Role of versican and hyaluronan in the differentiation of 3T3-L1 cells into preadipocytes and mature adipocytes
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...
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2007
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| Acceso en línea: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_0945053X_v26_n6_p419_Zizola http://hdl.handle.net/20.500.12110/paper_0945053X_v26_n6_p419_Zizola |
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paper:paper_0945053X_v26_n6_p419_Zizola |
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dspace |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
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 |
| spellingShingle |
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 Julianelli, Vanina Laura Bertolesi, Gabriel Esteban Calvo, Juan Carlos Role of versican and hyaluronan in the differentiation of 3T3-L1 cells into preadipocytes and mature adipocytes |
| topic_facet |
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 |
| description |
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. |
| author |
Julianelli, Vanina Laura Bertolesi, Gabriel Esteban Calvo, Juan Carlos |
| author_facet |
Julianelli, Vanina Laura Bertolesi, Gabriel Esteban Calvo, Juan Carlos |
| author_sort |
Julianelli, Vanina Laura |
| title |
Role of versican and hyaluronan in the differentiation of 3T3-L1 cells into preadipocytes and mature adipocytes |
| title_short |
Role of versican and hyaluronan in the differentiation of 3T3-L1 cells into preadipocytes and mature adipocytes |
| title_full |
Role of versican and hyaluronan in the differentiation of 3T3-L1 cells into preadipocytes and mature adipocytes |
| title_fullStr |
Role of versican and hyaluronan in the differentiation of 3T3-L1 cells into preadipocytes and mature adipocytes |
| title_full_unstemmed |
Role of versican and hyaluronan in the differentiation of 3T3-L1 cells into preadipocytes and mature adipocytes |
| title_sort |
role of versican and hyaluronan in the differentiation of 3t3-l1 cells into preadipocytes and mature adipocytes |
| publishDate |
2007 |
| url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_0945053X_v26_n6_p419_Zizola http://hdl.handle.net/20.500.12110/paper_0945053X_v26_n6_p419_Zizola |
| work_keys_str_mv |
AT julianellivaninalaura roleofversicanandhyaluronaninthedifferentiationof3t3l1cellsintopreadipocytesandmatureadipocytes AT bertolesigabrielesteban roleofversicanandhyaluronaninthedifferentiationof3t3l1cellsintopreadipocytesandmatureadipocytes AT calvojuancarlos roleofversicanandhyaluronaninthedifferentiationof3t3l1cellsintopreadipocytesandmatureadipocytes |
| _version_ |
1768544233987244032 |
| spelling |
paper:paper_0945053X_v26_n6_p419_Zizola2023-06-08T15:53:50Z Role of versican and hyaluronan in the differentiation of 3T3-L1 cells into preadipocytes and mature adipocytes Julianelli, Vanina Laura Bertolesi, Gabriel Esteban Calvo, Juan Carlos 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 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. Fil:Julianelli, V. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Bertolesi, G. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Calvo, J.C. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2007 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_0945053X_v26_n6_p419_Zizola http://hdl.handle.net/20.500.12110/paper_0945053X_v26_n6_p419_Zizola |