本文選題：肌萎縮側(cè)索硬化 + 神經(jīng)前體細(xì)胞　； 參考：《南昌大學(xué)》2014年碩士論文

【摘要】：目的： 采用G93A-SOD1轉(zhuǎn)基因小鼠作為肌萎縮側(cè)索硬化（ALS）動(dòng)物模型，觀察分析正常生理?xiàng)l件下和ALS病理?xiàng)l件下成年小鼠脊髓神經(jīng)前體細(xì)胞（NPCs）的分布、增殖、遷移途徑和分化方向以及HoxB9的表達(dá)情況。 方法： 構(gòu)建B6SJL-Tg（SOD1*G93A）1Gur/NJU轉(zhuǎn)基因小鼠動(dòng)物模型，用PCR技術(shù)檢測(cè)和篩選轉(zhuǎn)基因小鼠動(dòng)物模型。采用熒光免疫組織化學(xué)雙標(biāo)記技術(shù)，分別標(biāo)記NPCs、神經(jīng)元細(xì)胞、星形膠質(zhì)細(xì)胞、少突膠質(zhì)細(xì)胞、HoxB9基因表達(dá)的細(xì)胞。每一解剖區(qū)同時(shí)進(jìn)行雙重?zé)晒饷庖呓M織化學(xué)染色，在顯微鏡下觀察，并用圖像處理技術(shù)將不同染色的陽(yáng)性細(xì)胞兩兩疊加成像，觀察分析陽(yáng)性細(xì)胞分布特征，，計(jì)算各種陽(yáng)性細(xì)胞數(shù)。并統(tǒng)計(jì)分析相關(guān)解剖區(qū)NPCs、神經(jīng)元細(xì)胞和神經(jīng)膠質(zhì)細(xì)胞的數(shù)量及HoxB基因家族表達(dá)細(xì)胞類型和表達(dá)量的變化。 結(jié)果： 1、正常成年小鼠脊髓組織內(nèi)存在NPCs，并主要分布于中央管周圍區(qū)域，而在灰質(zhì)區(qū)域內(nèi)少量散在分布。不同節(jié)段間中央管周圍區(qū)域的NPCs在數(shù)量上存在頸段＞腰段＞胸段的規(guī)律。 2、在ALS病理狀態(tài)下發(fā)病后的小鼠脊髓內(nèi)存在NPCs的大量增殖，且在各個(gè)解剖區(qū)域均呈現(xiàn)出進(jìn)展組＞起病組＞未發(fā)病組的規(guī)律。 3、在ALS病理狀態(tài)下小鼠脊髓灰質(zhì)（灰質(zhì)前角、后角、側(cè)角區(qū)域）內(nèi)增殖的NPCs與Anti-GFAP antibody存在共免疫反應(yīng)性。提示增殖的NPCs主要分化為星形膠質(zhì)細(xì)胞。 4、在正常和ALS病理狀態(tài)下的小鼠脊髓內(nèi)均不存在HoxB9基因表達(dá)，提示在成年小鼠脊髓內(nèi)HoxB9基因已不再表達(dá)，且在ALS病理?xiàng)l件下不能激活其表達(dá)。 結(jié)論： 1、在正常生理?xiàng)l件下成年小鼠脊髓內(nèi)NPCs主要分布于中央管周圍區(qū)域內(nèi)，而在ALS病理狀態(tài)下，可刺激成年小鼠脊髓內(nèi)NPCs隨疾病的進(jìn)展大量增殖并向著損傷區(qū)域遷移，并且灰質(zhì)區(qū)域內(nèi)增殖的NPCs大多分化為星形膠質(zhì)細(xì)胞。 2、HoxB9基因在正常及ALS成年小鼠脊髓內(nèi)均不再表達(dá)。
[Abstract]:Objective:G93A-SOD1 transgenic mice were used as animal models of amyotrophic lateral sclerosis (ALS) to observe and analyze the distribution and proliferation of neural precursor cells of spinal cord of adult mice under normal physiological conditions and ALS pathological conditions.Migration pathway and differentiation direction and expression of HoxB9.Methods:The animal model of B6SJL-Tg(SOD1*G93A)1Gur/NJU transgenic mice was constructed, and the transgenic mice model was detected and screened by PCR technique.NPCs, neuron cells, astrocytes and oligodendrocytes were labeled with double labeling of HoxB9 gene.At the same time, each anatomic area was stained with double fluorescence immunohistochemical staining, observed under microscope, and the positive cells with different staining were superposed by image processing technique to observe and analyze the distribution of positive cells.The number of positive cells was calculated.The number of NPCs, neuronal cells and glial cells in the related anatomical region, and the changes of HoxB gene family expression cell types were analyzed statistically.Results:1. NPCs were found in the spinal cord of normal adult mice and were mainly distributed around the central canal, while a few were scattered in the gray matter.The number of NPCs in the region around the central canal between different segments was the rule of cervical segment > lumbar segment > thoracic segment.(2) there was a large number of proliferation of NPCs in the spinal cord of the mice after the onset of ALS, and the regularity of NPCs proliferation was observed in all anatomical regions: progressive group > onset group > non-diseased group.3. The proliferative NPCs and Anti-GFAP antibody in the gray matter (anterior horn, posterior horn, lateral horn region) of mouse spinal cord were co-immunoreactive in the pathological state of ALS.These results suggest that proliferative NPCs mainly differentiates into astrocytes.4. There was no expression of HoxB9 gene in the spinal cord of normal and ALS mice, suggesting that HoxB9 gene was no longer expressed in the spinal cord of adult mice and could not be activated under the pathological condition of ALS.Conclusion:1. Under normal physiological conditions, NPCs in the spinal cord of adult mice was mainly distributed in the area around the central canal, while in the pathological state of ALS, it could stimulate the proliferation of NPCs in the spinal cord of adult mice with the progression of the disease and migrate to the injured area.And most of the proliferative NPCs in gray matter area differentiated into astrocytes.2 the HoxB9 gene was no longer expressed in the spinal cord of normal and ALS adult mice.
【學(xué)位授予單位】：南昌大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R744.8

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相關(guān)期刊論文 前3條

1 徐仁O5;;肌萎縮側(cè)索硬化的診斷、鑒別診斷[J];國(guó)外醫(yī)學(xué)(老年醫(yī)學(xué)分冊(cè));2006年03期

2 徐仁O5;;肌萎縮側(cè)索硬化的治療研究進(jìn)展[J];國(guó)外醫(yī)學(xué)(老年醫(yī)學(xué)分冊(cè));2006年04期

3 徐仁O5;陶玉慧;吳成斯;楊云珠;易娟;;肌萎縮側(cè)索硬化的病因研究進(jìn)展[J];臨床薈萃;2007年01期

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