Transcriptomic analysis of the hypothalamic paraventricular and the arcuate nucleus of NNAT null mice

Neuronatin (Nnat) has previously been reported to be part of a network of imprinted genes. Disruption of neuronatin results in an unusual phenotype of a bimodal body weight.In order to understand at the molecular level how the Nnat deficiency could contribute to the hypervariable phenotypes seen, we performed RNA sequencing from laser-capture micro dissected paraventricular nucleus (PVN) and arcuate nucleus (ARC) and compared the transcriptome between Nnat+/-p mice and their Nnat+/+ littermates in different feeding conditions. We performed an analysis of three subgroups: Nnat+/+ (all non-obese), Nnat+/-p non-obese, and Nnat+/-p obese mice and combined two approaches to identify differences in hypothalamic gene expression between subgroups Overall design: Gene expression analysis between wild-type and NNAT null mice神经连蛋白（Neuronatin, Nnat）此前已被证实属于印记基因（imprinted genes）调控网络的组成成员。神经连蛋白的功能缺失会引发一种特殊的双峰体重表型。为从分子层面解析Nnat缺陷如何导致所观测到的高变异性表型，我们对经激光捕获显微切割（laser-capture microdissection）获取的下丘脑室旁核（paraventricular nucleus, PVN）与弓状核（arcuate nucleus, ARC）开展了RNA测序（RNA sequencing），并在不同喂养条件下，对比了Nnat+/-p小鼠与其野生型同窝仔鼠（Nnat+/+）的转录组（transcriptome）差异。 本研究设置了三个分析亚组：全部为非肥胖个体的Nnat+/+组、Nnat+/-p非肥胖组，以及Nnat+/-p肥胖组，并结合两种分析策略，以识别不同亚组间下丘脑基因表达的差异。 整体实验设计：野生型（wild-type）小鼠与NNAT敲除小鼠（null mice）的基因表达分析