The differentiation of preadipocytes and gene expression related to adipogenesis in ducks (Anas platyrhynchos)

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.

Adipogenic and Myogenic Potentials of Chicken Embryonic Fibroblasts in vitro: Combination of Fatty Acids and Insulin Induces Adipogenesis

“…In the current study, insulin alone resulted in neither a significant increase in lipid accumulation, nor an upregulation of genes involved in adipogenesis (Klf6, Znf423, PPARg and FABP4) (Gupta et al, 2010;Hutley et al, 2003;Li et al, 2005;Shang et al, 2014;Wang et al, 2018), fatty acid transport (Fatp4) (Hall et al, 2005), and TAG synthesis (Acsl1, Agpat1, and Dgat2) (Mashek et al, Wang et al, 2017). Insulin is one of the potent adipogenic hormones which promotes differentiation of a murine preadipocyte cell line (3 T3-L1) into adipocytes without supplementation of fatty acids (Klemm et al, 2001), by increasing de novo lipogenesis from glucose.…”

gga-miRNA-18b-3p Inhibits Intramuscular Adipocytes Differentiation in Chicken by Targeting the ACOT13 Gene

“…Recent research found that PPARs had an influence on adipocyte differentiation [42,43] and fatty acid oxidation [44,45]. The marker gene, FABP [46], has been proven to have the same function in the PPARs signaling pathway. In the pathway of fatty acid metabolism and fatty acid degradation, ACOX1 [47] is also involved in fatty acid oxidation, while FASN [48] and ELOVL5 [49] are related to fatty acid biosynthesize.…”

Dynamics of transcriptome changes during subcutaneous preadipocyte differentiation in ducks

“…Recent studies showed that the process of preadipocyte differentiation in chicken has both similarities and differences with mammals [18]. Very limited studies based on candidate genes showed that several well-known TFs have similar expression patterns in chickens [19] and ducks [20] during adipocyte differentiation.…”