Abstract
Meat quality is closely related to adipose tissues in ducks, and adipogenesis is controlled by a complex network of transcription factors tightly acting at different stages of differentiation especially in ducks. The aim of this study was to establish the preadipocyte in vitro culture system and understand the biological characteristics of expansion of duck adipocyte tissue at the cellular and molecular level. We isolated pre-adipocytes from the subcutaneous fat of three breeds of duck and differentiated them into mature adipocytes using a mixture of insulin, rosiglitazone, dexamethasone, 3-isobutyl-1-methylxanthine, and oleic acid over 0,2, 4, 6, and 8 days. Successful differentiation was confirmed from the development of lipid droplets and their response to Oil Red O, and increasing numbers of lipid droplets were stained red over time. The expression of key marker genes, including peroxisome proliferator activated receptor γ (PPARγ), CCAAT/enhancer binding protein-α (C/EBPα), adipocyte fatty acid binding protein 4 (FABP4), and fatty acid synthetase (FAS), gradually increased during pre-adipocyte differentiation. Furthermore, it was verified by interference experiments that the knockdown of PPARγ directly reduced lipid production. Meanwhile we analyzed the role of unsaturated fatty acids in the production of poultry fat using different concentrations of oleic acid and found that lipid droplet deposition was highest when the concentration of oleic acid was 300 μM. We also compared the level of differentiated pre-adipocytes that were isolated from Jianchang ducks (fatty-meat duck), Cherry Valley ducks (lean-meat duck) and White-crested ducks (egg-producing duck). The proliferation and differentiation rate of pre-adipocytes derived from Jianchang ducks was higher than that of White-crested ducks. These results provide the foundation for further research into waterfowl adipogenesis.
Highlights
Anatine adipose tissue is directly associated with the production and meat quality characteristics
Previous studies had shown that the differentiation and adipogenesis in preadipocytes was a very complex process involving a large amount of gene expression.Understanding the molecular mechanism of adipogenesis can help to improve the quality of meat, increase the utilization rate of feed, and contribute to the feed conversion ratio (FCR)
Pre-adipocytes obtained after collagenase digestion that were adhered to the bottom of the flasks reached 100% confluency after 24 h in the basal culture medium (Fig 1A)
Summary
Anatine adipose tissue is directly associated with the production and meat quality characteristics. It is composed of a large number of clustered adipocytes. The proliferation and differentiation of pre-adipocytes leads to the formation of adipose tissue. Previous studies had shown that the differentiation and adipogenesis in preadipocytes was a very complex process involving a large amount of gene expression.Understanding the molecular mechanism of adipogenesis can help to improve the quality of meat, increase the utilization rate of feed, and contribute to the feed conversion ratio (FCR). Transcriptional factors including CCAAT/enhancer-binding protein (C/EBP) family including C/EBPα, C/EBPβ, and C/EBPδ [5,6], and peroxisome proliferator-activated receptor (PPAR) [7,8], as well as other genes, such as fatty acid-binding protein (FABP4) [9], lipoprotein-lipase (LPL), and fatty acid synthase (FAS) [10] both played important roles in the formation of fat. DLK1 (delta drosophila homolog-like 1 or delta-like 1 homologue) first discovered in the neuroblastoma could inhibit the differentiation of pre-adipocytes and was known as pre-adipocyte factor-1 (pref-1)[11]
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