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

现在，大规模和精确的基因表达测量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，该研究在很大程度上依赖于对特定大脑区域或细胞群体中大量基因表达的准确量化。这类信息可以让我们了解特定神经通路的活动，以及行为、认知、发育和衰老等大脑过程中涉及的机制。实时聚合酶链式反应技术是研究基因表达的首选方法，因为它允许对大量非常小的样本进行工作，并提供非常准确的测量。所要求的设备将允许申请者面对他们的研究计划产生的越来越多的样本，其中包括越来越复杂的实验。然而，如今的这项研究包括了许多样本数量很少的情况，例如，当申请者解剖大脑的一小部分时，或者当他们研究大脑中以非常小的数量存在的非常特定的神经元群体时。在后一种情况下，申请者将能够依靠数字聚合酶链式反应的新方法，这是本申请的另一个组成部分，它非常适合于这种棘手的样本。我们希望将这项技术应用于我们研究计划的各个方面，特别是对我们的NSERC发现基金资助的研究至关重要。Cermakian博士和Luhehi博士将分别在日常节律和炎症的背景下研究神经免疫通路背后的基因表达。沃克博士和博克萨博士将使用该设备在基因表达水平上测试早期生活应激源(例如怀孕并发症或产妇护理)对幼崽稍后生活中的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。总体而言，所要求的基因表达研究设施将极大地增强我们的研究能力，并允许对大量研究生和本科生进行多样化的神经科学培训。

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.