肌萎缩侧索硬化脊髓内源性神经前体细胞的变化及其调控因子HoxB表达改变的研究

发布时间：2018-04-17 05:06

本文选题：肌萎缩侧索硬化 + 神经前体细胞　；参考：《南昌大学》2014年硕士论文

【摘要】：目的：采用G93A-SOD1转基因小鼠作为肌萎缩侧索硬化（ALS）动物模型，观察分析正常生理条件下和ALS病理条件下成年小鼠脊髓神经前体细胞（NPCs）的分布、增殖、迁移途径和分化方向以及HoxB9的表达情况。方法：构建B6SJL-Tg（SOD1*G93A）1Gur/NJU转基因小鼠动物模型，用PCR技术检测和筛选转基因小鼠动物模型。采用荧光免疫组织化学双标记技术，分别标记NPCs、神经元细胞、星形胶质细胞、少突胶质细胞、HoxB9基因表达的细胞。每一解剖区同时进行双重荧光免疫组织化学染色，在显微镜下观察，并用图像处理技术将不同染色的阳性细胞两两叠加成像，观察分析阳性细胞分布特征，，计算各种阳性细胞数。并统计分析相关解剖区NPCs、神经元细胞和神经胶质细胞的数量及HoxB基因家族表达细胞类型和表达量的变化。结果： 1、正常成年小鼠脊髓组织内存在NPCs，并主要分布于中央管周围区域，而在灰质区域内少量散在分布。不同节段间中央管周围区域的NPCs在数量上存在颈段＞腰段＞胸段的规律。 2、在ALS病理状态下发病后的小鼠脊髓内存在NPCs的大量增殖，且在各个解剖区域均呈现出进展组＞起病组＞未发病组的规律。 3、在ALS病理状态下小鼠脊髓灰质（灰质前角、后角、侧角区域）内增殖的NPCs与Anti-GFAP antibody存在共免疫反应性。提示增殖的NPCs主要分化为星形胶质细胞。 4、在正常和ALS病理状态下的小鼠脊髓内均不存在HoxB9基因表达，提示在成年小鼠脊髓内HoxB9基因已不再表达，且在ALS病理条件下不能激活其表达。结论： 1、在正常生理条件下成年小鼠脊髓内NPCs主要分布于中央管周围区域内，而在ALS病理状态下，可刺激成年小鼠脊髓内NPCs随疾病的进展大量增殖并向着损伤区域迁移，并且灰质区域内增殖的NPCs大多分化为星形胶质细胞。 2、HoxB9基因在正常及ALS成年小鼠脊髓内均不再表达。
[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.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R744.8

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