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

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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.