Amplification of RNA from archival human temporal bone sections.
BACKGROUND
The amplification of DNA from celloidin-embedded human temporal bone sections by polymerase chain reaction (PCR) has been applied to some auditory diseases, such as herpes zoster oticus and hearing loss caused by the mutations of mitochondrial DNA. However, few studies have reported detection of RNA from temporal bone sections.


OBJECTIVE
Because RNA analysis from temporal bone sections may elucidate the development of the diseases in the auditory, vestibular, and facial nerves, the authors investigated whether RNA in these sections can be amplified by reverse transcription (RT)-PCR.


METHODS
Sections that were formalin-fixed, decalcified, and embedded between 1972 and 1986 were used. Nucleic acid was extracted from the celloidin-embedded temporal bone sections and subjected to RT-PCR. Human alpha-tubulin RNA was reverse transcribed to cDNA and amplified by nested PCR using two sets of primers that were designed to distinguish cDNA from genomic DNA based on the presence of an intron between the primers.


RESULTS
Human alpha-tubulin RNA was detected in 11 of 14 temporal bone sections (79%) by RT-PCR. RNA was detected in even the oldest sections, which were processed in 1972.


CONCLUSIONS
These results indicate that RNA can be analyzed from archival celloidin-embedded human temporal bone sections.
Ohtani
F
F
Department of Otolaryngology, Hokkaido University School of Medicine, Sapporo, Japan.
Furuta
Y
Y
Iino
Y
Y
Inuyama
Y
Y
Fukuda
S
S
D001109:Archives; D017931:DNA Primers / Q000235:genetics; D004272:DNA, Mitochondrial / Q000235:genetics; D004427:Ear Diseases / Q000235:genetics; D006801:Humans; D021141:Nucleic Acid Amplification Techniques; D017354:Point Mutation / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D013701:Temporal Bone / Q000473:pathology*; D014404:Tubulin / Q000235:genetics
D017931:DNA Primers; D004272:DNA, Mitochondrial; D012333:RNA, Messenger; D014404:Tubulin