斑马鱼Lrrk2突变体中差异化表达基因Rsad2的原位杂交分析
时间:2021-11-05 20:34 来源:毕业论文 作者:毕业论文 点击:次
摘要研究表明Rsad2基因主要表达于人类大脑的神经元及神经胶质中。另外，研究发现Rsad2基因编码蛋白与先天性免疫也相关，Rsad2表达蛋白具有生物催化活性，机体Rsad2基因的缺失将导致机体对病毒免疫力下降。此外，Rsad2基因作为干扰素刺激基因，可在动物早期妊娠阶段做为胎儿产前诊断 [ ]。73727 我们先前通过RNA-seq技术手段，在分子水平上检测出斑马鱼Lrrk2突变体中的Rsad2表达量出现下调趋势。在此基础上，我们通过3dpf斑马鱼整胚原味杂交的技术手段，以WT野生型斑马鱼为实验对照，分析研究其在空间结构上的表达差异。我们从整胚原味杂交结果中发现Rsad2基因在斑马鱼大脑内的表达信号存在差异。另外，我们在研究中发现，Rsad2在早期胚胎的卵黄部位的免疫细胞上有差异化表达，通过统计分析，其卵黄部位免疫细胞个数存在明显差异，斑马鱼Lrrk2突变体卵黄部位免疫细胞个数少于WT对照组。在空间结构上，验证了3dpf斑马鱼Lrrk2突变体中Rsad2基因差异化表达，且具有明显下调趋势。 Abstract Studies show that the Rsad2 gene is mainly expressed in the neurons and glial cells of the human brain. In addition, the study found that the Rsad2 gene encoding protein is also associated with innate immunity, Rsad2 protein has a biological catalytic activity, the body of the lack of Rsad2 gene will lead to the decline of the immune system. In addition, the Rsad2 gene is used as an interferon stimulated gene, which can be used as a prenatal diagnosis in the early stage of animal pregnancy. We detected the Rsad2 expression in zebrafish Lrrk2 mutant at the molecular level by RNA-seq technique. On the basis of this, we analyzed the expression of WT in the spatial structure of 3dpf wild type zebrafish by means of the technique of whole embryo hybridization in zebrafish. We found that the expression of Rsad2 gene in the brain of zebrafish is different from the original hybridization results. Also we found in the study, Rsad2 on immune cells in the early embryo yolk parts have differential expression, through statistical analysis, the yolk immune site and the number of cells exist obvious differences, zebrafish Lrrk2 mutant yolk parts of immune cell number less than WT controls. In the level of inpidual development, the Rsad2 gene was proved to have a tendency to decrease the 3dpf gene expression in the zebrafish Lrrk2 mutant. 毕业论文关键字：斑马鱼； 整胚原位杂交； Lrrk2； Rsad2； Keywords：Zebrafish; Whole embryo in situ hybridization; Lrrk2； Rsad2； 缩略语表 缩写词 英文名称 中文名称 Confocal/LSCM laser scanning confocal microscopy 激光共聚焦显微镜 PFA Paraformaldehyde 多聚甲醛 PK Proteinase K 蛋白酶K WT Wild Type 野生型 PTU 1-phenyl 2-thiourea 1-苯基-2-硫脲 DEPC Diethy pyrocarbonate 焦碳酸二乙酯 DIG Digoxigenin 地高辛 EP Eppendorf 微量离心管 dpf Days Post Fertilization 受精后天数 hpf Hours Post Fertilization (责任编辑：qin) |