Upgrade of fluorescence microscopy imaging system.

这个应用程序是一个数码相机和软件，以支持我的NSERC DG研究项目，目前专注于一种允许镁和钙被细胞吸收的通道蛋白的特征和作用。这种蛋白质，瞬时受体潜力Melastatin(TRPM)通道，异构体7，存在于各种各样的细胞中。它在结构上是独一无二的，因为它既包括一个跨细胞膜移动离子的通道，也包括一个用于磷酸化从而改变许多其他蛋白质活性的激酶。虽然已经知道TRPM7是细胞分裂和分化所必需的，但通道和激酶活性在这些重要的生物学过程中的作用还不清楚。我们已经在实验室建立了一系列的细胞系和成年肝细胞，代表了从胚胎到未分裂的成年细胞的肝细胞分化以及肝癌的去分化。随着成像设备和膜片钳技术的结合使用，我们最近发现，向成体分化与细胞表面的通道活性降低和围绕细胞核的膜中通道蛋白的增加有关。此外，在细胞分裂过程中，TRPM7与有丝分裂纺锤体相关，暗示着细胞外围的通道功能之外的额外作用。成像/分析升级将使我们能够继续研究TRPM7在分化肝母细胞、分裂肝癌细胞和未分裂的成年肝细胞中的分子特征和生理作用。这项工作将进一步深入了解TRPM7的一般细胞生物学，特别是正常肝脏发育所必需的事件。这一结果将有助于我们进一步了解癌细胞的不受控制的生长，更具体地说，是从越来越常见的脂肪肝和肝炎到肝癌的肝病进展。此外，相机和软件将允许我继续在我的实验室培训学生和技术人员，了解最先进的方法来理解肝细胞生理学的分子过程。

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.