Nestin阳性或阴性胆碱能神经元TrkA受体表达差异的研究

发布时间：2018-05-10 09:03

本文选题：基底前脑 + 巢蛋白　；参考：《大理大学》2017年硕士论文

【摘要】：目的研究健康成年SD大鼠基底前脑巢蛋白(Nestin)阳性或阴性胆碱能神经元神经生长因子受体表达差异,探索两种神经元功能差异的可能机制。方法选取体质量在200~250g,经正常饲养的成年大鼠40只,不限性别,使用多聚甲醛灌注固定后用大鼠脑切片模具取脑,石蜡切片分为单标组和双标组,以及对照组。单标组是:Nestin、ChAT、TrkA,双标组是Nestin和TrkA;ChAT和TrkA组。对荧光双标组用激光共聚焦显微镜观测三种神经元在基底前脑共表达情况。使用免疫组织化学方法、显微镜检测Nestin、ChAT、TrkA于基底前脑内侧隔核、斜角带核垂直支、斜角带核水平支分布以及阳性反应物的表达数量。统计Nestin、TrkA、ChAT三种胆碱能神经元在基底前脑相同区域(MS、vDB、hDB)的细胞数目、ChAT免疫阳性神经元表达TrkA数目、Nestin阳性神经元表达TrkA数目,推测出Nestin阴性神经元表达TrkA数目,使用统计学均数±标准差(`X±S)进行分析。结果实验结果表明,在大鼠基底前脑MS-DBB区域有明显TrkA阳性细胞,TrkA、Nestin胆碱能神经元以及ChAT神经元有相同区域共定位表现,三种神经元相间分布在大鼠基底前脑MS-DBB。数据显示:ChAT免疫阳性神经元数量高于Nestin阳性神经元数量,Nestin几乎完全双标ChAT;有大约90%以上的Nestin阳性神经元表达TrkA,ChAT阳性神经元表达TrkA的量约占70%左右,Nestin阴性神经元也表达TrkA。结论通过荧光双标染色和激光共聚焦显微镜观察得出,Nestin两种神经元能表达TrkA,但是巢蛋白的两种神经元表达TrkA数量是不同的,Nestin阳性神经元表达TrkA的数量高于Nestin阴性神经元表达TrkA的数量。证实巢蛋白的两种神经元表达神经生长因子受体上存在差异,提示巢蛋白阳性神经元能与更多的神经生长因子结合,Nestin阳性或Nestin阴性两种胆碱能神经元在可塑性、兴奋性、抗损伤功能等方面有差异,其机制可能是因为神经生长因子受体表达量的不同而引起的。
[Abstract]:Objective to study the difference of nerve growth factor receptor (NGF) expression in cholinergic neurons of healthy adult SD rats with or without nestin (Nestin) and to explore the possible mechanism of the difference between the two neuronal functions. Methods Forty adult rats with a body weight of 200 ~ 250g were selected and were randomly divided into two groups: single labeled group, double labeled group and control group. The rats were fixed with paraformaldehyde and fixed with the molds of rat brain slice after infusing, and the paraffin sections were divided into single label group and double labeled group, as well as the control group. The single group was Nestin and TrkAc Chat TrkA, and the double labeled group was Nestin and TrkAn chat and TrkA group. The co-expression of three kinds of neurons in basal forebrain was observed by laser confocal microscope in double labeled group. Immunohistochemical method was used to detect the distribution of TrkA in the medial septal nucleus of the basal forebrain, the vertical branch of the diagonal band nucleus, the horizontal branch of the diagonal band nucleus and the expression of positive reactants. The number of the three kinds of cholinergic neurons in the same area of the basal forebrain, the number of chat immunoreactive neurons expressing TrkA and the number of nestin positive neurons expressing TrkA were counted, and the number of Nestin negative neurons expressing TrkA was inferred. Statistical mean 卤standard deviation (`X 卤S) was used to analyze. Results the results showed that there were significant TrkA positive cholinergic neurons and ChAT neurons in the MS-DBB region of the basal forebrain of rats. The three neurons were distributed in MS-DBB of the basal forebrain of rats. The data showed that the number of Nestin immunoreactive neurons was higher than that of Nestin positive neurons, and about 90% of the Nestin positive neurons expressed TrkA, and about 70% of the nestin negative neurons also expressed TrkA. Conclusion two kinds of nestin neurons can express TrkA by fluorescence double labeling and laser confocal microscopy, but the number of TrkA expression in nestin positive neurons is higher than that in Nestin negative neurons. The number of neurons expressing TrkA. It was confirmed that there were differences in the expression of nerve growth factor receptors between the two nestin neurons, suggesting that nestin positive neurons and more cholinergic neurons with nestin binding or Nestin negative neurons could exhibit plasticity and excitability. The mechanism may be due to the different expression of nerve growth factor receptor (NGF).
【学位授予单位】：大理大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R338

【参考文献】