该研究采用激光显微切割技术从小鼠大脑第五皮层捕获少量皮质脊髓投射神经元,并分析其分子生物学特征。首先,提取神经元微量RNA,再借助WT-ovation试剂盒扩增RNA,随后利用合成的cDNA进行荧光定量PCR实验,分析投射神经元核糖体蛋白相关基因的表达情况。结果发现,与大脑皮层其它皮层组织相比,第五皮层投射神经元内部分核糖体蛋白相关基因的表达相对较高。这表明第五皮层投射神经元可能具有活跃的蛋白质合成功能,是核糖体蛋白质的能量场所,借此维持第五皮层脊髓投射神经元长轴突和大胞体的能量需要。
In this study, we captured the small amounts of corticospinal projection neurons (CSPNs) from layer V of the mouse cortical cortex by Laser Capture Microdissection technology, and analyzed its molecular biol- ogy characteristics. RNA was extracted from mouse CSPNs, and then amplified with WT-ovation kit. After cDNA was reverse transcriptted from the amplified RNA, Real-time quantitative PCR was performed. We analyzed the ribosomal protein genes expressions of CSPNs. Our results showed that the mRNA expression levels of some ribosomal proteins within layer V projection neurons were much higher compared with those of other layers. It indicated that the protein synthesis function of layer V projection neurons might be active. It provided a space for energy synthetising of ribosomal proteins. The energy played an essential role for maintaining layer V projection neurons--long axons and big soma.