1 Characterization of Gene Expression of CCAAT/Enhancer-Binding Protein α, Glycogen Synthase and Glucose-6-Phosphatase in Genetically Modified Mouse Primary Hepatocytes

There are many studies concerning CCAAT/enhancer-binding protein α (C/EBPα) knock-out mice to examine the critical roles played by C/EBPα in metabolism. However, investigation on C/EBPα knock-in mice is still a completely new area. In this experiment, Northern blot hybridization was performed to characterize C/EBPα, glycogen synthase (GS) and glucose-6-phosphatase (G6Pase) gene expression in primary hepatocytes of C/EBPα knock-in mice. Though the difference of C/EBPα gene expression patterns between both wild-type and knock-in samples is subtle, the expression profiles of both GS and G6Pase genes are closely related to that of C/EBPα. This provides a line of evidence that trans-regulation function of C/EBPα on GS and G6Pase genes, which govern essential glucose metabolic processes, is still well-maintained in C/EBPα knock-in mice. Furthermore, it is observed that 18S rRNA is a better internal control for quantitative comparison of mRNA levels than ß-actin. The underlying clinical significance is related to prematurity, in which the glucose homeostasis is severely impaired as a result of the lack of activation of C/EBPα in development. Construction of C/EBPα knock-in mice models is one approach to ameliorate the physiological dysfunction of the premature human infant