Development of a Novel Pck-1: eGFP Reporter Zebrafish Line for the Discovery and Evaluation of Potential Anti-Diabetic Drugs

• Main Author: Hui, Wing
• Other Authors: Wen, Xiao-Yan
• Language: en_ca
• Published: 2013
• Subjects: Diabetes Mellitus; gluconeogenesis; PEPCK; Pck-1; zebrafish; drug discovery; transgenics; reporter line; high throughput screening; transposon; gene regulation; novel drugs; drug screen; microinjection; deletional analysis; physiology; 0719
• Online Access: http://hdl.handle.net/1807/42881

Overexpression of Phosphoenolpyruvate carboxykinase - cytosolic (PEPCK, encoded by Pck-1 gene) has been found to be associated with the prevalence of hyperglycemia in Type 2 Diabetes Mellitus (T2DM) patients. The Pck-1 enzyme catalyzes the rate limiting step in endogenous glucose production. The aims of this study are to develop a Pck-1:eGFP reporter zebrafish and validate it as a potential tool for the screening of novel anti-diabetic compounds. 3.6 kb zebrafish Pck-1 promoter fragment was cloned and a Pck-1:eGFP expression vector was constructed. After DNA microinjection, we generated Pck-1:eGFP reporter zebrafish with strong eGFP expression in developing liver. Validation studies confirmed that Pck-1:eGFP zebrafish embryos responded to treatment of glucose, cAMP and dexamethasone, metformin and rosiglitazone similarly to that of humans. This novel Pck-1:eGFP reporter fish line can serve as a tool for the screening and development of novel anti-diabetic drugs that may have potential in the treatment of T2DM.