论文部分内容阅读
目的观察穹隆海马伞切割后不同时间点海马内RUNX1T1 mRNA和蛋白的表达变化及定位。方法行穹隆海马伞切割术,制备模型大鼠。实验分为正常组、切割3d组、切割7d组、切割14d组、切割21d组、切割28d组。1.提取海马组织总mRNA,用Real-time PCR检测大鼠海马组织中RUNX1T1mRNA的表达;2.提取海马组织总蛋白,用Western blotting检测海马内RUNX1T1蛋白的表达;3.行脑冷冻切片,行RUNX1T1免疫荧光检测和Hoechst标记,观察海马齿状回中RUNX1T1/Hoechst双标阳性细胞。结果 Real-time PCR检测显示,切割3d组海马组织中RUNX1T1 mRNA的表达明显上调,其余各组差异不明显;Western blotting检测结果显示,正常组海马中有微量RUNX1T1蛋白表达,而切割3d组海马组织中RUNX1T1蛋白表达量明显上升,并达到高峰,7d时仍有较多的表达,此后,14d、21d、28d组中RUNX1T1蛋白表达很快又恢复到正常水平;免疫荧光检测结果表明,正常组海马齿状回中有少量的RUNX1T1/Hoechst阳性细胞,切割3d组海马齿状回中RUNX1T1/Hoechst阳性细胞数量最多,切割7d组逐渐减少,切割14d组、21d组和28d组与正常组差别不明显。结论穹隆海马伞切割后海马内RUNX1T1mRNA和蛋白表达在早期呈短暂性上调,并定位于海马齿状回颗粒下层,推测可能与海马神经再生过程中神经干细胞向神经元分化有关。
Objective To observe the expression and localization of RUNX1T1 mRNA and protein in the hippocampus of the hippocampus at different time points after the dome fornivated dome was cut. Methods Forpalpatectomy of the hippocampus to prepare model rats. The experiment was divided into normal group, cutting 3d group, cutting 7d group, cutting 14d group, cutting 21d group, cutting 28d group. 1. The total mRNA of hippocampus was extracted and the expression of RUNX1T1 mRNA in hippocampus was detected by Real-time PCR. 2. The total protein in hippocampus was extracted and the expression of RUNX1T1 protein in hippocampus was detected by Western blotting.3. RUNX1T1 immunofluorescence and Hoechst labeling were used to detect RUNX1T1 / Hoechst double positive cells in hippocampal dentate gyrus. Results Real-time PCR showed that the expression of RUNX1T1 mRNA in the hippocampus of the 3d group was significantly up-regulated, while the other groups showed no significant difference. Western blotting showed that there was trace RUNX1T1 protein expression in the normal hippocampus, The expression of RUNX1T1 protein increased significantly and peaked at 7d, and the expression of RUNX1T1 protein rapidly returned to normal after 14d, 21d and 28d. The results of immunofluorescence showed that the expression of RUNX1T1 in hippocampus There was a small amount of RUNX1T1 / Hoechst positive cells in the dentate gyrus. The number of RUNX1T1 / Hoechst positive cells in the dentate gyrus of the 3d group was the highest, and the number of RUNX1T1 / Hoechst positive cells was gradually decreased in the dentate gyrus. The difference was not obvious between the 14th and 21st groups . Conclusions The expression of RUNX1T1mRNA and protein in the hippocampus of the hippocampus after transient fornix transection were transiently up-regulated in the hippocampus, and located in the inferior granular layer of hippocampal dentate gyrus, which may be related to the differentiation of neural stem cells to neurons during hippocampal neurogenesis.