Biological Procedures Online

, Volume 10, Issue 1, pp 58–65

Cell culture-based analysis of postsynaptic membrane assembly in muscle cells

Open Access
Article

DOI: 10.1251/bpo143

Cite this article as:
Teressa, G. & Prives, J. Biol. Proced. Online (2008) 10: 58. doi:10.1251/bpo143

Abstract

We report a method for studying postsynaptic membrane assembly utilizing the replating of aneural cultures of differentiated skeletal muscle cells onto laminin-coated surfaces. A significant limitation to the current cell culturebased approaches has been their inability to recapitulate the multistage surface acetylcholine receptor (AChR) redistribution events that produce complex AChR clusters found at the intact neuromuscular junction (NMJ). By taking advantage of the ability of substrate laminin to induce advanced maturation of AChR aggregates on the surface of myotubes, we have developed a secondary-plating method that allows more precise analysis of the signaling events connecting substrate laminin stimulation to complex AChR cluster formation. We validate the utility of this method for biochemical and microscopy studies by demonstrating the roles of RhoGTPases in substrate laminin-induced complex cluster assembly.

Indexing terms

Acetylcholine receptors Muscle Agrin Laminin Synapse 

Abbreviations

AChR

acetylcholine receptor

EGFP

enhanced green fluorescent protein

NMJ

neuromuscular junction

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Copyright information

© Springer 2008

Authors and Affiliations

  1. 1.Department of Pharmacological SciencesStony Brook UniversityStony BrookUSA
  2. 2.Medical Scientist Training ProgramStony Brook UniversityStony BrookUSA

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