Detection of mitochondrial DNA from human inner ear using real-time polymerase chain reaction and laser microdissection

Abstract

Conclusions. In this study we were able to amplify and analyze extremely small amounts of template DNA from only a few individually dissected cells. We anticipate that this approach will facilitate the detection and analysis of mitochondrial (mt) DNA mutations in specific cell types in the inner ear, which should shed new light on genetic disorders leading to hearing loss. Objective. To isolate mtDNA from selected tissues in the inner ear. Although several methods for extracting DNA from formalin-fixed, celloidin-embedded, archival human temporal bones have been reported, the isolation of DNA from the inner ear by means of laser microdissection has not been previously demonstrated. Material and methods. This was a retrospective study. Temporal bones were obtained from subjects with no known otological history at autopsy. The combined method of laser microdissection and real-time polymerase chain reaction was used to isolate mtDNA from selected tissues in the inner ear. Results. mtDNA could be isolated from the stria vascularis, spiral ligament, spiral ganglion cells and organ of Corti.

• Genetic hearing impairment
• human inner ear
• laser microdissection
• mitochondrial DNA
• real-time polymerase chain reaction