模拟微重力环境作用于神经前体细胞分化的生物学效应与分子机制
文献类型：学位论文
| 作者 | 杨履瑜
|
| 答辩日期 | 2023-06 |
| 文献子类 | 硕士 |
| 授予单位 | 中国科学院大学 |
| 授予地点 | 中国科学院心理研究所 |
| 其他责任者 | 赵媚 |
| 关键词 | 模拟微重力 人神经前体细胞 单细胞转录组测序 人诱导多能干细胞 |
| 学位名称 | 理学硕士 |
| 学位专业 | 健康心理学 |
| 其他题名 | Effects of Simulated Microgravitv on Neural Progenitor Cells Differentiation |
| 中文摘要 | Long-term exposure to microgravity has not only affected the cardiovascular system, skeletal muscle system, and immune system of astronauts but changed the structure of the nervous system and cognitive function. Human Neural Progenitor Cells (hNPCs) are the basis of the central nervous system involved in the development of the nervous system and the occurrence of neuropsychiatric diseases. Therefore, the study of the biological effects of simulated microgravity on hNPCs differentiation and analysis of the molecular mechanism of simulated microgravity on neuronal development, so as provide an effective way to explore the mechanism of microgravity causing neurological disorders. In this study, the forebrain hNPCs derived from human induced pluripotent stem cells (hiPSCs) were differentiated into cortical neurons under simulated microgravity, to mimic the development of the embryonic neocortex and investigate the changes of neuronal types, morphology, synaptic proteins, and functional activity. Effects of microgravity on specific neuronal clusters and gene expression were analyzed by single-cell transcriptome sequencing to explore the molecular mechanism of simulated microgravity on human neural development. Morphological staining showed that simulated microgravity increased the morphological complexity, decreased the expression of BRN2, a marker of the superficial cortex, and increased the expression of Synapsinlin differentiated neurons. Further analysis showed that the expression of inhibitory neuron marker VGAT1 increased, while the expression of excitatory neuron marker VGLUTI had no effect, suggesting that simulated microgravity may promote the development of inhibitory neurons. Calcium imaging analysis showed that simulated microgravity enhanced the activity of neurons. Single-cell transcriptome analysis showed that the proportion of neurons was increased compared with neural progenitor cells, and the genes related to neural differentiation were upregulated under simulated microgravity. Besides, the increased expression of genes related to the development of neurons, synaptic plasticity (MEF2C, PSD3), and neurotrophic factor pathway (VGF, NTRK2), which may be the potential molecular mechanism of microgravity on neural differentiation. In conclusion, simulated microgravity promoted the differentiation of hiPSCs-derived hNPCs towards neurons, especially inhibitory neurons. In addition, simulated microgravity increased the expression of synaptic proteins and improved the functional activity of neurons. The significantly differentially expressed genes such as MEF2C, PSD3, and VGF may be the key molecules that simulated microgravity to regulate the differentiation of neural progenitor cells and synaptic plasticity. Understanding the biological effect of simulated microgravity on neural progenitor cell differentiation and its mechanism will provide new ideas for stem cell therapy of neuropsychiatric diseases. |
| 英文摘要 | 长期处于微重力环境，除了对宇航员的心血管系统、骨骼肌系统、免疫系统产生影响，也改变神经系统的结构与认知功能。人神经前体细胞(human Neural Progenitor Cells hNPCs)作为中枢神经系统产生的基础，参与神经系统的发育和神经精神疾病的发生过程。因此研究模拟微重力对hNPCs分化的生物学效应及影响神经元发育的分子机制，是探究微重力引起神经障碍的有效途径。 本课题利用人诱导多能干细胞(human induced Pluripotent Stem Cells hiPSCs)来源的前脑hNPCs，在模拟微重力环境下，诱导分化为大脑皮层神经元，模拟胚胎大脑新皮层发育过程，探究神经元的类型，神经元形态，突触蛋白与神经元功能活性的变化;利用单细胞转录组测序分析模拟微重力对特定神经元群及基因表达的影响，探究模拟微重力对神经发育过程的影响及分子机制。 通过形态学染色发现模拟微重力增加神经元形态的复杂性，使大脑皮层浅层神经元分子标记物B RN2表达减少，突触蛋白Synapsinl表达增加;进一步分析发现抑制性神经元标记物VGAT1表达增加，而兴奋性神经元标记物V GLUT 1的表达无影响，说明模拟微重力可能促进hNPCs向抑制性神经元的方向发育;钙成像分析表明模拟微重力增加神经元活动性;单细胞转录组分析发现模拟微重力组中，神经元相对于神经前体细胞的比例增加，促进神经分化的基因表达上调。此外，在基因表达差异最显著的抑制性神经元群1中发现与神经元发育(MEF2C),突触可塑性(PSD3)相关的基因表达上调;最重要地是，神经营养因子通路(VGF,NTRK2)相关的基因表达增加，这可能是模拟微重力作用于神经发育的潜在分子机制。 综上所述，模拟微重力环境促进hiPSCs来源的hNPCs朝神经元方向分化，尤其是抑制性神经元;此外，模拟微重力增加突触蛋白的表达，提高神经元的功能活性;MEF2C, PSD3, VGF等差异表达显著的基因可能是模拟微重力调节神经前体细胞分化及突触可塑性的关键分子;理解模拟微重力对神经前体细胞分化的生物学效应及其机制将为神经精神疾病的干细胞疗法提供新思路。 |
| 语种 | 中文 |
| 源URL | [http://ir.psych.ac.cn/handle/311026/46099]  |
| 专题 | 心理研究所_健康与遗传心理学研究室 |
| 推荐引用方式 GB/T 7714 | 杨履瑜. 模拟微重力环境作用于神经前体细胞分化的生物学效应与分子机制[D]. 中国科学院心理研究所. 中国科学院大学. 2023. |
入库方式： OAI收割
来源：心理研究所
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