The power of single channel recording and analysis: its application to ryanodine receptors in lipid bilayers.

Abstract

1. Since the inception of the patch-clamp technique, single-channel recording has made an enormous impact on our understanding of ion channel function and its role in membrane transport and cell physiology. 2. However, the impact of single-channel recording methods on our understanding of intracellular Ca2+ regulation by internal stores is not as broadly recognized. There are several possible reasons for this. 3. First, ion channels in the membranes of intracellular organelles are not directly accessible to patch pipettes, requiring other methods that are not as widely known as the patch-clamp techniques. 4. Second, bulk assays for channel activity have proved successful in advancing our knowledge of Ca2+ handling by intracellular stores. These assays include Ca2+ imaging, ryanodine binding assays and measurements of muscle tension and Ca2+ release and uptake by vesicles that have been isolated from internal stores. 5. The present review describes methods used for single- channel recording and analysis, as applied to the calcium release channels in striated muscle, and details some of the unique contributions that single-channel recording and analysis have made to our current understanding of the release of Ca2+ from the internal stores of muscle. 6. With this in mind, the review focuses on three aspects of channel function and shows how single-channel investigations have led to an improved understanding of physiological processes in muscle. 7. Finally, the review describes some of the latest improvements in membrane technology that will underpin future advances in single-channel recording.