Abstract:
Adipogenesis is stimulated in 3T3-L1 fibroblasts by a combination of insulin, dexamethasone and isobutylmethylxanthine, IBMX, (I+D+M). Two transcription factors are important for the acquisition of the adipocyte phenotype, C/EBP beta (CCAT enhancer-binding protein beta) and PPAR gamma (peroxisome proliferator-activated receptor gamma). IBMX increases cAMP content, which can activate protein kinase A (PKA) and/or EPAC (exchange protein activated by cAMP). To investigate the importance of IBMX in the differentiation mixture, we first evaluated the effect of the addition of IBMX on the increase of C/EBP beta and PPAR gamma and found an enhancement of the amount of both proteins. IBMX addition (I+D+M) or its replacement with a cAMP analogue, dibutyryl-cAMP or 8-(4-chlorophenylthio)-2-O′-methyl-cAMP (8CPT-2-Me-cAMP), the latter activates EPAC and not PKA, remarkably increased PPAR gamma mRNA. However, neither I+D nor any of the inducers alone, increased PPAR gamma mRNA to a similar extent, suggesting the importance of the presence of both IBMX and I+D. It was also found that the addition of IBMX or 8CPT-2-Me-cAMP was able to increase the content of C/EBP beta with respect to I+D. In agreement with these findings, a microarray analysis showed that the presence of either 8CPT-2-Me-cAMP or IBMX in the differentiation mixture was able to upregulate PPAR gamma and PPAR gamma-activated genes as well as other genes involved in lipid metabolism. Our results prove the involvement of IBMX-cAMP-EPAC in the regulation of adipogenic genes during differentiation of 3T3-L1 fibroblasts and therfore contributes to elucidate the role of cyclic AMP in this process. © 2014 Japanese Society of Developmental Biologists.
Registro:
Documento: |
Artículo
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Título: | Exchange protein activated by cyclic AMP is involved in the regulation of adipogenic genes during 3T3-L1 fibroblasts differentiation |
Autor: | Gabrielli, M.; Martini, C.N.; Brandani, J.N.; Iustman, L.J.R.; Romero, D.G.; Vila, M.D.C. |
Filiación: | Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Ciudad Universitaria, Pabellón 2, Buenos Aires, 1428, Argentina Department of Biochemistry, University of Mississippi Medical Center, 2500 North State Street, Jackson, MS, 39216, United States
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Palabras clave: | 3T3-L1 cells; Adipogenesis; CCAT enhancer-binding protein beta; Cyclic AMP; Peroxisome proliferator-activated receptor gamma; 3T3-L1 cells; adipogenesis; CCAT enhancer-binding protein beta; cyclic AMP; peroxisome proliferator-activated receptor gamma; 1-Methyl-3-isobutylxanthine; 3T3-L1 Cells; Adipogenesis; Analysis of Variance; Animals; Azo Compounds; Blotting, Western; CCAAT-Enhancer-Binding Protein-beta; Cell Count; Cell Differentiation; Cyclic AMP; Dexamethasone; DNA Primers; Gene Expression Regulation; Guanine Nucleotide Exchange Factors; Insulin; Mice; Microarray Analysis; Microscopy, Fluorescence; PPAR gamma |
Año: | 2014
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Volumen: | 56
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Número: | 2
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Página de inicio: | 143
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Página de fin: | 151
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DOI: |
http://dx.doi.org/10.1111/dgd.12114 |
Título revista: | Development Growth and Differentiation
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Título revista abreviado: | Dev. Growth Differ.
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ISSN: | 00121592
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CODEN: | DGDFA
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Registro: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00121592_v56_n2_p143_Gabrielli |
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Citas:
---------- APA ----------
Gabrielli, M., Martini, C.N., Brandani, J.N., Iustman, L.J.R., Romero, D.G. & Vila, M.D.C.
(2014)
. Exchange protein activated by cyclic AMP is involved in the regulation of adipogenic genes during 3T3-L1 fibroblasts differentiation. Development Growth and Differentiation, 56(2), 143-151.
http://dx.doi.org/10.1111/dgd.12114---------- CHICAGO ----------
Gabrielli, M., Martini, C.N., Brandani, J.N., Iustman, L.J.R., Romero, D.G., Vila, M.D.C.
"Exchange protein activated by cyclic AMP is involved in the regulation of adipogenic genes during 3T3-L1 fibroblasts differentiation"
. Development Growth and Differentiation 56, no. 2
(2014) : 143-151.
http://dx.doi.org/10.1111/dgd.12114---------- MLA ----------
Gabrielli, M., Martini, C.N., Brandani, J.N., Iustman, L.J.R., Romero, D.G., Vila, M.D.C.
"Exchange protein activated by cyclic AMP is involved in the regulation of adipogenic genes during 3T3-L1 fibroblasts differentiation"
. Development Growth and Differentiation, vol. 56, no. 2, 2014, pp. 143-151.
http://dx.doi.org/10.1111/dgd.12114---------- VANCOUVER ----------
Gabrielli, M., Martini, C.N., Brandani, J.N., Iustman, L.J.R., Romero, D.G., Vila, M.D.C. Exchange protein activated by cyclic AMP is involved in the regulation of adipogenic genes during 3T3-L1 fibroblasts differentiation. Dev. Growth Differ. 2014;56(2):143-151.
http://dx.doi.org/10.1111/dgd.12114