Microtransplantation of acetylcholine receptors from normal or denervated rat skeletal muscles to frog oocytes

J Physiol. 2011 Mar 1;589(Pt 5):1133-42. doi: 10.1113/jphysiol.2010.202994. Epub 2011 Jan 4.

Abstract

Cell membranes, carrying neurotransmitter receptors and ion channels, can be 'microtransplanted' into frog oocytes. This technique allows a direct functional characterization of the original membrane proteins, together with any associated molecules they may have, still embedded in their natural lipid environment. This approach has been previously demonstrated to be very useful to study neurotransmitter receptors and ion channels contained in cell membranes isolated from human brains. Here, we examined the possibility of using the microtransplantation method to study acetylcholine receptors from normal and denervated rat skeletal muscles. We found that the muscle membranes, carrying their fetal or adult acetylcholine receptor isoforms, could be efficiently microtransplanted to the oocyte membrane, making the oocytes become sensitive to acetylcholine. These results show that oocytes injected with skeletal muscle membranes efficiently incorporate functional acetylcholine receptors, thus making the microtransplantation approach a valuable tool to further investigate receptors and ion channels of human muscle diseases.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylcholine / pharmacology
  • Animals
  • Calcium Channels / metabolism
  • Cell Membrane / drug effects
  • Cell Membrane / metabolism*
  • Dose-Response Relationship, Drug
  • Electrophysiology
  • Muscle, Skeletal / drug effects
  • Muscle, Skeletal / innervation
  • Muscle, Skeletal / metabolism*
  • Oocytes / drug effects
  • Oocytes / metabolism
  • Rats
  • Rats, Wistar
  • Receptors, Nicotinic / metabolism*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Xenopus laevis

Substances

  • Calcium Channels
  • Receptors, Nicotinic
  • Acetylcholine