Gene expression analysis of complex neurobiological processes: A PCR facility to study large sample sets and very small samples

复杂神经生物学过程的基因表达分析：用于研究大样本集和极小样本的 PCR 设施

• 批准号: RTI-2016-00028
• 负责人: Cermakian, Nicolas
• 金额: $ 10.24万
• 依托单位: McGill University
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments
• 财政年份: 2015
• 资助国家: 加拿大
• 起止时间: 2015-01-01 至 2016-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=591313
• 关键词: Gene; expression; analysis; complex; neurobiological

The large-scale and precise measurement of gene expression is now a prerequisite to understanding complex biological mechanisms. This is particularly true in neuroscience research performed at the Douglas Institute, which relies heavily on the accurate quantification of expression of a large number of genes in specific brain areas or cell populations. This type of information can inform us on the activity of specific neuronal pathways and on the mechanisms involved in brain processes underlying behaviour, cognition, development and aging, for example. The real-time PCR technique is the method of choice to study gene expression because it allows working on large numbers of very small samples and it provides very accurate measures. The requested equipment will allow the applicants to face the increasing number of samples generated by their research program, which are including increasingly complex experiments. However, this research nowadays includes many cases where the samples are in minute quantity, for example when the applicants dissect a very small portion of the brain, or when they study a very specific neuron population which exist in very small numbers in the brain. In these latter cases, the applicants will be able to count on the new approach of digital PCR, the other component of this application, which is perfectly suited for such tricky samples. We wish to apply this technique to various aspects of our research programs, and in particular will be critical to our NSERC Discovery grant-funded research. Dr. Cermakian and Dr. Luheshi will study gene expression underlying neuro-immune pathways in the context of daily rhythms and inflammation, respectively. Dr. Walker and Dr. Boksa will use the facility to test at the gene expression level the impact of early life stressors (pregnancy complications or maternal care, for example) on brain function and behaviour later in the life of the pups. Dr. Flores will identify genetic and epigenetic events involved in the development of the brain. Overall, the requested facility for the study of gene expression will greatly enhance our research capacity and allow for the diversified neuroscience training of a large number of graduate and undergraduate students.

基因表达的大规模和精确测量现在是理解复杂生物机制的先决条件。在道格拉斯研究所进行的神经科学研究中尤其如此，该研究严重依赖于对特定大脑区域或细胞群中大量基因表达的精确量化。这类信息可以告诉我们特定神经元通路的活动，以及涉及大脑过程的机制，例如，潜在的行为、认知、发育和衰老。实时PCR技术是研究基因表达的首选方法，因为它允许在大量非常小的样本上工作，并且它提供了非常精确的测量。所要求的设备将使申请人能够面对他们的研究项目产生的越来越多的样品，其中包括越来越复杂的实验。然而，这种研究现在包括许多样品数量很少的情况，例如当申请人解剖大脑的很小一部分时，或者当他们研究大脑中数量很少的非常特定的神经元群时。在后一种情况下，申请人将能够依靠数字PCR的新方法，这是该应用程序的另一个组成部分，它非常适合这种棘手的样品。我们希望将这种技术应用到我们研究项目的各个方面，特别是对我们的NSERC发现资助的研究至关重要。Cermakian博士和Luheshi博士将分别研究日常节律和炎症背景下神经免疫途径的基因表达。沃克博士和博克萨博士将利用该设施在基因表达水平上测试早期生活压力因素（例如妊娠并发症或母性护理）对幼鼠日后大脑功能和行为的影响。弗洛雷斯博士将确定与大脑发育有关的遗传和表观遗传事件。总的来说，所要求的基因表达研究设施将大大提高我们的研究能力，并允许大量研究生和本科生进行多样化的神经科学培训。