EXPRESSION OF PROTEIN KINASE C IN LENS EPITHELIAL CELLS

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Changes in protein kinase C (PKC) isoform expression has been demonstrated in various diabetic tissues. This has been also shown with cells exposed to high concentrations of sugars as a system to study diabetes. Using Western blot analysis the P.I. determined that in bovine lens epithelial cells in culture exposure to high concentration of galactose (40 mM) resulted in changes in the relative amounts of one of the major PKC isoforms when compared to cells grown in normal (low sugar) media. Changes in the cellular localization of two of the major PKC isoforms, PKC-alpha and PKC-gamma, was also observed when comparing normal and galactosemic bovine lens epithelial cells in culture using epifluorescent microscopy. The aim of this project is to determine if changes in PKC isoforms occur at the RNA level in lens epithelial cells in culture after exposure to 40 mM galactose. This will be accomplished by establishing primary cultures of bovine lens epithelial cells and using techniques such as Northern blot analysis to determine if the expression of the PKC isoforms changes. In situ hybridization will be used to determine the expression pattern of the PKC transcripts. Expression will be studied using the appropriate PKC isoform specific cDNA probes. The findings obtained through this project will provide information for the elucidation of the mechanism of development of galactosemic cataracts. Another aim of this project is to increase the number of minority students performing research at the institution. Enhancing the awareness of our students to continue graduate studies in Cellular and Molecular Biology will also be assessed.