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Both the Chantler and Verchere laboratories are part of the Diabetes Research Program at the Child and Family Research Institute in Vancouver, British Columbia. The Diabetes Research Program includes six laboratories whose research covers aspects of the immunopathology, cell biology, biochemistry, virology, and genetics of type 1 diabetes. The focus of the Chantler laboratory is on the mechanism of pancreatic regeneration following a viral insult. In particular, the lab is using a mouse mod el to characterize growth factors and cytokines that are expressed during tissue recovery following coxsackievirus infection, using microarrays, real-time PCR, and protein expression analysis. This information is then exploited to determine whether delivering combinations of these factors to a pancreas damaged by chemical treatment, using a novel viral vector, can induce tissue recovery and normoglycemia. The focus of the Verchere laboratory is on the biology of the islet beta cell. The aims are to understand the causes of beta cell dysfunction in type 2 diabetes and the mechanisms by which beta cells are killed in both type 1 and type 2 diabetes. The goal is to develop new ways to enhance beta cell function and survival in diabetes and following islet transplantation.
www.cfri.ca/our_research/research_themes/programmatic_themes/diabetes.htm
Our research hinges on the ability to isolate intact mRNA from the pancreas of mice infected with coxsackievirus over the course of an acute infection The pancreas contains high levels of ribonucleases and isolation of intact RNA has always represented a challenge to researchers. RNA isolation using a strong denaturing agent such as guanidine hydrochloride in combination with phenol, works well for most tissues, but yields from pancreatic tissue are poor, even when the protocol includes RNase inhibitors and low temperature homogenization. The concept of delivering RNase inhibitors in vivo, before disruption of the pancreas and RNase release, arose from this challenge. In situ ductal perfusion is commonly carried out in islet isolation and is used routinely in the Verchere laboratory for islet function studies. We adapted this technique to deliver RNase inhibitors to the pancreas in vivo. The much-improved yields have enabled us to analyze pancreatic gene expression using both microarray technology and real-time PCR.
Application of in situ ductal perfusion to facilitate isolation of high-quality RNA from mouse pancreas, p. 617.
