Patch-Clamp and Single-Cell Reverse Transcription–Polymerase Chain Reaction/Microarray Analysis
If gene expression could be analyzed after electrophysiological recording of a single cell, it would enhance our molecular understanding of neuronal physiology and help to interpret the results obtained from patch-clamp recordings. For instance, after completing a recording from a neuron in a brain slice preparation, analysis of the cell’s mRNA could reveal the kind of neurotransmitter the cell used. Detection of an ion channel’s mRNA would provide molecular evidence of the presence of the ion channel as predicted by the electrophysiological experiment. However, the amount of mRNA in single (neuronal) cell is very small. Therefore, analysis of mRNA from a single cell requires special isolation techniques for the mRNA and an accurate amplification step. Recent remarkable advances in molecular biology now allow us to apply reverse transcription–polymerase chain reaction methods to the analysis of specific gene expression as well as microarray analysis of the entire transcriptome from a single cell.
KeywordsEnhance Green Fluorescent Protein Pipette Solution Glass Pipette Orexin Neuron Microarray Gene Expression Analysis