A new approach for selective rat endolymphatic sac epithelium collection to obtain pure specific RNA

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• 作者：Kosuke Akiyama ; Takenori Miyashita ; Ai Matsubara ; Terushige Mori ; Ryuhei Inamoto ; Akira Nishiyama ; Nozomu Mori
• 关键词：Endolymphatic sac ; Laser capture microdissection ; Tissue collection ; RNA ; Tissue contamination ; Rat ; Polymerase chain reaction ; Temporal bone ; Inner ear ; Molecular analysis
• 刊名：Biochemical and Biophysical Research Communications
• 出版年：2008
• 出版时间：21 November 2008
• 年：2008
• 卷：376
• 期：3
• 页码：611-614
• 全文大小：690 K

文摘

The endolymphatic sac (ES) is an organ that is located in the temporal bone. Its anatomical location makes ES tissue collection without any contamination very difficult, and sometimes accurate molecular analyses of the ES are prevented due to this matter. In the present study, a new selective ES epithelial tissue collection method was attempted using laser capture microdissection to obtain pure ES RNA without any contamination. The validity of this method was demonstrated by RT-PCR with three specific primer pairs against osteocalcin, calponin H1, and NKCC2, which are specific proteins in bone, smooth muscle, and kidney/ES cells, respectively. From the RT-PCR results, the high specificity and sufficient sensitivity of the new method was indicated. It is considered that the new method is optimal for ES collection without contamination and it will be able to contribute to future analyses of the ES.