PCR AND PATCH-CLAMP ANALYSIS OF SINGLE NEURONS

The combination of patch-clamp and molecular biology techniques has ma de it possible to characterize the pharmacological and biophysical pro perties of ion channels in single neurons and to screen for expression of specific mRNAs in the same cell. Following whole-cell recording, t he cytoplasm of the cell is harvested, and RNA is reverse transcribed into cDNA and amplified in PCR with primers specific for individual io n channel subunits. Additional experiments can then be designed to rel ate structure and function at the protein level more directly, since c ells appear to regulate the composition of ion channels at least partl y at the posttranscriptional stage.