Wang Xingjun, Zhao Yu, Hu Yujia, Ren Pu, Sun Ying, Guo Xiaowei, Huang Xirui, Zhu Yumeng, Chen Xinhong, Feng Yu, Xue Lei
Institute of Intervention Vessel, Shanghai 10 People's Hospital, Shanghai Key Laboratory of Signaling and Diseases Research, School of Life Science and Technology, Tongji University, 1239 Siping Road, Shanghai 200092, China.
J Biol Methods. 2016 Jun 7;3(2):e43. doi: 10.14440/jbm.2016.61. eCollection 2016.
Alzheimer's disease (AD) is the most common form of dementia that affects people's health greatly. Though amyloid precursor protein (APP) has been implicated in the pathogenesis of AD, the exact role of APP and its underlying mechanism in AD progression have remained largely elusive. has been extensively used as a model organism to study a wide range of human diseases including AD. In this protocol, we expressed full length human APP in the nervous system and examined its effect on locomotion and choice ability. We found that expression of APP produced locomotion defects in larvae as measured by plate crawling ability assay (PCA), and in adult flies as monitored by plate cycling ability assay (CLA). In addition, expression of APP results in male courtship choice (MCC) defect, since wild-type males court preferentially toward young virgin females over old ones, while APP-expressing males failed to show this preference. This protocol enables us to screen for novel AD candidate genes as well as therapeutic compounds to ameliorate the disease.
阿尔茨海默病(AD)是最常见的痴呆形式,对人们的健康有极大影响。尽管淀粉样前体蛋白(APP)与AD的发病机制有关,但APP在AD进展中的具体作用及其潜在机制在很大程度上仍不清楚。 已被广泛用作研究包括AD在内的多种人类疾病的模式生物。在本实验方案中,我们在神经系统中表达全长人类APP,并检测其对运动和选择能力的影响。我们发现,通过平板爬行能力测定法(PCA)测量,APP的表达在幼虫中产生运动缺陷,而通过平板循环能力测定法(CLA)监测,在成年果蝇中也产生运动缺陷。此外,APP的表达导致雄性求偶选择(MCC)缺陷,因为野生型雄性优先向年轻处女雌果蝇求偶,而不是向年老雌果蝇求偶,而表达APP的雄性则没有表现出这种偏好。本实验方案使我们能够筛选新的AD候选基因以及改善该疾病的治疗化合物。