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

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，该研究在很大程度上依赖于对特定大脑区域或细胞群中大量基因表达的准确定量。这种类型的信息可以告诉我们特定神经元通路的活动以及涉及行为，认知，发育和衰老的大脑过程的机制。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品上工作，并且提供非常准确的测量。申请的设备将使申请人能够面对其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，目前的研究包括许多情况下，样本是微量的，例如当申请人解剖大脑的一小部分时，或者当他们研究大脑中存在的非常小数量的非常特定的神经元群体时。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，非常适合这种棘手的样品。我们希望将这种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通路的基因表达。步行者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源（例如妊娠并发症或孕产妇护理）对幼崽后期大脑功能和行为的影响。弗洛雷斯博士将确定参与大脑发育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，并允许大量的研究生和本科生的多样化神经科学培训。

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.