Drosophila CNS OXPHOS inhibition microarray

Mitochondrial dysfunction causes biophysical, metabolic and signalling changes that alter homeostasis and reprogram cells. We used a Drosophila model in which individual subunits of 4 OXPHOS complexes are knocked-down in neurons in the larval CNS. We used microarray analysis to investigate gene expression changes caused by knock down of OXPHOS subunits of complexes I, III, IV and V in neurons. RNA was purified from the late third instar larval CNS from control larvae (nSyb-Gal4/w), or larvae expressing RNAi transgenes targeting ND-75 (complex I, RNAi line #33910), UQCR-14 (complex III, RNAi line #109542), COX5B (complex IV, RNAi line #105769) and ATPsynCf6 (complex V, RNAi line #107826) in neurons using nSyb-Gal4. Three replicates are included for the each condition.

线粒体功能异常可引发生物物理、代谢与信号转导层面的改变，进而扰乱细胞稳态并介导细胞重编程。本研究采用果蝇（Drosophila）模型，该模型中幼虫中枢神经系统（central nervous system, CNS）的神经元内，4种氧化磷酸化（OXPHOS）复合物的单个亚基被敲低。我们通过微阵列（microarray）分析，探究了神经元内氧化磷酸化复合物I、III、IV及V的亚基被敲低后所诱导的基因表达变化。实验样本取自三龄晚期幼虫的中枢神经系统，分为对照组与实验组：对照组幼虫的基因型为nSyb-Gal4/w；实验组幼虫则通过神经元特异性Gal4驱动系nSyb-Gal4，分别表达靶向ND-75（复合物I，RNA干扰[RNAi]品系#33910）、UQCR-14（复合物III，RNAi品系#109542）、COX5B（复合物IV，RNAi品系#105769）及ATPsynCf6（复合物V，RNAi品系#107826）的RNA干扰转基因片段。每种实验条件均设置三次生物学重复。