Variations of ganglioside biosynthetic pathways in the phenotype conversion from myofibroblasts to lipocytes in murine hepatic stellate cell line

GRX cell line represents hepatic stellate cell and can be transformed from an actively proliferation myofibroblast phenotype into a quiescent fat-storing lipocyte phenotype. Both express the same gangliosides (GM3, GM2, GM1 and GD1a), which are resolved as doublets on HPTLC. Upper/lower band ratio i...

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Published in:Molecular and cellular biochemistry 2007-09, Vol.303 (1-2), p.121-130
Main Authors: de Aguirres, Aline B, Mello, Paola A, Andrade, Claudia M. B, Breier, Ana Carolina, Margis, Rogério, Guaragna, Regina M, Borojevic, Radovan, Guma, Fátima C. R, Trindade, Vera M. T
Format: Article
Language:English
Subjects:
Adipocytes
Adipocytes - cytology
Adipocytes - metabolism
Adipocytes - physiology
Animals
Biosynthesis
Biosynthetic Pathways
Cell Line
Ceramide
Fatty acids
Fibroblasts - cytology
Fibroblasts - metabolism
Fumonisin B1
Galactose
Ganglioside synthesis
Gangliosides
Gangliosides - metabolism
Gene expression
Genotype & phenotype
Glycosphingolipid salvage pathways
Golgi apparatus
Hepatic stellate cell
Hepatocytes - cytology
Hepatocytes - metabolism
Inhibitors
Liver
Mice
mRNA
Myoblasts - cytology
Myoblasts - physiology
Palmitoyltransferase
Phenotype
Phenotypes
Proteins
Serine - metabolism
Serine C-Palmitoyltransferase - metabolism
Serine palmitoyltransferase
Sphingolipids - chemistry
Sphingolipids - metabolism
Sphingomyelins - chemistry
Sphingomyelins - metabolism
β-chloroalanine
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cited_by cdi_FETCH-LOGICAL-c441t-6c89e888f125b8417af0a99fe14d4efe572dc58c9352db1a237f0319f1158b213
cites cdi_FETCH-LOGICAL-c441t-6c89e888f125b8417af0a99fe14d4efe572dc58c9352db1a237f0319f1158b213
container_end_page 130
container_issue 1-2
container_start_page 121
container_title Molecular and cellular biochemistry
container_volume 303
creator de Aguirres, Aline B
Mello, Paola A
Andrade, Claudia M. B
Breier, Ana Carolina
Margis, Rogério
Guaragna, Regina M
Borojevic, Radovan
Guma, Fátima C. R
Trindade, Vera M. T
description GRX cell line represents hepatic stellate cell and can be transformed from an actively proliferation myofibroblast phenotype into a quiescent fat-storing lipocyte phenotype. Both express the same gangliosides (GM3, GM2, GM1 and GD1a), which are resolved as doublets on HPTLC. Upper/lower band ratio is increased in lipocyte-like cells and the upper band is composed by ceramides with long-chain fatty acids. This study evaluated the contribution of de novo synthesis, sphingosine and Golgi recycling pathways on ganglioside biosynthesis, in both phenotypes. Cells were preincubated with 5 mM β-chloroalanine (SPT: serine palmitoyltransferase inhibitor) or with 25 μM fumonisin B1 (ceramide synthase inhibitor) and then radiolabeled with [U-¹⁴C]galactose in the continued presence of inhibitors. Gangliosides were extracted, purified and analyzed by HPTLC. In myofibroblast-like cells, simple gangliosides use the de novo pathway while complex gangliosides are mainly synthesized by recycling pathways. In lipocyte-like cells, de novo pathway has a lesser contribution and this is in agreement with the lower activity of the committed enzyme of sphingolipid synthesis (SPT) detected in this phenotype. SPT mRNA has an identical expression in both phenotypes. It was also observed that gangliosides doublets from myofibroblast-like cells have the same distribution between triton soluble and insoluble fractions (upper band > lower band) while the gangliosides doublets from lipocyte-like cells show an inversion in the insoluble fraction (lower band > upper band) in comparison to soluble fraction. These results indicate that myofibroblast- and lipocyte-like cells have important differences between the glycosphingolipid biosynthetic pathways, which could contribute with the respective glycosphingolipid-enriched membrane microdomain's composition.
doi_str_mv 10.1007/s11010-007-9464-z
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Cells were preincubated with 5 mM β-chloroalanine (SPT: serine palmitoyltransferase inhibitor) or with 25 μM fumonisin B1 (ceramide synthase inhibitor) and then radiolabeled with [U-¹⁴C]galactose in the continued presence of inhibitors. Gangliosides were extracted, purified and analyzed by HPTLC. In myofibroblast-like cells, simple gangliosides use the de novo pathway while complex gangliosides are mainly synthesized by recycling pathways. In lipocyte-like cells, de novo pathway has a lesser contribution and this is in agreement with the lower activity of the committed enzyme of sphingolipid synthesis (SPT) detected in this phenotype. SPT mRNA has an identical expression in both phenotypes. 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T</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Variations of ganglioside biosynthetic pathways in the phenotype conversion from myofibroblasts to lipocytes in murine hepatic stellate cell line</atitle><jtitle>Molecular and cellular biochemistry</jtitle><addtitle>Mol Cell Biochem</addtitle><date>2007-09-01</date><risdate>2007</risdate><volume>303</volume><issue>1-2</issue><spage>121</spage><epage>130</epage><pages>121-130</pages><issn>0300-8177</issn><eissn>1573-4919</eissn><notes>http://dx.doi.org/10.1007/s11010-007-9464-z</notes><notes>ObjectType-Article-1</notes><notes>SourceType-Scholarly Journals-1</notes><notes>ObjectType-Feature-2</notes><notes>content type line 23</notes><notes>ObjectType-Article-2</notes><notes>ObjectType-Feature-1</notes><abstract>GRX cell line represents hepatic stellate cell and can be transformed from an actively proliferation myofibroblast phenotype into a quiescent fat-storing lipocyte phenotype. Both express the same gangliosides (GM3, GM2, GM1 and GD1a), which are resolved as doublets on HPTLC. Upper/lower band ratio is increased in lipocyte-like cells and the upper band is composed by ceramides with long-chain fatty acids. This study evaluated the contribution of de novo synthesis, sphingosine and Golgi recycling pathways on ganglioside biosynthesis, in both phenotypes. Cells were preincubated with 5 mM β-chloroalanine (SPT: serine palmitoyltransferase inhibitor) or with 25 μM fumonisin B1 (ceramide synthase inhibitor) and then radiolabeled with [U-¹⁴C]galactose in the continued presence of inhibitors. Gangliosides were extracted, purified and analyzed by HPTLC. In myofibroblast-like cells, simple gangliosides use the de novo pathway while complex gangliosides are mainly synthesized by recycling pathways. In lipocyte-like cells, de novo pathway has a lesser contribution and this is in agreement with the lower activity of the committed enzyme of sphingolipid synthesis (SPT) detected in this phenotype. SPT mRNA has an identical expression in both phenotypes. It was also observed that gangliosides doublets from myofibroblast-like cells have the same distribution between triton soluble and insoluble fractions (upper band &gt; lower band) while the gangliosides doublets from lipocyte-like cells show an inversion in the insoluble fraction (lower band &gt; upper band) in comparison to soluble fraction. These results indicate that myofibroblast- and lipocyte-like cells have important differences between the glycosphingolipid biosynthetic pathways, which could contribute with the respective glycosphingolipid-enriched membrane microdomain's composition.</abstract><cop>Netherlands</cop><pub>Boston : Springer US</pub><pmid>17440688</pmid><doi>10.1007/s11010-007-9464-z</doi><tpages>10</tpages></addata></record>
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identifier ISSN: 0300-8177
ispartof Molecular and cellular biochemistry, 2007-09, Vol.303 (1-2), p.121-130
issn 0300-8177
1573-4919
language eng
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subjects Adipocytes
Adipocytes - cytology
Adipocytes - metabolism
Adipocytes - physiology
Animals
Biosynthesis
Biosynthetic Pathways
Cell Line
Ceramide
Fatty acids
Fibroblasts - cytology
Fibroblasts - metabolism
Fumonisin B1
Galactose
Ganglioside synthesis
Gangliosides
Gangliosides - metabolism
Gene expression
Genotype & phenotype
Glycosphingolipid salvage pathways
Golgi apparatus
Hepatic stellate cell
Hepatocytes - cytology
Hepatocytes - metabolism
Inhibitors
Liver
Mice
mRNA
Myoblasts - cytology
Myoblasts - physiology
Palmitoyltransferase
Phenotype
Phenotypes
Proteins
Serine - metabolism
Serine C-Palmitoyltransferase - metabolism
Serine palmitoyltransferase
Sphingolipids - chemistry
Sphingolipids - metabolism
Sphingomyelins - chemistry
Sphingomyelins - metabolism
β-chloroalanine
title Variations of ganglioside biosynthetic pathways in the phenotype conversion from myofibroblasts to lipocytes in murine hepatic stellate cell line
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