Drosophila CNS mitochondrial DNA dysfunction microarray

Mitochondrial DNA (mtDNA) encodes essential components of the respiratory chain and loss of mtDNA leads to mitochondrial dysfunction and neurodegeneration. Mitochondrial transcription factor A (TFAM) is an essential component of mtDNA replication and a regulator of mitochondrial copy number in cells. Studies have shown that TFAM knockdown leads to mitochondrial dysfunction and respiratory chain deficiencies. ATP synthase is Complex V of the mitochondrial respiratory chain. It is driven by a proton gradient between the intermembrane space and the mitochondrial matrix and generates the majority of cellular ATP. The knockdown of coupling factor 6 (Cf6), one of the components of the proton channel F0, causes dysfunction in the complex, leading to mitochondrial dysfunction and respiratory chain deficiencies. Using gene expression analysis, we aimed to investigate the effects of mtDNA dysfunction in the CNS at the molecular level. RNA was purified from the late third instar larval CNS from control larvae, or larvae over-expressing mitochondrial transcription factor A (TFAM) in post-mitotic neurons using the neuron specific driver nsyb-Gal4. Three replicates are included for each condition. RNA was purified from the late third instar larval CNS from control larvae, or larvae expressing ATP Cf6 RNAi in post-mitotic neurons using the neuron specific driver nsyb-Gal4. Three replicates are included for each condition.

线粒体DNA（mtDNA，Mitochondrial DNA）编码呼吸链的必需组成成分，mtDNA缺失会引发线粒体功能障碍与神经退行性变。线粒体转录因子A（TFAM，Mitochondrial transcription factor A）是线粒体DNA复制的必需组分，同时可调控细胞内线粒体的拷贝数。已有研究表明，TFAM敲低会导致线粒体功能障碍及呼吸链缺陷。ATP合酶是线粒体呼吸链的复合物V，它依托膜间隙与线粒体基质之间的质子梯度驱动，合成细胞内绝大多数ATP。质子通道F0的组分之一偶联因子6（Cf6，coupling factor 6）的敲低会造成该复合物功能异常，进而引发线粒体功能障碍与呼吸链缺陷。本研究采用基因表达分析技术，旨在从分子层面探究中枢神经系统（CNS，Central Nervous System）中线粒体DNA功能异常的影响。第一组实验的RNA提取自对照幼虫，以及通过神经元特异性驱动因子nsyb-Gal4在有丝分裂后神经元中过表达TFAM的三龄晚期幼虫的中枢神经系统，每个实验条件设置3次生物学重复。第二组实验的RNA提取自对照幼虫，以及通过神经元特异性驱动因子nsyb-Gal4在有丝分裂后神经元中表达ATP合酶Cf6 RNA干扰载体的三龄晚期幼虫的中枢神经系统，每个实验条件同样设置3次生物学重复。