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


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.
Kimura
Yurika
Y
The Department of Otolaryngology, Graduate School, Tokyo Medical and Dental University, Tokyo, Japan.
Kouda
Hiroko
H
Eishi
Yoshinobu
Y
Kobayashi
Daisuke
D
Suzuki
Yoshimi
Y
Ishige
Ikuo
I
Iino
Yukiko
Y
Kitamura
Ken
K
D004252:DNA Mutational Analysis; D004272:DNA, Mitochondrial / Q000032:analysis / Q000235:genetics / Q000302:isolation & purification*; D007758:Ear, Inner / Q000737:chemistry* / Q000166:cytology; D034381:Hearing Loss / Q000235:genetics; D006801:Humans; D007834:Lasers; D042282:Microdissection / Q000295:instrumentation / Q000379:methods*; D009154:Mutation; D016133:Polymerase Chain Reaction / Q000379:methods; D013701:Temporal Bone / Q000737:chemistry / Q000166:cytology
D004272:DNA, Mitochondrial