Quantitative real-time PCR system to study nutrient and gut microbiota regulation of gene expression

实时PCR分析是基因表达研究的金标准技术。它用于基因表达的定量和半定量分析，作为一种独立的技术或确认微阵列结果。在几种实验条件下评估基因表达可以研究营养素或环境因素可能影响身体健康的机制。我们将使用所提出的仪器来研究各种饮食治疗后各种小鼠和大鼠组织（但主要是肠道和大脑）中的基因表达变化。此外，我们将使用它来评估肠道驻留微生物群如何影响出生后发育期间的肠上皮基因表达，通过使用从激光捕获显微切割细胞制备的RNA。特别是，我们将研究粘蛋白在常规（具有健康的肠道微生物群）和无菌（没有肠道微生物群）小鼠中的基因表达，以评估肠道细菌在出生后肠道粘液层建立中的影响。揭示宿主基因表达的营养和肠道微生物群调节机制可能会更好地了解几种肠道和肠外疾病（如炎症性肠病和肥胖症）的病因。例如，在加拿大，170，000人患有炎症性肠病，这种疾病不存在最终治愈方法。最后，这项研究有助于制定膳食指南，以改善人类健康。

Real-time PCR analysis is a gold standard technique for gene expression studies. It is used for the quantitative and semi-quantitative analysis of gene expression either as a stand alone technique or to confirm microarray findings. Assessing gene expression in several experimental conditions allows to investigate the mechanisms by which nutrients or environmental factors may impact on body health. We will use the proposed instrument to investigate gene expression changes in various mouse and rats tissues (but mainly in intestine and brain) following  various dietary treatments. Moreover, we will use it to assess how the gut resident microbiota impact on the intestinal epithelium gene expression during postnatal development, by using RNA prepared form laser capture microdissected cells. In particular, we will investigate gene expression of mucins in conventional (with a healthy gut microbiota) and germ-free (without a gut microbiota) mice to assess the impact of gut bacteria in the postnatal establishment of the intestinal mucus layer. Unraveling the mechanisms regulating nutrient and gut microbiota modulation of host gene expression may lead to a better understanding of the aetiology of several intestinal and extra-intestinal diseases such as inflammatory bowel disease and obesity. As an example, 170,000 persons suffer from inflammatory bowel disease in Canada, a disease for which a final cure does not exist. Finally, this research has implications in aiding in the development of dietary guidelines to improve human health.