基因芯片联合激光捕获显微切割技术在脊髓背根神经节基因表达研究中的应用

目的探索联合激光捕获显微切割(laser capture microdissection,LCM)与基因芯片技术在大鼠脊髓背根神经节的感觉神经元基因表达研究中的可行性。方法取大鼠L4、L5、L6节段脊髓背根神经节,用LCM技术获取神经细胞,提取总RNA,甲醛变性凝胶电泳鉴定RNA质量,检测纯度和产量,并以实时RT-PCR方法进行验证。结果总RNA的完整性较好,经单轮T7线性扩增后得到足量的aRNA,可用于实时RT-PCR研究和进一步基因芯片杂交实验。结论结合有效的RNA保护方法和线性扩增,LCM可以分离出纯净的背根神经节的感觉神经元,满足下游基因芯片研究的要求。

Combining Laser Capture Microdissection and Microarray to Analyze the Gene Expression of Rat Dorsal Root Ganglion Sensory Neurons

Objective To test the feasibility of applying laser capture microdissection(LCM) combined with microarray technique in studies on expression of sensory neuron genes in dorsal root ganglion(DRG) of rats. Methods Sensory neurons were obtained from frozen sections of rat L₄-L₆ DRGs by using LCM. The quality and yield of total RNA extracted from captured cells were tested in order to determine the impact of frozen-section cutting, staining and capture procedure. Part of the array results was verified by real-time RT-PCR. Results No degradation was detected, and via a single cycle of T7 linear amplification, the yield was sufficient for RT-PCR and microarray analysis. Conclusion Combined with effective RNA protection and linear amplification, LCM can be applied for gene transcriptional analysis of DRG sensory neurons.