The Study Of Changes In MRNA Levels Of G6P And FBP In Chicken Embryo

To grasp expression rule of gluconeogenesis of glucose -6– phosphatase (G6P) and fructose-1,6- bisphosphatase (FBP, liver type FBP-1, and muscle type FBP-2), the content of G6P and FBP were studid in liver, kidney and muscle at chick embryo period of 13 to 21 days. According to G6P and FBP gene sequence reported in GenBank, the primers were designed with the Primer Express2.0 (Applied Biosystems Inc.) and Primer3 (http://frodo.wi.mit.edu/cgi-bin/primer3/primer3_www.cgi). 7-8 chicken embryos were selected at incubation period of 13 ~ 21 days, and 5 embryos were selected randomly. Then the samples of liver, kidney and the muscle (n=5) were taken respectively, and also the samples of liver, kidney and muscle were taken respectively from chick embryo of white Lai Hang chicken (66weeks old, Julia) (n=4). The tRNA were isolated from liver, kidney and muscle, and the cDNA were synthesized by RT-PCR. Then the cDNA were extracted by agarose gel electrophoresis, confirmed by sequence analysis and the content was decided by the real-time quantitative PCR. After treatment of statistical analysis, the results showed that the content of expressed G6P in liver and kidney were relatively higher than that in muscle at incubation period of 13, 15, 17, 19 and 21 days, but the results were not significant. In addition, the content expressed G6P at incubation period of 19 days was significantly higher than that in adult in liver (p <0.05). Also, the mRNA expression level of FBP-1 was relatively higher in the liver than that in the kidney and muscle. In the case of liver, mRNA expression level at incubation period of 13 and 15 days was significantly higher than that in adult(p < 0.05). The mRNA expression level of FBP-2 was relatively higher in the muscle than that in the kidney and muscle. And in the case of muscle, mRNA expression level of adult period was significantly higher (p < 0.05) than that of various incubation days, also, the mRNA expression level of FBP-2 at 19 days of incubation was significantly higher than that at 13 and 15 days of incubation (p <0.05). At even-numbered days of incubation period of 14,16,18 and 20 days,content of expressed G6P was significantly higher at 14 days than that at 18 days and 20 days in muscle (p <0.05). mRNA expression level of FBP-1 was relatively higher in the liver than that in the kidney and muscle, and there was no significant difference in the mRNA expression level in the liver, kidney and muscle at the 14, 16, 18 and 20 days of incubation (p >0.05). The mRNA expression level of FBP-2 was relatively higher in the muscle than that in the liver and kidney. And mRNA expression level of FBP-2 at 18 days of incubation was significantly higher than that at 14 and 16 days of incubation.