PKCγ在中枢神经中的分布与作用机理的探索

发布时间：2018-03-23 01:15

  本文选题：PKCγ　切入点：免疫组织化学染色　出处：《青岛大学》2008年硕士论文　论文类型：学位论文

【摘要】： 蛋白激酶C(protein kinase C,PKC)是重要的细胞内信号转导分子,但PKC的γ亚单位在大鼠神经系统的作用机理目前还不十分明了,对PKC的γ亚单位研究将有助于为阐明PKCγ在痛信息传递和/或调节中的作用机制提供理论依据。本课题从不同角度研究PKCγ的作用机理,以期为阐明PKCγ在痛信息传递和/或调节中的作用机制提供理论依据。内容包括以下四个方面: 第一部分:应用PKCγ特异性抗体的免疫细胞化学染色技术观察蛋白激酶Cγ亚单位(PKCγ)在大鼠神经系统内的定位分布。研究发现PKCγ阳性神经元广泛分布于大鼠神经系统,密集、浓染的PKCγ阳性神经元主要见于大脑皮质、海马、杏仁复合体、小脑皮质、耳蜗腹侧核和耳蜗背侧核。本研究的结果为探讨PKCγ在神经系统信号转导过程中的作用提供了形态学证据。 第二部分:研究大鼠延髓背角和脊髓背角浅层内蛋白激酶Cγ亚单位(PKCγ)阳性神经元向中缝苍白核(NRP)的投射。采用荧光金(FG)逆行追踪和PKCγ免疫荧光组织化学染色相结合的双标记方法。在延髓和脊髓背角的Ⅰ～Ⅲ层内可见FG逆标神经元;PKCγ阳性神经元主要分布于延髓和脊髓背角的Ⅱ层内侧部及Ⅱ、Ⅲ层交界处,Ⅰ、Ⅲ层内较少。提示延髓和脊髓背角浅层向NRP投射的神经元是PKCγ神经元,提示PKCγ在向NRP传递的信息的通路中可能发挥重要作用。 第三部分:应用免疫荧光组织化学双标记染色方法研究PKCγ在延髓和脊髓背角浅层与PV,CB,CR的共存关系。结果提示大部分PKCγ,CB,CR和PV样免疫阳性神经元分布于Ⅱ层,其中也有少量神经元分布于Ⅰ层和Ⅲ层。结论提示PKCγ与钙调蛋白的共存神经元在口面部传递痛信息的通路中可能发挥重要作用。 第四部分:探讨大鼠坐骨神经切断模型脊髓背角内PKCγ的变化及意义。在坐骨神经切断后2、5、10、15、20、30、40、60d,取脊髓背角,应用免疫组织化学方法进行染色,图像分析软件测量并比较脊髓背角手术侧和对照侧的免疫强度。从第2天开始,手术侧脊髓背角L4-L5节断PKCγ免疫阳性反应较对照侧明显增强,差异有显著性(t=3.12,p＜0.05);在第15天时免疫阳性反应最强,是对照组的1.89倍(t=4.85,p＜0.01),然后逐渐减弱,2个月后恢复到正常水平(t=0.91,p＞0.05)。结论提示PKCγ在由神经损伤引起的脊髓背角神经元兴奋性改变中发挥重要作用。
[Abstract]:Protein kinase C(protein kinase is an important intracellular signal transduction molecule, but the mechanism of PKC 纬 subunit in rat nervous system is not well understood. The study of 纬 subunit of PKC will be helpful to clarify the mechanism of PKC 纬 in pain information transmission and / or regulation. In order to provide theoretical basis for elucidating the mechanism of PKC 纬 in pain information transmission and / or regulation, the contents include the following four aspects:. Part one: the localization and distribution of protein kinase C 纬 subunit (PKC 纬) in the nervous system of rats were observed by immunocytochemical staining with specific antibody of PKC 纬. It was found that PKC 纬 positive neurons were widely distributed in the nervous system of rats. Highly stained PKC 纬 positive neurons were found mainly in the cerebral cortex, hippocampus, amygdala complex, cerebellar cortex, The results of this study provide morphological evidence for the study of the role of PKC 纬 in the signal transduction of the nervous system. Part 2: the projection of protein kinase C 纬 subunit (PKC 纬) positive neurons in the dorsal medullary horn and the dorsal horn of spinal cord to the nucleus raphe pallidus (NRP) was studied. The retrograde tracing and PKC 纬 immunofluorescence histochemical staining were used to study the expression of protein kinase C 纬 positive neurons in the dorsal horn of medulla oblongata and the superficial layer of dorsal horn of spinal cord. In the lamina 鈪，

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