利用定位UAS-RNAi系统筛选果蝇运动抗心功能増龄衰退候选钙调控基因
Screening Ca2+ Regulation Genes as Candidate Genes of Exercise Anti-Cardiac Aging in Drosophila by Regional UAS-RNAi System

摘要 目的：研究果蝇心肌细胞Ca2+调控相关基因敲减对増龄果蝇心脏功能及寿命的影响，筛选与心脏功能密切相关的Ca2+调控基因作为候选基因，为研究増龄果蝇基于Ca2+调控基因功能改变的心脏运动适应机制提供依据。方法：Hand-GAL4品系雄蝇、Ca2+调控相关基因UAS-RNAi品系雄蝇与野生型W1118处女蝇杂交，获得基因型为Hand/+和calcium regulation gene RNAi/+的F1代果蝇作为对照组果蝇；Hand-GAL4品系雄蝇与Ca2+调控相关基因UAS-RNAi品系处女蝇杂交，获得F1代基因型为Hand> calcium regulation gene RNAi的果蝇为心肌组织Ca2+调控基因RNAi组果蝇。采用M-mode方法对1周龄（约为人类青少年时期）和5周龄（约为人类中老年时期）成体果蝇心脏功能衰退相关FS、AI、FL等指标进行检测，并统计各组果蝇存活情况，制作存活曲线。结果：CaMKII基因心肌敲减果蝇与对照组相比：存活率明显下降，1周龄时表现为FS下降显著，FL明显增高，5周龄时表现为DI，FS明显下降，SI，AI和FL显著上升；5周龄Ca-α1D基因心肌敲减果蝇对比同龄对照组，DI、SI明显降低，DD、SD明显增高；5周龄AnxB9基因敲减果蝇AI较同龄对照组显著上升。结论：CaMKII基因在维持増龄果蝇心脏功能和寿命中发挥着重要作用，CaMKII、Ca-α1D和AnxB9可作为候选基因研究増龄果蝇心脏功能运动适应的潜在机制。
Abstract: Objective: The purposes of this study were to explore the effects of cardiac-specific Ca2+ regulation genes knockout on cardiac function and lifespan in aging drosophila, screen Ca2+ regulation genes as candidate genes of exercise anti-cardiac aging, and provide references for the mechanism of cardiac function adapting to exercise based on Ca2+ regulation genes in aging drosophila. Methods: Male Hand-Cal4 and UAS-RNAi drosophila crossed with wild-type strain w1118 females respectively, and their filial generations were Hand/+, calcium regulation gene RNAi/+, and were collected as control group. Hand-Cal4 males and UAS-RNAi females were crossed and their filial generations were Hand> calcium regulation gene RNAi, which were collected as cardiac specific Ca2+ regulation gene knockout group. Heart function indicators, FS, AI and FL, of young drosophila (1-week old) or middle-age and old drosophila (5-week old) were examined by M-mode methods, and the survival rate of drosophila was recorded. Results: Compared with control group, survival rate of CaMKII gene knockout group decreased significantly, FS of 1-week old drosophila decreased while FL increased, DI and FS of 5-week old drosophila decreased while their SI, AI and FL increased. Compared with control group with the same age, DI and SI of 5-week old drosophila with Ca-alD knockout decreased significantly while DD and SD increased, AI of 5-week old drosophila with AnxB9 knockout increased. Conclusion: CaMKII plays an important role in heart function of aging drosophila and lifespan. CaMKII, Ca-α1D and AnxB9 can be candidate genes for the mechanism of cardiac function adapting to exercise in aging drosophila