Transcription profiling by array of ventricles from mice with homozygous knockdown expression of the vesicular acetylcholine transporter (VAChT) and wild type controls

VAChT KDHOM mice have a 70% decrease in the vesicular acetylcholine transporter (VAChT) and this leads to a systemic decrease in ACh release and cardiac dysfunction. We used this microarray to determine whether there were transcriptional alterations in the hearts of mutant mice as a result of this long-term decrease in cholinergic tone. We isolated whole hearts from 3-month-old WT and VAChT KD mice and used them for RNA extraction and hybridization on Affymetrix microarrays. We sought to obtain homogenous populations for each sample and, as such, pooled together two RNA samples to generate one microarray sample.囊泡乙酰胆碱转运体（vesicular acetylcholine transporter, VAChT）敲低纯合小鼠的VAChT表达量降低70%，该变化会引发全身性乙酰胆碱（acetylcholine, ACh）释放减少与心脏功能障碍。本研究采用该基因芯片平台，旨在探究长期胆碱能张力降低是否会导致突变小鼠心脏出现转录组学改变。我们从3月龄野生型（wild type, WT）与VAChT敲低纯合小鼠体内分离完整心脏，用于RNA提取，并在Affymetrix基因芯片上完成杂交实验。为确保每份样本的群体均一性，我们将两份RNA样本混合以制备一份芯片检测样本。