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Abstract:

We have previously shown that during the adipose conversion of these cells the culture medium changed its viscoelastic properties due to the presence of hyaluronan and a chondroitin sulfate proteoglycan [Calvo, J.C., Rodbard, D., Katki, A., Chernick, S., and Yanagishita, M., 1991. Differentiation of 3T3-L1 preadipocytes with 3-isobutyl-1-methylxanthine and dexamethasone stimulates cell-associated and soluble chondroitin 4-sulfate proteoglycans. J. Biol. Chem. 266, 11237-11244., Calvo, J.C., Gandjbakhche, A.H., Nossal, R., Hascall, V.C., and Yanagishita, M., 1993. Rheological effects of the presence of hyaluronic acid in the extracellular media of differentiated 3T3-L1 preadipocyte cultures. Arch. Biochem. Biophys. 302, 468-475]. Here, we analyze the time course for the appearance of these molecules during drug-induced cell differentiation. The synthesis of both hyaluronan and the proteoglycan, was maximal at 48 h in the presence of isobutylmethylxanthine and dexamethasone, but while hyaluronan remained high after changing the culture medium, the proteoglycan dropped to almost basal levels after a few days. Northern analysis revealed the presence of message for a "versican-like" molecule as well as the possibility of alternative splicing. Three major bands of 9.39, 8.48, and 7.69 kb appeared in the analysis. These bands showed a dramatic increase in intensity when RNA from non-differentiated cells was compared to differentiating 3T3-L1 cells. In addition, when the time course of appearance for this message was analyzed, it perfectly correlated the metabolic labeling of the glycosaminoglycans during cell culture. The nucleotide sequencing of two exons revealed between a 100-94% homology with proteoglycan PG-M from murine endothelial cells. At least 13% of the proteoglycan was able to bind hyaluronan. Disruption of the synthesis of the proteoglycan molecule by exogenous addition of xyloside, did not prevent triglyceride accumulation but was inhibitory to preconfluent 3T3-L1 cell proliferation. Coating of plastic culture dishes with conditioned medium from differentiating 3T3-L1 cells, resulted in decreased cell adhesion. Cell adhesion was partially recovered after degradation of hyaluronan and chondroitin sulfate by enzymatic treatment. All these results indicate a possible role of these molecules in the observed changes in the viscoelastic properties of the culture medium, as well as open the field for a more thorough study of their role in 3T3-L1 cell proliferation and/or differentiation. © 2007 International Society of Matrix Biology.

Registro:

Documento: Artículo
Título:Role of versican and hyaluronan in the differentiation of 3T3-L1 cells into preadipocytes and mature adipocytes
Autor:Zizola, C.F.; Julianelli, V.; Bertolesi, G.; Yanagishita, M.; Calvo, J.C.
Filiación:Instituto de Biología y Medicina Experimental, Vuelta de Obligado 2490, 1428 Buenos Aires, Argentina
Facultad de Ciencias Exactas y Naturales, Departamento de Química Biológica, Universidad de Buenos Aires, Argentina
Faculty of Medicine, University of Calgary, Canada
Department of Biochemistry, Faculty of Dentistry, Tokyo Medical and Dental University, Japan
Palabras clave:Adipocytes; Differentiation; Hyaluronan; Proteoglycans; dexamethasone; glycosaminoglycan; hyaluronic acid; isobutylmethylxanthine; proteoglycan; RNA; triacylglycerol; versican; adipocyte; alternative RNA splicing; animal cell; article; cell adhesion; cell culture; cell differentiation; cell labeling; cell metabolism; cell proliferation; controlled study; culture medium; embryo; mouse; nonhuman; nucleotide sequence; priority journal; proadipocyte; protein binding; protein degradation; proteoglycan synthesis; reverse transcription polymerase chain reaction; sequence analysis; viscoelasticity; 1-Methyl-3-isobutylxanthine; 3T3-L1 Cells; Adipocytes; Animals; Base Sequence; Binding Sites; Cell Adhesion; Cell Differentiation; Cell Proliferation; Culture Media, Conditioned; Dexamethasone; Gene Expression; Glycosides; Hyaluronic Acid; Insulin; Kinetics; Mice; Molecular Sequence Data; Protein Binding; Proteochondroitin Sulfates; Sequence Homology, Nucleic Acid; Triglycerides; Versicans; Murinae
Año:2007
Volumen:26
Número:6
Página de inicio:419
Página de fin:430
DOI: http://dx.doi.org/10.1016/j.matbio.2007.04.002
Título revista:Matrix Biology
Título revista abreviado:Matrix Biol.
ISSN:0945053X
CODEN:MTBOE
CAS:RNA, 63231-63-0; dexamethasone, 50-02-2; hyaluronic acid, 31799-91-4, 9004-61-9, 9067-32-7; isobutylmethylxanthine, 28822-58-4; versican, 126968-45-4; 1-Methyl-3-isobutylxanthine, 28822-58-4; Cspg2 protein, mouse; Culture Media, Conditioned; Dexamethasone, 50-02-2; Glycosides; Hyaluronic Acid, 9004-61-9; Insulin, 11061-68-0; Proteochondroitin Sulfates; Triglycerides; Versicans, 126968-45-4; xylosides
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_0945053X_v26_n6_p419_Zizola

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Citas:

---------- APA ----------
Zizola, C.F., Julianelli, V., Bertolesi, G., Yanagishita, M. & Calvo, J.C. (2007) . Role of versican and hyaluronan in the differentiation of 3T3-L1 cells into preadipocytes and mature adipocytes. Matrix Biology, 26(6), 419-430.
http://dx.doi.org/10.1016/j.matbio.2007.04.002
---------- CHICAGO ----------
Zizola, C.F., Julianelli, V., Bertolesi, G., Yanagishita, M., Calvo, J.C. "Role of versican and hyaluronan in the differentiation of 3T3-L1 cells into preadipocytes and mature adipocytes" . Matrix Biology 26, no. 6 (2007) : 419-430.
http://dx.doi.org/10.1016/j.matbio.2007.04.002
---------- MLA ----------
Zizola, C.F., Julianelli, V., Bertolesi, G., Yanagishita, M., Calvo, J.C. "Role of versican and hyaluronan in the differentiation of 3T3-L1 cells into preadipocytes and mature adipocytes" . Matrix Biology, vol. 26, no. 6, 2007, pp. 419-430.
http://dx.doi.org/10.1016/j.matbio.2007.04.002
---------- VANCOUVER ----------
Zizola, C.F., Julianelli, V., Bertolesi, G., Yanagishita, M., Calvo, J.C. Role of versican and hyaluronan in the differentiation of 3T3-L1 cells into preadipocytes and mature adipocytes. Matrix Biol. 2007;26(6):419-430.
http://dx.doi.org/10.1016/j.matbio.2007.04.002