大鼠外周神经节内神经前体细胞的观察
发布时间：2019-04-11 18:03
【摘要】： 第一部分不同年龄大鼠三叉神经节、颈交感神经节、脊髓背根神经节内神经前体细胞的观察 目的观察胚胎、新生和成年大鼠三叉神经节、颈交感神经节和脊髓背根神经节内是否存在神经前体细胞。方法实验动物经灌流后,取胚胎、新生和成年大鼠三叉神经节、颈交感神经节和脊髓背根神经节行冰冻切片,进行p75,Nestin免疫组织化学和Brdu/ Nestin,Brdu/ p75免疫荧光组织化学检测。另取上述神经节进行原代培养和传代培养,观察其体外生长情况。以胎牛血清对传代培养细胞进行诱导分化。对原代细胞进行Nestin免疫组织化学和Brdu免疫荧光鉴定;对传代诱导细胞进行NF、GFAP免疫荧光鉴定。结果在胚胎、新生、成年大鼠的三叉神经节、脊髓背根神经节内均见有Brdu/ Nestin,Brdu/ p75免疫荧光双标细胞。取自胚胎和新生大鼠三叉神经节和脊髓背根神经节的细胞经原代培养,呈现Nestin和Brdu免疫荧光阳性。培养第三代细胞加入胎牛血清后出现分化,呈现NF、GFAP免疫荧光阳性。结论:在胚胎、新生、成年大鼠的三叉神经节、脊髓背根神经节和成年大鼠的颈交感神经节中存在有增殖分裂的神经前体细胞,这些细胞可以分化为神经元和神经胶质细胞。 第二部分疼痛刺激对成年大鼠脊髓背根神经节内神经前体细胞增殖与分化的影响 目的评价疼痛刺激是否会激发成年大鼠的脊髓背根神经节内的神经前体细胞的增殖与分化。方法用1%福尔马林注射于大鼠左前掌制造疼痛模型,将疼痛模型分为3天组和7天组,各4只,另设正常对照组4只。3组皆连续给予Brdu标记3天, 3天后将动物分别于不同时间点处死。对正常对照组和模型3天组大鼠的背根节进行Brdu/ Nestin免疫荧光双标检测,比较疼痛通路上的背根节内,与非疼痛通路上的背根节和正常组的大鼠背根节内Brdu/ Nestin免疫荧光双标细胞数目。模型7天组的疼痛通路上背根节与非疼痛通路上背根节行BrdU/NF、BrdU/GFAP免疫荧光双标检测,比较各组双标细胞数。结果3组大鼠各节段背根节内都可见到Brdu/ Nestin双标细胞,但疼痛通路上的双标细胞数量并未见与非疼痛通路和正常组的双标细胞数量有明显差别。7天组背根节内的BrdU/GFAP双标细胞数量多于BrdU/NF双标细胞。结论疼痛刺激对成年大鼠背根节内神经前体细胞的增殖作用并不明显,成年大鼠背根节内增殖的前体细胞在疼痛刺激的作用下主要分化为神经胶质细胞。
[Abstract]:Part one: observation of neural precursor cells in trigeminal ganglion, cervical sympathetic ganglion and dorsal root ganglion of spinal cord in rats of different ages objective to observe the trigeminal ganglion of embryonic, newborn and adult rats, Whether neural progenitor cells exist in cervical sympathetic ganglia and dorsal root ganglion of spinal cord. Methods Frozen sections of trigeminal ganglion, cervical sympathetic ganglia and dorsal root ganglia of spinal cord were obtained from embryonic, newborn and adult rats after perfusion. P75, nestin immunohistochemistry and Brdu/ Nestin, were performed. Brdu/ p75 immunofluorescence histochemistry. In addition, the primary culture and subculture of the above ganglia were performed to observe the growth of the ganglia in vitro. Fetal bovine serum was used to induce the differentiation of subcultured cells. Primary cells were identified by Nestin immunohistochemistry and Brdu immunofluorescence, and subcultured cells were identified by NF,GFAP immunofluorescence. Results Brdu/ Nestin,Brdu/ p75 immunofluorescence double labeled cells were found in trigeminal ganglion and dorsal root ganglia of embryonic, neonatal and adult rats. The cells from trigeminal ganglion and spinal dorsal root ganglion of embryonic and neonatal rats were primarily cultured and showed positive immunofluorescence of Nestin and Brdu. The third generation cells were differentiated with fetal bovine serum and showed NF,GFAP immunofluorescence positive. Conclusion: in trigeminal ganglion of embryonic newborn adult rat dorsal root ganglion of spinal cord and cervical sympathetic ganglia of adult rat there are proliferating and mitotic neural progenitor cells which can differentiate into neurons and glial cells. Part two effect of pain stimulation on proliferation and differentiation of neural precursor cells in spinal dorsal root ganglion of adult rats objective to evaluate whether pain stimulation can stimulate the spinal dorsal root nerve of adult rats Proliferation and differentiation of neural precursor cells. Methods the pain model was made by injecting 1% formalin into the left anterior metacarpal of rats. The pain model was divided into 3-day group and 7-day group with 4 rats in each group, and 4 rats in the normal control group. All of the 3 groups were labeled with Brdu for 3 days. The animals were killed at different time points after 3 days. The dorsal root ganglia of normal control group and model 3 day group were detected by Brdu/ Nestin immunofluorescence double labeling, and the pain pathway in the dorsal root ganglia was compared. The number of Brdu/ Nestin immunofluorescence double labeled cells in dorsal root ganglia with non-pain pathway and normal rats. The dorsal root ganglia on pain pathway and the dorsal root node on non-pain pathway in the 7-day model group were detected by BrdU/NF,BrdU/GFAP immunofluorescence double labeling, and the number of double-labeled cells in each group was compared. Results double labeled Brdu/ Nestin cells were observed in the dorsal root ganglia of the three groups. However, the number of double labeled cells in the pain pathway was not significantly different from that in the non-pain pathway and the normal group. The number of BrdU/GFAP double-labeled cells in the dorsal root ganglion of the 7-day group was higher than that of the BrdU/NF double-labeled cells. Conclusion the effect of pain stimulation on the proliferation of neural progenitor cells in the dorsal root ganglion of adult rats is not obvious. The neural progenitor cells proliferated in the dorsal root ganglion of adult rats are mainly differentiated into glial cells under the action of pain stimulation.
【学位授予单位】：东南大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R329
本文编号：2456639
[Abstract]:Part one: observation of neural precursor cells in trigeminal ganglion, cervical sympathetic ganglion and dorsal root ganglion of spinal cord in rats of different ages objective to observe the trigeminal ganglion of embryonic, newborn and adult rats, Whether neural progenitor cells exist in cervical sympathetic ganglia and dorsal root ganglion of spinal cord. Methods Frozen sections of trigeminal ganglion, cervical sympathetic ganglia and dorsal root ganglia of spinal cord were obtained from embryonic, newborn and adult rats after perfusion. P75, nestin immunohistochemistry and Brdu/ Nestin, were performed. Brdu/ p75 immunofluorescence histochemistry. In addition, the primary culture and subculture of the above ganglia were performed to observe the growth of the ganglia in vitro. Fetal bovine serum was used to induce the differentiation of subcultured cells. Primary cells were identified by Nestin immunohistochemistry and Brdu immunofluorescence, and subcultured cells were identified by NF,GFAP immunofluorescence. Results Brdu/ Nestin,Brdu/ p75 immunofluorescence double labeled cells were found in trigeminal ganglion and dorsal root ganglia of embryonic, neonatal and adult rats. The cells from trigeminal ganglion and spinal dorsal root ganglion of embryonic and neonatal rats were primarily cultured and showed positive immunofluorescence of Nestin and Brdu. The third generation cells were differentiated with fetal bovine serum and showed NF,GFAP immunofluorescence positive. Conclusion: in trigeminal ganglion of embryonic newborn adult rat dorsal root ganglion of spinal cord and cervical sympathetic ganglia of adult rat there are proliferating and mitotic neural progenitor cells which can differentiate into neurons and glial cells. Part two effect of pain stimulation on proliferation and differentiation of neural precursor cells in spinal dorsal root ganglion of adult rats objective to evaluate whether pain stimulation can stimulate the spinal dorsal root nerve of adult rats Proliferation and differentiation of neural precursor cells. Methods the pain model was made by injecting 1% formalin into the left anterior metacarpal of rats. The pain model was divided into 3-day group and 7-day group with 4 rats in each group, and 4 rats in the normal control group. All of the 3 groups were labeled with Brdu for 3 days. The animals were killed at different time points after 3 days. The dorsal root ganglia of normal control group and model 3 day group were detected by Brdu/ Nestin immunofluorescence double labeling, and the pain pathway in the dorsal root ganglia was compared. The number of Brdu/ Nestin immunofluorescence double labeled cells in dorsal root ganglia with non-pain pathway and normal rats. The dorsal root ganglia on pain pathway and the dorsal root node on non-pain pathway in the 7-day model group were detected by BrdU/NF,BrdU/GFAP immunofluorescence double labeling, and the number of double-labeled cells in each group was compared. Results double labeled Brdu/ Nestin cells were observed in the dorsal root ganglia of the three groups. However, the number of double labeled cells in the pain pathway was not significantly different from that in the non-pain pathway and the normal group. The number of BrdU/GFAP double-labeled cells in the dorsal root ganglion of the 7-day group was higher than that of the BrdU/NF double-labeled cells. Conclusion the effect of pain stimulation on the proliferation of neural progenitor cells in the dorsal root ganglion of adult rats is not obvious. The neural progenitor cells proliferated in the dorsal root ganglion of adult rats are mainly differentiated into glial cells under the action of pain stimulation.
【学位授予单位】：东南大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R329
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