Key Laboratory of Animal Genetics and Breeding and Molecular Design of Jiangsu Province, College of Animal Science and Technology, Yangzhou University, Yangzhou, Jiangsu, China.
Waterfowl Institute of Zhenjiang City, Dantu, China.
PLoS One. 2018 May 17;13(5):e0196371. doi: 10.1371/journal.pone.0196371. eCollection 2018.
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.
鸭肉品质与鸭体脂肪组织密切相关,脂肪生成受转录因子的复杂网络调控,这些转录因子在分化的不同阶段发挥紧密作用,尤其是在鸭体中。本研究旨在建立鸭前体脂肪细胞体外培养体系,从细胞和分子水平上理解鸭脂肪组织扩增的生物学特性。我们从三个鸭品种的皮下脂肪中分离前体脂肪细胞,并在 0、2、4、6 和 8 天的时间内,用胰岛素、罗格列酮、地塞米松、3-异丁基-1-甲基黄嘌呤和油酸混合物将其分化为成熟脂肪细胞。通过脂滴的形成及其对油红 O 的反应来确认分化成功,随着时间的推移,越来越多的脂滴被染成红色。关键标记基因(包括过氧化物酶体增殖物激活受体 γ(PPARγ)、CCAAT/增强子结合蛋白-α(C/EBPα)、脂肪细胞脂肪酸结合蛋白 4(FABP4)和脂肪酸合成酶(FAS))在前体脂肪细胞分化过程中逐渐增加。此外,通过干扰实验验证了 PPARγ 的敲低直接减少了脂质的产生。同时,我们用不同浓度的油酸分析了不饱和脂肪酸在禽类脂肪生产中的作用,发现当油酸浓度为 300 μM 时,脂滴沉积最高。我们还比较了从建昌鸭(高脂肪肉鸭)、樱桃谷鸭(低脂肪肉鸭)和白绒鸭(产蛋鸭)中分离的分化前体脂肪细胞的水平。建昌鸭前体脂肪细胞的增殖和分化率高于白绒鸭。这些结果为进一步研究水禽脂肪生成奠定了基础。