Target Gene Localization Using Laser-Capture Microdissection and Real-Time Reverse Transcription-Polymerase Chain Reaction Analysis

• Published in: Journal of histotechnology Vol. 28; no. 3; pp. 177 - 180
• Main Authors: Dalmas, Deidre A., Scicchitano, Marshall S., Zimmerman, Dawn, Tierney, Lauren A., Boyce, Rogely W.
• Format: Journal Article
• Language: English
• Published: Taylor & Francis 01.09.2005
• Subjects: in situ hybridization; laser capture microdissection; real-time RT-PCR
• ISSN: 0147-8885; 2046-0236
• DOI: 10.1179/his.2005.28.3.177

Laser-capture microdissection (LCM) followed by realrime reverse transcription polymerase chain reaction (TaqMan®) is an alternative approach for localizing gene targets in tissue sections when in situ hybridization (ISH) is problematic. LCMITaqMan® and nonisotopic ISH were used to qualitatively and quantitatively evaluate mRNA expression of neuromedin U receptor and Rho kinase in sections of human atherosclerotic plaque. LCM/TaqMan® successfully localized Rho kinase to specific tissue elements when ISH was unsuccessful. LCM/TaqMan® is a powerful alternative/adjunct to ISH because it provides quantitative expression data, has greater sensitivity for detecting low abundance transcripts, and is less sensitive to partial RNA degradation. (The J Histotechnol 28:177, 2005) Submitted November 22, 2004; accepted with revisions June 3, 2005