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摘要帕金森病(PD)是一种大脑(中枢神经系统),经常损害运动技能、语言和其他功能的退化性疾病。PD长期被认为是由环境因素引起的,但发现的几个基因突变的患者(主要是家庭形式的PD)清楚地证明了遗传因素参与帕金森病的发展。在这些确定的基因之间,LRRK2被发现是对PD的最重要的遗传因素之一。然而,生物体内基因LRRK2的功能很大程度上是不为人知的。在这项研究中,通过阻断体内基因LRRK2的正常功能，我们研究了斑马鱼体内基因LRRK2的功能。斑马鱼表现出的神经变性和运动缺陷的特征,类似于帕金森病的患者。鱼的缺陷可以通过表达LRRK2的正常蛋白质救出，并且运动缺陷也可以通过通常用于治疗帕金森病患者施用的L-多巴救出。因此,我们已经开发出一种PD斑马鱼模型可用于理解PD的发展机制,将有助于未来治疗帕金森病新药的筛选。58519
LRRK2在帕金森病(PD)扮演着一个重要的角色,但其生物功能在很大程度上是未知的。在这里,我们克隆人类LRRK2的同源物,其特征在于它的表达,研究其在斑马鱼的生物功能。通过吗啉，体内基因LRRK2斑马鱼(zLRRK2)蛋白质的堵塞引起胚胎死亡率和严重发育缺陷,如生长迟缓和神经元的损失。
通过RNA-seq技术，把野生型和突变型的斑马鱼基因序列测出来进行比较，反映出它们的表达水平。通过野生型和突变型斑马鱼基因的对比，发现有许多基因存在着明显的差异。tnfrsf14基因在突变型和野生型斑马鱼中差异较为明显，所以我们要对其进行探究。
原位杂交是指将特定标记的已知顺序核酸为探针与细胞或组织切片中核酸进行杂交，从而对特定核酸顺序进行精确定量定位的过程。我们要在野生型和突变型斑马鱼中用tnfrsf14的探针进行原位杂交。
Abstract Parkinson's disease (PD) is a brain (central nervous system), often impair motor skills, language, and other features of degenerative diseases. PD has long been thought to be caused by environmental factors, but several mutations found in patients (mainly family forms PD) clearly demonstrate the genetic factors involved in the development of Parkinson's disease. In between these identified genes, LRRK2 was found to be one of the most important genetic factors of PD. However, in vivo gene LRRK2 function is largely unknown. In this study, by blocking the normal function of LRRK2 gene in vivo, we studied the zebrafish gene LRRK2 function. Zebrafish exhibited neurodegenerative and motor deficits characteristic, similar to patients with Parkinson's disease. Fish defects can express the normal protein LRRK2 rescued, and motor deficits can also commonly used to treat Parkinson's patients administered L- dopa rescued. Therefore, we have developed a PD zebrafish PD model can be used to understand the development of mechanisms that will help the future screening of new drugs to treat Parkinson's disease.
LRRK2 in Parkinson's disease (PD) plays an important role, but its biological function is largely unknown. Here, we cloned the human homologue of LRRK2, characterized in that it expressed, to study its biological function in zebrafish. By morpholine, LRRK2 gene in vivo zebrafish (zLRRK2) protein blockage caused embryonic mortality and severe developmental defects, such as growth retardation and loss of neurons.
By RNA-seq technology, the wild-type and mutant zebrafish gene sequences to compare measured out, reflecting their expression levels. By comparing wild-type and mutant zebrafish gene, we found that many genes there are obvious differences. tnfrsf14 gene in mutant and wild-type zebrafish difference more obvious, so we have to be explored.
In situ hybridization refers to a known sequence specific nucleic acid probe labeled with a cell or tissue sections nucleic acid hybridization to specific nucleic acid sequence and thus the course of precise quantitative targeting. We want to in the wild type and mutant zebrafish with tnfrsf14 probe in situ hybridization.