固始鸡胚期视网膜发育及Notch1在视网膜发育中的表达变化规律研究
发布时间：2018-08-07 13:57
【摘要】： 鸡是研究鸟类动物发育常用的模式动物。视网膜由胚胎时期间脑外壁向外突起形成,属于脑外中枢系统,常作为研究中枢神经发育和再生的模式器官。脊椎动物视网膜基本组织模式相同:两个突触层(外网层和内网层)插在三细胞核层之间(外核层、内核层和神经节细胞层)。视网膜的形态发育过程包括神经祖代细胞分化为神经元和神经胶质细胞过程,以及神经元和神经胶质细胞按照一定的空间结构构建视网膜过程。Notch-Delta信号途径是一条在胚胎发育中起调控作用的保守的途径。Notch-Delta信号系统的最主要功能就是参与胚胎的分化发育,Notch1在神经视网膜细胞发育和分化过程中表达。 本实验采用常规组织学HE染色技术、免疫组织化学技术和分子半定量技术对固始鸡胚期视网膜的发育进行研究,研究结果显示:固始鸡视网膜是典型的脊椎动物视网膜,可分为色素视网膜和神经视网膜,神经视网膜又分为7层:神经纤维层、节细胞层、内网层、内核层、外网层、外核层和视锥视杆层;神经视网膜在胚期发育过程中的变化趋势是:神经视网膜厚度从E4至E20呈现波浪形变化趋势,E4至E8逐渐增厚,E8最厚,从E8开始厚度逐渐变薄,E10与E11厚度相差较大,E14厚度最薄,到E19出现第二个高峰,E11到E20神经视网膜厚度相差不大。节细胞层、内网层、内核层、外网层、外核层于E8时形成,三核层总的变化趋势是从E8开始厚度逐渐下降,E10和E11相差较大,从E12到出壳当天,厚度总体变化相对稳定;三核层的密度总的趋势变化相同,密度相比内核层最高,其次是外核层,节细胞层密度最低。免疫组化S-P法染色结果显示:NSE最早在E12的节细胞层出现,内网层和外网层有亚层形成,Vimentin从E3开始整个胚期都存在,且从E10开始,外网层出现一条阳性条带形成原因有待进一步研究。半定量研究表明在眼部E3和E18表达量相对较低,推断E18可能作为理想的时间点研究固始鸡视网膜再生。研究还发现脑部在出壳前两天Notch1表达量较低与E3相对接近,说明脑部细胞处于祖细胞状态或干细胞较多,增殖旺盛,Notch1在脑部有维持神经细胞发生和再生的能力。
[Abstract]:Chicken is a commonly used model animal to study the development of bird animals. The retina is formed from the outer wall of the diencephalon and belongs to the extracranial central system. It is often used as a model organ to study the development and regeneration of the central nervous system. The basic pattern of retina in vertebrates is the same: two synaptic layers (outer reticulum layer and inner reticulum layer) are inserted between the three nuclear layers (outer nuclear layer, nuclear layer and ganglion cell layer). The development of retinal morphology includes the differentiation of neural progenitor cells into neurons and glial cells. And neurons and glial cells build retinal process according to certain spatial structure. Notch-Delta signaling pathway is a conservative pathway that plays a regulatory role in embryonic development. Notch-Delta signaling system is the main function of participating in embryo. Notch1 was expressed during the development and differentiation of neuroretinal cells. The development of embryonic retina of Gushi chicken was studied by routine histological HE staining, immunohistochemical technique and molecular semi-quantitative technique. The results showed that the retina of Gushi chicken was typical of vertebrate retina. It can be divided into pigmented retina and nerve retina, and neuroretina is divided into 7 layers: nerve fiber layer, ganglion cell layer, inner reticulum layer, inner reticular layer, outer reticulum layer, outer nuclear layer and optic cone rod layer. The change trend of neuroretina during embryonic development is that the thickness of nerve retina changes from E4 to E20 in a wavy shape. The thickest thickness of E8 is gradually increased from E4 to E8, and the thickness of E10 and E11 is gradually thinned from E8 to E11, and the thickness of E14 is the thinnest. The second peak at E 19, E 11 to E 20, had little difference in retinal thickness. The ganglion cell layer, inner net layer, outer net layer and outer nuclear layer were formed at E8. The total change trend of the three nuclear layers was that the thickness of E10 and E11 gradually decreased from E8 to E11, and from E12 to the day of exiting the shell, the overall thickness changed relatively stable. The density of the trinuclear layer was the highest in comparison with the inner layer, followed by the outer nuclear layer and the ganglion cell layer. The results of S-P immunohistochemical staining showed that WNSE first appeared in the ganglion cell layer of E12, and Vimentin was formed in the inner and outer layers of the reticulum, and the whole embryo stage was observed from E3 and E10. The formation of a positive band in the outer layer needs further study. Semi-quantitative study showed that the expression of E3 and E18 was relatively low in the eye. It was inferred that E18 might be used as an ideal time point to study retina regeneration in Gushi chicken. It was also found that the expression of Notch1 in the brain was relatively close to that of E3 two days before exfoliation, indicating that the brain cells were in the state of progenitor cells or stem cells, and that the proliferation of Notch1 had the ability to maintain neurogenesis and regeneration in the brain.
【学位授予单位】：河南农业大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R774.1
本文编号：2170236
[Abstract]:Chicken is a commonly used model animal to study the development of bird animals. The retina is formed from the outer wall of the diencephalon and belongs to the extracranial central system. It is often used as a model organ to study the development and regeneration of the central nervous system. The basic pattern of retina in vertebrates is the same: two synaptic layers (outer reticulum layer and inner reticulum layer) are inserted between the three nuclear layers (outer nuclear layer, nuclear layer and ganglion cell layer). The development of retinal morphology includes the differentiation of neural progenitor cells into neurons and glial cells. And neurons and glial cells build retinal process according to certain spatial structure. Notch-Delta signaling pathway is a conservative pathway that plays a regulatory role in embryonic development. Notch-Delta signaling system is the main function of participating in embryo. Notch1 was expressed during the development and differentiation of neuroretinal cells. The development of embryonic retina of Gushi chicken was studied by routine histological HE staining, immunohistochemical technique and molecular semi-quantitative technique. The results showed that the retina of Gushi chicken was typical of vertebrate retina. It can be divided into pigmented retina and nerve retina, and neuroretina is divided into 7 layers: nerve fiber layer, ganglion cell layer, inner reticulum layer, inner reticular layer, outer reticulum layer, outer nuclear layer and optic cone rod layer. The change trend of neuroretina during embryonic development is that the thickness of nerve retina changes from E4 to E20 in a wavy shape. The thickest thickness of E8 is gradually increased from E4 to E8, and the thickness of E10 and E11 is gradually thinned from E8 to E11, and the thickness of E14 is the thinnest. The second peak at E 19, E 11 to E 20, had little difference in retinal thickness. The ganglion cell layer, inner net layer, outer net layer and outer nuclear layer were formed at E8. The total change trend of the three nuclear layers was that the thickness of E10 and E11 gradually decreased from E8 to E11, and from E12 to the day of exiting the shell, the overall thickness changed relatively stable. The density of the trinuclear layer was the highest in comparison with the inner layer, followed by the outer nuclear layer and the ganglion cell layer. The results of S-P immunohistochemical staining showed that WNSE first appeared in the ganglion cell layer of E12, and Vimentin was formed in the inner and outer layers of the reticulum, and the whole embryo stage was observed from E3 and E10. The formation of a positive band in the outer layer needs further study. Semi-quantitative study showed that the expression of E3 and E18 was relatively low in the eye. It was inferred that E18 might be used as an ideal time point to study retina regeneration in Gushi chicken. It was also found that the expression of Notch1 in the brain was relatively close to that of E3 two days before exfoliation, indicating that the brain cells were in the state of progenitor cells or stem cells, and that the proliferation of Notch1 had the ability to maintain neurogenesis and regeneration in the brain.
【学位授予单位】：河南农业大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R774.1
【参考文献】
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1 殷咏仪,华茜;视网膜发育与形成的“镶嵌模型”[J];生物化学与生物物理进展;2001年02期
，本文编号：2170236
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