Expression of CCAAT/Enhancer Binding Proteins during Porcine Preadipocyte Differentiation
| Autor: | Z.K. Yu, G.J. Hausman |
|---|---|
| Rok vydání: | 1998 |
| Předmět: |
Cytotoxicity
Immunologic Time Factors Stromal cell Swine Cellular differentiation Blotting Western Cell Count Biology Dexamethasone Western blot Adipocytes Image Processing Computer-Assisted medicine Animals Insulin Protein Isoforms Cells Cultured Cell Nucleus Ccaat-enhancer-binding proteins medicine.diagnostic_test Antibodies Monoclonal Nuclear Proteins Cell Differentiation Cell Biology Lipid Metabolism Immunohistochemistry Molecular biology DNA-Binding Proteins Blot Adipogenesis Cell culture Antigens Surface CCAAT-Enhancer-Binding Proteins Stromal Cells Biomarkers Fetal bovine serum |
| Zdroj: | Experimental Cell Research. 245:343-349 |
| ISSN: | 0014-4827 |
| Popis: | The expression of three CCAAT/enhancer-binding proteins (C/EBPs) was examined with immunocytochemistry and Western blot analysis during preadipocyte differentiation in porcine stromal vascular (S-V) cell cultures. Regardless of treatment and time in culture, immunoreactivity for all three C/EBP isoforms was restricted to cell nuclei. At day 1, 50 +/- 6% of S-V cells were C/EBPdelta positive, whereas 13 +/- 3 and 11.7 +/- 3% of S-V cells were AD-3 and C/EBPalpha positive, respectively. After 3 days of seeding in fetal bovine serum (FBS) and dexamethasone (DEX), C/EBPdelta; AD-3, and C/EBPalpha-positive cells increased to 67 +/- 5, 42 +/- 4, and 32 +/- 3%, respectively. Double staining clearly showed that most of the C/EBPalpha reactive cells had not accumulated appreciable lipid after 3 days of FBS + DEX. Following 3 days of insulin treatment, the percentage of C/EBPdelta cells was 50 +/- 6, whereas the percentage of AD-3- and C/EBPalpha-positive cells was 41 +/- 4 and 31 +/- 3, respectively. After insulin treatment all fat cells were AD-3, C/EBPalpha, and C/EBPdelta positive. Double staining demonstrated that fat cells were C/EBPdelta reactive throughout the culture period. Western blotting showed changes in C/EBP isoform expression that were consistent with the immunocytochemical results. We conclude that C/EBPalpha is a terminal differentiation marker which is expressed later than AD-3 but further studies are needed to determine the relationship between C/EBPdelta and adipogenesis in porcine S-V cultures. |
| Databáze: | OpenAIRE |
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