A Study On Changes In MRNA Levels Of Phosphoenolpyruvate Carboxykinase And Pyruvate Carboxylase In Chicken Embryo

In this study, 7-8 chicken embryos were selected from 13 ~ 21d incubation period everyday, and selected 5 embryos randomly, then sampled the liver, kidney and skeletal muscle(n=5) respectively. According to the GenBank provided sequence using the Primer Express2.0 and Primer3 the primers of Pyruvate Carboxylase (PC), phosphoenolpyruvate carboxykinase mitochondrial type (PEPCK-M) and phosphoenolpyruvate carboxykinase cytosol type (PEPCK-C) were designed. Then isolated the total RNA from the liver, kidney and the skeletal muscle of chicken embryos, then, synthesized the cDNA by RT-PCR,and analyzed the cDNA fragments with the re-striction enzyme analysis and DNA sequencing analysis. Then analyzed the changes in mRNA express of PC mRNA, PEPCK-M mRNA and PEPCK-C mRNA with real-time quantitive PCR, in order to study the key enzyme of gluconeogenesis process within development of chicken embryo. The results showed that: the expression of PC mRNA in the odd number of days of incubation period 13d, 15d, 17d, 19d, 21 days in liver is relatively higher than the expression of that in kidney and skeletal muscle; The expression of PEPCK-C mRNA in kidney is relatively higher than the expression of that in liver and skeletal muscle; The expression of PEPCK-M mRNA in liver relatively higher than the expression of that in kidney and skeletal muscle, but the result were not significant (p >0.05). The expression of PC mRNA in liver is higher than the expression of PC mRNA in adult chicken, the result was significant (p < 0.05). In the incubation of 21d, the expression of PEPCK-C mRNA is significantly higher than the 13d and 15d of incubation (p < 0.05). In the incubation day of 15 the expression of PEPCK-M mRNA in liver is significantly higher than that of in the 21d and the adult (p < 0.05).In even-numbered days of incubation14d, 16d, 18d, 20 days, the expression of PC mRNA in liver is relatively higher than the expression of that in kidney and skeletal muscle; Also the expression of PEPCK-C mRNA in kidney is relatively higher than the expression of that in liver and skeletal muscle; The expression of PEPCK-M mRNA in liver is relatively higher than the expression of that in kidney and skeletal muscle, but there were no significant difference(p >0.05). The PC mRNA expression level in liver of the incubation period is significantly higher than that of in adult (p < 0.05). In case of kidney, the expression of PEPCK-C mRNA of 20d is significantly higher than that of in 14d (p < 0.05). Also, according to the Tukey-kramer analysis there were no significant changes of PEPCK-M mRNA expression in liver, kidney and skeletal muscle in even- numbered days of incubation.