A PROCEDURE FOR RT-PCR AMPLIFICATION OF MESSENGER-RNAS ON HISTOLOGICAL SPECIMENS

Detection in combination with localization of low copy gene expression can be difficult to achieve. The use of reverse transcription PCR on tissue sections with a fluorescent marker provides localization of mRN A expression on a cellular level, and when combined with confocal micr oscopy and image analysis it also allows for an estimate of the relati ve intensity of fluorescence. A good example of the application of thi s method is the localization of nerve growth factor (NGF) mRNA express ion in the inner ear NGF through indirect tests appears to be present in this system, yet NGF MRNA could not be localized with in situ hybri dization using radiolabeled riboprobes. Using fluorescent in situ RT-P CR, we can easily detect the presence of NGF MRNA and localize it to s pecific cell types within the maturing inner ear.