一种新型单个神经细胞的RT-PCR技术研究

A novel technique used in the RT-PCR of single neurons

提出一种新型可重复实现单细胞的RT-PCR的实验技术,扩增目的基因.应用玻璃微电极提取单个神经细胞,裂解获得mRNA,利用RT-PCR技术检测单个神经细胞的离子通道等基因的表达.应用此技术可以灵敏地检测特定电生理现象的分子生物学基础.此法可以比较大鼠背根神经节中大细胞内HCN1的mRNA表达量.实验结果表明:扩增产物的量足够用于后续实验,而且3个重复的均一性也很好,充分表明了该研究提出的实验技术的可行性.

It is very difficult to complete the application of RT-PCR based on a single cell because of the low level of mRNA.This study is to explore a novel experimental technique to achieve a repeatable process of the single cell RT-PCR,which could be used to characterize the biophysical properties of ion channels genes in a single neuron. The single cell was picked by a glass micropipette,followed by a standard RT-PCR to explore the expression levels of ion channel genes.Single-cell RT-PCR could serve as a precise detector to analyze the molecular basis of the special electrophysiological characteristics. Using the method proposed in the article can be repeated to obtain the expression of HCN1 and HPRT in the big cell of rat dorsal root ganglion. It is enough to continues the next experiment,which proves the feasibility of this technique.