Changes in gene expression of caveolin-1 after inflammatory pain using quantitative real-time PCR

This chapter will take the reader through the steps involved in obtaining a gene expression profile using real-time PCR. Real-time PCR is an end-point measure of changes in gene expression that have occurred after a physiological event. It specifically describes the process by which real-time PCR was used to measure caveolin-1 gene expression after an inflammatory pain insult. The method outlines the initial stage of total RNA extraction from the starting material through the main steps involved in producing a gene expression profile including primer design, RNA quantification and cDNA production by reverse transcription and of course the real-time PCR. The caveolin-1 gene measured here is the main component of specialised domains found in the plasma membrane with a primary function described as signal transduction 'hot spots'. The result of this experiment shows that there was a twofold increase in caveolin-1 gene expression in rat dorsal root ganglions 24 h after Complete Freund's Adjuvant administration and a decrease at 48 h.