猕猴转基因载体病毒的筛选和新型猕猴认知自动测试系统的建立

发布时间：2018-04-29 02:39

本文选题：阿尔茨海默病模型 + 猕猴　；参考：《中国科学技术大学》2014年硕士论文

【摘要】：阿尔茨海默病(Alzheimer's disease,D)是一种以老年斑和神经纤维缠结为主要病理特征的神经退行性脑疾病。关于AD的发病机制,两个最有影响力的假说是淀粉样蛋白(myloid β, Aβ)级联假说和tau蛋白假说。AD多起病于老年期,AD病人认知功能进行性丢失,以记忆损害为早期表现。被确诊后的AD患者,剩余的存活期平均约只有5.7年,并且患者发病后一直需要有人照顾,后期生活不能自理。AD患病人数已经超过2600万且在逐渐增多,给家庭和社会带来重大的经济和精神负担。AD至今无有效的治疗方法,而基于AD小鼠模型开发的许多药物的临床试验失败。猕猴与人类亲缘关系更近,并且在老年猕猴脑中发现存在Ap斑块。因此建立能反应AD病程的有效猕猴模型,探索AD的疾病机理,并寻找有效的诊断和治疗方法,既具有重大的科学意义,又是亟待解决的科学难题。我们实验室拟采用向猕猴大脑皮层注射携带家族性AD基因的病毒或体外合成并培养的纤维化Aβ42的方法,来建立AD猕猴模型。建模的两个关键准备工作是：1)筛选合适的病毒载体,为下一步的转基因工作做准备；2)建立一套新型认知检测系统,用来对猕猴实验处理前后的学习记忆能力进行评价。这两部分工作构成了本论文的主要内容。病毒筛选方面,根据国内外文献,我们尝试了慢病毒和腺相关病毒(AdenoAssociated Virus,AAV),总共有15种。在猕猴脑中只有AAV2和AAV2/9这两种病毒有表达,而对照实验在大鼠脑中只有携带Swedish/Dutch/London AD基因的慢病毒、AAV2/8、AAV2和AAV2/9,这四种病毒有表达,在此基础上我们还摸索出一种合适的病毒注射方法。在猕猴认知系统建立方面,我们建立了一套猕猴自由活动情况下,触摸屏认知自动综合测试新型系统。这套系统在国内属于首创。该系统实现了4种测试软件,主要用来测试猕猴空间和非空间的工作记忆能力。软件中的视觉空间学习测试,已有三只猕猴学会了,并都做满了1000个任务。此外,我们还开发了一套较好的猕猴训练方法。所有的努力为建立AD猕猴模型奠定了重要的基础。
[Abstract]:Alzheimer's disease (Alzheimer's disease) is a neurodegenerative brain disease characterized by senile plaques and neurofibrillary tangles. About the pathogenesis of AD, the two most influential hypotheses are the amyloid 尾 (A 尾) cascade hypothesis and the tau protein hypothesis. After being diagnosed with AD, the remaining survival time is only about 5.7 years on average, and the patient has been in need of care since the onset of the disease. The number of AD patients who cannot take care of themselves in later life has exceeded 26 million and is gradually increasing. A significant economic and psychological burden on families and society. There is no effective treatment for AD, and many drugs based on AD mouse models have failed in clinical trials. Rhesus monkeys are more closely related to humans, and Ap plaques are found in the brains of old rhesus monkeys. Therefore, it is of great scientific significance to establish an effective rhesus monkey model that can reflect the course of AD, to explore the disease mechanism of AD, and to find effective diagnosis and treatment methods. We intend to establish a rhesus monkey model by injecting virus carrying familial AD gene or fibrotic A 尾 42 synthesized and cultured in vitro into the cerebral cortex of rhesus mulatta. The two key preparations for modeling are: 1) screening appropriate virus vectors and preparing for the next step of transgenic work A) A new cognitive detection system is established to evaluate the learning and memory ability of rhesus monkeys before and after experimental treatment. These two parts of the work constitute the main content of this paper. In the aspect of virus screening, according to the domestic and foreign literature, we have tried lentivirus and adeno-associated virus AAVA, there are 15 species in total. Only AAV2 and AAV2/9 were expressed in the brain of rhesus monkeys, while in the control experiment, there were only two lentiviruses, AAV2 / 8 AAV2 and AAV2 / 9, which carried Swedish/Dutch/London AD gene in the brain of rats. On the basis of this, we found a suitable method of virus injection. In the aspect of rhesus monkey cognitive system, we set up a new system for the automatic test of Rhesus monkey's cognition under the condition of free movement of Rhesus mulatta (Rhesus mulatta). This system is the first in China. The system implements four kinds of testing software, mainly used to test the spatial and non-spatial working memory ability of rhesus monkey. In the software, three macaques have learned to learn the visual space and have done 1000 tasks. In addition, we have also developed a better training method for rhesus monkeys. All these efforts have laid an important foundation for the establishment of AD rhesus monkey model.
【学位授予单位】：中国科学技术大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R749.16

【参考文献】