Abstract
This chapter discusses the techniques for observing calcium channels from skeletal muscle sarcoplasmic reticulum in planar lipid bilayers. The sarcoplasmic reticulum (SR) is a specialized intracellular membrane responsible for the distribution of cytoplasmic Ca 2+ . The SR membrane is freely permeable to many small monovalent ions; however, it possesses a stringently regulated system for the release and subsequent reuptake of Ca 2+ . Ca 2+ release from the SR occurs through a calcium channel, which is regulated by external Ca 2+ , Mg 2+ , and adenine nucleotides. The Ca 2+ release channel from SR is a large-conductance divalent-cation-selective channel, which can be incorporated into planar lipid bilayers from heavy terminal cisternae-derived SR fractions. The planar bilayer is a powerful tool for studying ionic currents mediated by single-ion-channel proteins. If conditions are carefully selected, much can be learned about the biophysical and pharmacological properties of individual ion-conducting proteins without the need for rigorous biochemical purification. The lipid bilayer becomes unstable in the presence of large current transients, which are subject to occur during the perfusion procedure. This problem is most noticeable during perfusion of the trans chamber. Any foreign object inserted into the trans chamber (i.e., perfusion tubing) serves as a conduit for these transients.
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