Ohtani F, Furuta Y, Iino Y, Inuyama Y, Fukuda S
Department of Otolaryngology, Hokkaido University School of Medicine, Sapporo, Japan.
Laryngoscope. 1999 Apr;109(4):617-20. doi: 10.1097/00005537-199904000-00018.
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.
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.
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.
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.
These results indicate that RNA can be analyzed from archival celloidin-embedded human temporal bone sections.
通过聚合酶链反应（PCR）从火棉胶包埋的人类颞骨切片中扩增DNA已应用于一些听觉疾病，如耳带状疱疹和线粒体DNA突变引起的听力损失。然而，很少有研究报道从颞骨切片中检测RNA。
由于对颞骨切片进行RNA分析可能有助于阐明听觉、前庭和面神经疾病的发展过程，作者研究了这些切片中的RNA是否可以通过逆转录（RT）-PCR进行扩增。
使用1972年至1986年间福尔马林固定、脱钙并包埋的切片。从火棉胶包埋的颞骨切片中提取核酸，并进行RT-PCR。将人α-微管蛋白RNA逆转录为cDNA，并使用两组引物通过巢式PCR进行扩增，这两组引物设计用于根据引物之间内含子的存在来区分cDNA和基因组DNA。
通过RT-PCR在14个颞骨切片中的11个（79%）中检测到了人α-微管蛋白RNA。即使是1972年处理的最古老的切片中也检测到了RNA。
这些结果表明，可以从存档的火棉胶包埋的人类颞骨切片中分析RNA。
