Study of Caveolae-Dependent Mechanoprotection in Human Muscle Cells Using Micropatterning and Live-Cell Microscopy

Methods Mol Biol. 2020:2169:189-196. doi: 10.1007/978-1-0716-0732-9_17.

Abstract

Caveolae are plasma membrane organelles that are, among many other features, involved in mechanosensing and mechanoprotection. Different tools have been developed to study caveolae-dependent mechanoprotection and had to be adapted to the tissue or cells studied, as these structures are found in almost every type of cells. This chapter focuses on a protocol combining the use of live-cell imaging, micropatterning, hypo-osmotic shock as a mechanical stress, and dyes such as calcein-AM and propidium iodide. We used this protocol for the in vitro study of the effect of mechanical stress on membrane integrity in human muscle cells from patients bearing caveolin-3 mutations.

Keywords: Caveolae; Live-cell imaging; Mechanoprotection; Micropatterning; Muscle cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biomechanical Phenomena / physiology
  • Caveolae / metabolism*
  • Caveolin 3 / genetics
  • Caveolin 3 / metabolism*
  • Cell Line
  • Cell Membrane / metabolism*
  • Fluoresceins / chemistry
  • Humans
  • Image Processing, Computer-Assisted / methods*
  • Microscopy, Video / instrumentation
  • Microscopy, Video / methods*
  • Muscle Cells / metabolism*
  • Muscle Fibers, Skeletal / cytology
  • Muscle Fibers, Skeletal / metabolism*
  • Mutation
  • Osmotic Pressure
  • Propidium / chemistry
  • Stress, Mechanical

Substances

  • CAV3 protein, human
  • Caveolin 3
  • Fluoresceins
  • calcein AM
  • Propidium