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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。
OBJECTIVE: To develop a method for the localization of mutated genes based on temporal bone polymethylmethacrylate. Methods 5 cases of senile deaf temporal bone slices with mitochondrial DNA 4977 deletion were selected and 5 cases of control temporal bone slices without mtDNA4977 deletion. Application of the first development of temporal bone collodione in situ hybridization in situ hybridization mtDNA4977 deletion localization. Results In 5 cases of senile deaf temporal bone slices with mtDNA4977 deletion confirmed by 5 PCR, positive signals of mtDNA4977 deletion were obtained by in situ hybridization and distributed in the cochlear, auditory nerve cells and nerve fibers. There was no positive hybridization signal in 5 cases of temporal bone. Conclusions Cochlear implants are composed of many different morphological and functional cells. In situ hybridization with collombellum slides can locate specific cell groups in which special genes or genes change in the cochlea, providing a basis for studying the pathogenesis of deafness at the molecular cellular level A powerful tool.