Akiyama Kosuke, Miyashita Takenori, Matsubara Ai, Mori Terushige, Inamoto Ryuhei, Nishiyama Akira, Mori Nozomu
Department of Otolaryngology, Faculty of Medicine, Kagawa University, Kita-gun Miki-cho Ikenobe 1750-1, Kita-gun, Kagawa-ken 761-0793, Japan.
Biochem Biophys Res Commun. 2008 Nov 21;376(3):611-4. doi: 10.1016/j.bbrc.2008.09.052. Epub 2008 Sep 20.
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.
内淋巴囊（ES）是位于颞骨内的一个器官。其解剖位置使得在不受到任何污染的情况下采集ES组织非常困难，有时甚至会因此妨碍对ES进行准确的分子分析。在本研究中，尝试使用激光捕获显微切割技术建立一种新的选择性ES上皮组织采集方法，以获得无污染的纯ES RNA。通过针对骨钙素、钙调蛋白H1和NKCC2的三对特异性引物对进行逆转录聚合酶链反应（RT-PCR），验证了该方法的有效性，这三种蛋白分别是骨、平滑肌和肾/ES细胞中的特异性蛋白。从RT-PCR结果可以看出，新方法具有高特异性和足够的灵敏度。新方法被认为是无污染采集ES的最佳方法，它将有助于未来对ES的分析。
