Expression And Regulatory Mechanism Of The Adipor1Gene In Different Varieties Of Duck

The AdipoRl gene is strongest affinity with globular adiponectin cytokines, expressed abundantly in skeletal muscle, mainly mediated by globular adiponectin involved in carbohydrate and lipid metabolism, maintain homeostasis, and the accumulation of body fat was negatively correlated, with insulin resistance, anti-atherosclerosis directly or indirectly related. Therefore the strength of the adiponectin functions depends on the level of AdipoRl expression in the corresponding tissue. MicroRNAs(miRNAs) is a class derived from chromosomal non-coding region small RNA molecules, which is about20nt.And its role is to regulate expression of target genes in the post-transcriptional level, primarily inhibit the expression of target genes. MiRNAs is play an important physiological functions in the process of regulation of gene expression, signal transduction, cell proliferation and differentiation, apoptosis, the body of cancer prevention and so on. In particular, miRNAs, as an important factor of mediation metabolism in the post-transcriptional level, is involved in the body’s energy homeostasis, regulating adipocyte differentiation. Whereas AdipoRl gene expression profiles in various tissues of the duck has not been reported in detail, view of the great significance in the treatment of obesity with the AdipoRl mediated adiponectin, we have designed this experiment, which detect AdipoRl expression profiles of the organization and temporal in the different varieties ducks, then explore the role of miR-148a during the expression of duck AdipoRl. We hope the results was studied in this research to provide some reference information in the fat accumulation regulatory mechanism and duck breeding.Tissue expression pattern of AdipoRl was detected by Real-Time PCR with β-actin as internal standard in breast muscle, leg muscle, kidney and other organizations of the10-week-old Liancheng white duck, Jinling white duck, Cherry Valley duck and Putian black duck. Moreover, timing expression profiles of AdipoRl in the chest muscles, leg muscle and liver at the2th,4th,6th,8th,10th week was detected by the same way.The main results are as follows: 1. The AdipoRl gene expression levels fluctuated considerably in breast muscle at different points, but basically wavy variation in four varieties of duck. In the duck of4th and8th week, the express of AdipoRl gene was significantly higher than other points(.P<0.05), expressed in10th week were significantly or very significantly less than8th week, simultaneously, the smallest decline of Liancheng white duck expression was significantly higher than the other three varieties of duck(P<0.05).2. The AdipoRl gene was expressed consistently in four varieties of duck legs. Early expression level was very low, with the rapid growth of the leg,4th week, AdipoRl accumulation was increased rapidly and significantly higher than the other points (P<0.01), in which Cherry Valley Duck, the highest expression,was22times higher than jingling white duck,was10times higher than liangcheng white duck,late expression flatten, and continuing in a lower abundance.At2th,6th,8th,10th week, the expression levels of Jinling white duck were intermediate levels between the two parents Liancheng white duck and Cherry Valley duck,which was reflected heterosis.3. AdipoRl gene expression pattern was different in the four varieties of duck liver tissue, with a full difference. Jinling white duck and Cherry Valley duck’s expression pattern was basically the same, the lower amount of pre-expression, the upward accumulation of the latter part. Liancheng white duck and Putian black duck’s expression level fluctuated greatly. Low starting point was accumulated in Liancheng white duck liver, and the highest expression was at4th week. While high starting point was accumulated in Putian black duck liver, moreover, the highest expression level was at10th week, and was significantly higher than the other three varieties of duck(P <0.01).4. The tissue expression of AdipoRl was detected using Real-Time PCR. The results showed that the tissue expression pattern of AdipoRl mRNA was same in liver, gizzard and other8kinds of organizations of the four varieties of duck. Expression in cardiac muscle, intestine and gizzard was very little, expression in liver, proventriculus and kidney was no significant difference with leg. In particular, it was the highestly expressed in breast muscle and higher than the other organizations hundreds even thousands of times.3’UTR fragment of the AdipoRl gene was cloned by nested PCR, which was83bp.Through the optimization of the experimental conditions, it was found that:MiR-148a could promote the expression of the of AdipoRl gene in duck skeletal muscle, not only the mRNA level was significantly higher, but also the protein level was a lot of accumulation. Simultaneously, the PPARy gene is a marker of lipid metabliosm, which expression was significantly down in the duck skeletal muscle cells, indicated that the lipid metabolism was enhanced, and the fatty acid oxidation was increased in the cells. Thus, ti is suggested that miR-148a could promote AdipoR1gene expression and promote lipid metabolism in the duck skeletal muscle cells.