[PPARgamma and C/EBPalpha mRNA expression in the primary culture of rat preadipocytes during differentiation].
To investigate the cell differentiation and the expression profiles of PPARy and C/EBPalpha mRNA, the rat preadipocytes were cultured after dispersed by enzymolysis of collagenase type I and the cell shape were observed under the microscope. The MTT method was adopted to determine the growth curve. The expression levels of PPARgamma and C/EBPalpha mRNA were also detected by relative quantitative RT-PCR. The growth of preadipocytes demonstrated an S-shaped or sigmoid curve. The PPARgamma mRNA in rat preadipocytes showed rather low transcript at day 3, but its expression level markedly increased by 2.5 fold at day 5 and remained at a higher level till day 9. Similarly, the expression level of C/EBPalpha mRNA in rat preadipocytes could hardly be detected at day 3 and dramatically increased at day 7. There was a 2.3 fold increase of C/EBPalpha mRNA expression level at day 7 compared with that at day 5. In a conclusion, this study suggested that both the initiating and maintaining of differentiation process were regulated by PPARgamma, while C/EBPalpha was likely to participate in the maintaining of differentiation process only.
Chen
Sheng Feng
SF
College of Animal Science, South China Agricultural University, Guangzhou 510642.
Zhu
Xiao Tong
XT
Shu
Gang
G
Gao
Shu Jing
SJ
Gao
Ping
P
Jiang
Qing Yan
QY
D017667:Adipocytes / Q000166:cytology / Q000378:metabolism*; D000818:Animals; D022763:CCAAT-Enhancer-Binding Protein-alpha / Q000235:genetics*; D002454:Cell Differentiation / Q000235:genetics*; D002478:Cells, Cultured; D020869:Gene Expression Profiling; D047495:PPAR gamma / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors
D022763:CCAAT-Enhancer-Binding Protein-alpha; D047495:PPAR gamma; D012333:RNA, Messenger