摘要
目的 发展基于颞骨火棉胶切片的突变基因定位方法。方法 选取5例PCR证实具有线粒体DNA(mtDNA)4977缺失的老年性聋颞骨切片,5例对照颞骨切片无mtDNA4977缺失。应用作者首先发展的颞骨火棉胶切片原位杂交技术进行mtDNA4977缺失的定位研究。结果 5例PCR证实具有mtDNA4977缺失的老年性聋颞骨切片中,3例经原位杂交获得代表mtDNA4977缺失的阳性信号,分布于耳蜗、内听道的神经细胞及神经纤维中。5例对照颞骨中未见阳性杂交信号。结论 耳蜗是由多种不同形态、功能的细胞组成,火棉胶切片的原位杂交技术可以定位特殊基因或基因变化于耳蜗内特定的细胞群,为在分子细胞水平研究聋病的发病机制提供了有力的工具。
Purpose To develop localization method of mutated genes in cellodin embedded temporal bone sections. Methods Five cases of temporal bone with mtDNA4977 deletion and five cases without mtDNA4977 deletion identified by nested PCR were chosen in this study and a new technique of in situ hybridization in celloiding embedded temporal bone sections firstly developed by author was used to localize the mtDNA4977 deletion. Results The positive signal representing mtDNA4977 deletion was found in 3 of 5 cases of temporal bone sections with mtDNA4977 deletion proven by PCR and in none of 5 cases without mtDNA4977 deletion. Conclusion Cochlea is constituted by a great number of variable shaped and functional cells. The technique of in situ hybridization in celloidin embedded temporal bone section can detect and distinguish the specific gene or mutated gene among the different cells in cochlea. It provides a powerful tool to study the etiologic mechanism of deafness at the molecular cellular level.
出处
《中华耳科学杂志》
CSCD
2003年第2期22-25,共4页
Chinese Journal of Otology
关键词
颞骨
火棉胶
切片
原位杂交
Temporal bone, Celloidin, Section, In situ Hybridization
