Peroxisome proliferator-activated receptor γ isoforms differentially regulate preadipocyte proliferation, apoptosis, and differentiation in chickens
| Autor: | Jiaxin Huang, Bolin Ning, Fang Mu, Yang Jing, Ning Wang, Xiaohong Yan, Yaqi Guo, Xin You, Tingting Cui, Hui Li |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Programmed cell death
preadipocyte proliferation Response element Peroxisome proliferator-activated receptor Genetics and Molecular Biology adipogenesis 03 medical and health sciences Adipocytes Animals Protein Isoforms Electrophoretic mobility shift assay lcsh:SF1-1100 030304 developmental biology Cell Proliferation chemistry.chemical_classification 0303 health sciences Reporter gene Retinoid X receptor alpha Cell growth 0402 animal and dairy science apoptosis Cell Differentiation 04 agricultural and veterinary sciences General Medicine 040201 dairy & animal science Cell biology PPAR gamma chemistry Adipogenesis Animal Science and Zoology lcsh:Animal culture Chickens PPARγ isoforms |
| Zdroj: | Poultry Science Poultry Science, Vol 99, Iss 12, Pp 6410-6421 (2020) |
| ISSN: | 1525-3171 0032-5791 |
| Popis: | Peroxisome proliferator-activated receptor γ (PPARγ) has 2 protein isoforms (PPARγ1 and PPARγ2) generated by alternative promoter usage and alternative splicing. However, their functional uniqueness and similarity remain unclear. In the study, we investigated the effects of lentivirus-mediated overexpression of PPARγ1 and PPARγ2 on proliferation, apoptosis, and differentiation of the immortalized chicken preadipocytes. Cell Counting Kit–8 assay showed PPARγ1 and PPARγ2 overexpression markedly suppressed cell proliferation, and fluorescence activated cell sorting analysis showed that PPARγ1 and PPARγ2 overexpression caused cell cycle arrest at G0/G1 phase. Cell death detection ELISA analysis showed both PPARγ1 and PPARγ2 overexpression induced cell apoptosis. Oil red O staining and gene expression analysis showed both PPARγ1 and PPARγ2 overexpression promoted preadipocyte differentiation. In the presence of PPARγ ligand, rosiglitazone, PPARγ2 overexpression was more potent in inducing apoptosis, promoting adipogenesis, and suppressing cell proliferation than PPARγ1 overexpression. We further explored the molecular basis for their functional differences. Reporter gene assay showed that under ligand conditions, PPARγ2 overexpression resulted in 1.68-fold increase in transcription activity compared with PPARγ1. Electrophoretic mobility shift assay showed both PPARγ1 and PPARγ2 could bind to PPAR response element (PPRE) as heterodimer with retinoid X receptor alpha, and by comparison, PPARγ2 had a higher affinity for PPRE than PPARγ1. Reporter gene assay showed expression PPARγ1 and PPARγ2 similarly induced fatty acid synthase and adipocyte fatty acid–binding protein promoter activity but differentially induced lipoprotein lipase and perilipin 1 promoter activities. Coimmunoprecipitation analysis showed that PPARγ1 and PPARγ2 interacted similarly with the coactivators, Tat-interacting protein 60. Taken together, our results demonstrate that PPARγ1 and PPARγ2 differentially regulate preadipocyte proliferation, apoptosis, and differentiation as a result of their distinct and overlapping molecular functions. |
| Databáze: | OpenAIRE |
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