The microvascular complications of type 1 diabetes carry a high morbidity and, when coupled with macrovascular complications, high mortality. The most common microvascular complication is neuropathy. While exact prevalence depends on the diagnostic criteria used to identify neuropathy, most studies suggest that 50% of patients with either type 1 or type 2 diabetes have neuropathy. It is generally assumed that the large variations in circulating glucose evidenced in type I diabetics contribute to the development of diabetic neuropathy and that transcriptional control of gene expression by glucose plays a major role in the progression of diabetic neuropathy. Although mechanisms have been identified for glucose regulation of gene expression in hepatocytes and pancreatic beta cells, virtually nothing is know about the molecular mechanism by which glucose regulates gene expression in neurons. This proposal will develop model systems and new technologies for the study of glucose regulation of neuronal gone expression. This work will involve a new collaboration between the Feldman and Uhler laboratories.
Three specific aims will serve to focus the proposed studies which will employ two neuroblastoma cells lines as well as rat dorsal root ganglion neurons in primary culture. In the first specific aim, a new high-throughput transfection procedure (Surface Transfection and Expression Protocol or STEP) will be adapted to the neuroblastoma cell lines and primary neurons. In the second specific aim, candidate genes transcriptionally regulated by glucose will be identified using DNA microarray hybridization. In the third specific aim, the regulation of promoters from these candidate genes as well as other well-characterized promoter elements by glucose will be studied using STEP transfection. The completion of the studies will not only identify critical genomic mechanisms responsible for the development of neurological complications of diabetes, but they will lay the foundation for the development of drug screening assays that will be essential for the testing of drugs to interfere with these mechanisms.
| Redmond, Tanya M; Ren, Xiaomei; Kubish, Ginger et al. (2004) Microarray transfection analysis of transcriptional regulation by cAMP-dependent protein kinase. Mol Cell Proteomics 3:770-9 |
| Leinninger, Gina M; Backus, Carey; Uhler, Michael D et al. (2004) Phosphatidylinositol 3-kinase and Akt effectors mediate insulin-like growth factor-I neuroprotection in dorsal root ganglia neurons. FASEB J 18:1544-6 |
