Establishing a canine brain and tissue bank – molecular validation by RT-qPCR targeting three reference genes

Background: Dogs ( Canis familiaris ) are natural models of several human diseases, including age-related dementia. However, the molecular techniques, which are routinely applied in invertebrate and rodent models to study disease pathologies and mechanisms, has limited applicability in dogs, mainly because of ethical reasons. In the case of humans, the limited accessibility of tissue samples is at least partly solved by biobanks, which collect and store tissues and organs from voluntary donations. A similar approach with pet dogs could support both translational and veterinary research goals by providing access to good quality biological materials obtained from a wide range of dogs with known ancestry, life history and medical background. Therefore, we have established an initiative, the Canine Brain and Tissue Bank, to collect and store biological samples from pet dogs. Objectives: The molecular qualities of tissue samples collected and stored on a tissue bank are crucial for reliable downstream applications. We used quantitative Real-Time PCR methodology to assess the stability of mRNA content in our stored samples. Three previously validated reference genes, GAPDH , HMBS and HPRT1 were chosen as targets. Results: The tested reference genes showed expression patterns and stability values that were consistent with the literature. Conclusions: Based on the results, the molecular quality of tissues collected in the CBTB’s donations system can fit in the standards of dog gene expression analyses.