本文選題：PKCγ　切入點(diǎn)：免疫組織化學(xué)染色　出處：《青島大學(xué)》2008年碩士論文　論文類型：學(xué)位論文

【摘要】： 蛋白激酶C(protein kinase C,PKC)是重要的細(xì)胞內(nèi)信號(hào)轉(zhuǎn)導(dǎo)分子,但PKC的γ亞單位在大鼠神經(jīng)系統(tǒng)的作用機(jī)理目前還不十分明了,對(duì)PKC的γ亞單位研究將有助于為闡明PKCγ在痛信息傳遞和/或調(diào)節(jié)中的作用機(jī)制提供理論依據(jù)。本課題從不同角度研究PKCγ的作用機(jī)理,以期為闡明PKCγ在痛信息傳遞和/或調(diào)節(jié)中的作用機(jī)制提供理論依據(jù)。內(nèi)容包括以下四個(gè)方面: 第一部分:應(yīng)用PKCγ特異性抗體的免疫細(xì)胞化學(xué)染色技術(shù)觀察蛋白激酶Cγ亞單位(PKCγ)在大鼠神經(jīng)系統(tǒng)內(nèi)的定位分布。研究發(fā)現(xiàn)PKCγ陽性神經(jīng)元廣泛分布于大鼠神經(jīng)系統(tǒng),密集、濃染的PKCγ陽性神經(jīng)元主要見于大腦皮質(zhì)、海馬、杏仁復(fù)合體、小腦皮質(zhì)、耳蝸腹側(cè)核和耳蝸背側(cè)核。本研究的結(jié)果為探討PKCγ在神經(jīng)系統(tǒng)信號(hào)轉(zhuǎn)導(dǎo)過程中的作用提供了形態(tài)學(xué)證據(jù)。 第二部分:研究大鼠延髓背角和脊髓背角淺層內(nèi)蛋白激酶Cγ亞單位(PKCγ)陽性神經(jīng)元向中縫蒼白核(NRP)的投射。采用熒光金(FG)逆行追蹤和PKCγ免疫熒光組織化學(xué)染色相結(jié)合的雙標(biāo)記方法。在延髓和脊髓背角的Ⅰ～Ⅲ層內(nèi)可見FG逆標(biāo)神經(jīng)元;PKCγ陽性神經(jīng)元主要分布于延髓和脊髓背角的Ⅱ?qū)觾?nèi)側(cè)部及Ⅱ、Ⅲ層交界處,Ⅰ、Ⅲ層內(nèi)較少。提示延髓和脊髓背角淺層向NRP投射的神經(jīng)元是PKCγ神經(jīng)元,提示PKCγ在向NRP傳遞的信息的通路中可能發(fā)揮重要作用。 第三部分:應(yīng)用免疫熒光組織化學(xué)雙標(biāo)記染色方法研究PKCγ在延髓和脊髓背角淺層與PV,CB,CR的共存關(guān)系。結(jié)果提示大部分PKCγ,CB,CR和PV樣免疫陽性神經(jīng)元分布于Ⅱ?qū)?其中也有少量神經(jīng)元分布于Ⅰ層和Ⅲ層。結(jié)論提示PKCγ與鈣調(diào)蛋白的共存神經(jīng)元在口面部傳遞痛信息的通路中可能發(fā)揮重要作用。 第四部分:探討大鼠坐骨神經(jīng)切斷模型脊髓背角內(nèi)PKCγ的變化及意義。在坐骨神經(jīng)切斷后2、5、10、15、20、30、40、60d,取脊髓背角,應(yīng)用免疫組織化學(xué)方法進(jìn)行染色,圖像分析軟件測量并比較脊髓背角手術(shù)側(cè)和對(duì)照側(cè)的免疫強(qiáng)度。從第2天開始,手術(shù)側(cè)脊髓背角L4-L5節(jié)斷PKCγ免疫陽性反應(yīng)較對(duì)照側(cè)明顯增強(qiáng),差異有顯著性(t=3.12,p＜0.05);在第15天時(shí)免疫陽性反應(yīng)最強(qiáng),是對(duì)照組的1.89倍(t=4.85,p＜0.01),然后逐漸減弱,2個(gè)月后恢復(fù)到正常水平(t=0.91,p＞0.05)。結(jié)論提示PKCγ在由神經(jīng)損傷引起的脊髓背角神經(jīng)元興奮性改變中發(fā)揮重要作用。
[Abstract]:Protein kinase C(protein kinase is an important intracellular signal transduction molecule, but the mechanism of PKC 緯 subunit in rat nervous system is not well understood. The study of 緯 subunit of PKC will be helpful to clarify the mechanism of PKC 緯 in pain information transmission and / or regulation. In order to provide theoretical basis for elucidating the mechanism of PKC 緯 in pain information transmission and / or regulation, the contents include the following four aspects:. Part one: the localization and distribution of protein kinase C 緯 subunit (PKC 緯) in the nervous system of rats were observed by immunocytochemical staining with specific antibody of PKC 緯. It was found that PKC 緯 positive neurons were widely distributed in the nervous system of rats. Highly stained PKC 緯 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 緯 in the signal transduction of the nervous system. Part 2: the projection of protein kinase C 緯 subunit (PKC 緯) 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 緯 immunofluorescence histochemical staining were used to study the expression of protein kinase C 緯 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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