Upgrade of fluorescence microscopy imaging system.

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This application is for a digital camera and software to support my NSERC DG research program, currently focused on the characterization and roles of a channel protein that permits magnesium and calcium to be absorbed by cells. This protein, Transient Receptor Potential Melastatin (TRPM) channel, isoform 7, exists in a wide variety of cells. It is structurally unique in that it consists of both a channel for moving ions across cell membranes, and a kinase, for phosphorylating and hence modifying the activity of many other proteins. Although it is known that TRPM7 is required for cell division and differentiation from embryonic to adult cell type, the contribution of channel and kinase activities to these vital biological processes is not understood. We have established in the lab a series of cell lines and adult hepatocytes that represent a spectrum of liver cell differentiation from embryonic to non-dividing adult cells and de-differentiation of liver cancer. With the use of imaging equipment in tandem with the patch clamp technique, which allows us to measure ion conducting activity of the channels in live cells, we recently showed that differentiation to the adult form is associated with decreased channel activity at the surface of the cell and increased channel protein in the membrane encircling the nucleus. Further, during cell division TRPM7 was associated with the mitotic spindle, implicating roles additional to channel function at the cell periphery. The imaging/analysis upgrade will allow us to continue our studies into the molecular characterization and physiological roles of TRPM7 in differentiating hepatoblasts, dividing hepatoma cells and non-dividing adult hepatocytes. This work will provide further insight into the cell biology of TRPM7 in general, and specifically to the events necessary for normal liver development. The results will be important in furthering our understanding of the uncontrolled growth of cancer cells and, more specifically, liver disease progression from the more and more common fatty liver and hepatitis to liver cancer. Further, the camera and software will allow me to continue training students and technicians in my laboratory on state-of-the-art approaches to understanding the molecular processes underlying hepatocellular physiology.