In my laboratory, two basic experimental approaches are used. The first utilizes primary cultures of isolated SCG neurons for electrophysiological, molecular biological, immunofluorescence and RT-PCR experiments. The main technique used in this system is to examine the well-characterized ion channel modulatory pathways using patch-clamp electro-physiology as an assay for G protein coupled receptor activity. The neurons can be made to express various heterologous proteins (including mGluRs) via intranuclear injection of cDNA. I have recently used this system to examine mGluR-G protein coupling specificity, mGluR desensitization and the regulation of mGluR signaling and trafficking by various associated proteins.
