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Katrin Mani - One of the best experts on this subject based on the ideXlab platform.
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Common traffic routes for imported spermine and endosomal Glypican-1-derived heparan sulfate in fibroblasts
Experimental cell research, 2018Co-Authors: Fang Cheng, Lars-Åke Fransson, Katrin ManiAbstract:Import of the polyamine spermine from the extracellular environment depends on the presence of cell surface heparan sulfate proteoglycans, such as Glypican-1. This proteoglycan is internalized by endocytosis, releases its heparan sulfate chains in endosomes by a nitric oxide-, copper- and amyloid precursor protein-dependent mechanism, then penetrates the membrane and is transported to the nucleus and then to autophagosomes. This process is spontaneous or induced by ascorbate depending on the growth-state of the cell. Here, we have explored possible connections between the heparan sulfate traffic route and spermine uptake and delivery in wild-type and Tg2576 mouse fibroblasts. Cells were examined by deconvolution immunofluorescence microscopy. The antibodies used were specific for spermine, Glypican-1-derived heparan sulfate, Rab7, nucleolin and a marker for autophagosomes. Endogenous immunostainable spermine was primarily associated with autophagosomes. When spermine synthesis was inhibited, imported spermine appeared in Rab7-positive endosomes. When ascorbate was added, heparan sulfate and spermine were transported to the nucleus where they colocalized with nucleolin. Spermine also appeared in autophagosomes. In a pulse-chase experiment, heparan sulfate and spermine were first arrested in late endosomes by actinomycin D treatment. During the chase, when arrest was abolished, heparan sulfate and spermine were both transported to the nucleus and targeted nucleolin. In amyloid precursor protein-/--fibroblasts, ascorbate failed to induce release of heparan sulfate and spermine remained in the endosomes. We propose that cell surface Glypican-1 carries spermine to the endosomes and that the released heparan sulfate carries spermine across the membrane into the cytosol and then to the nucleus.
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Structural Aspects of N-Glycosylations and the C-terminal Region in Human Glypican-1.
The Journal of biological chemistry, 2015Co-Authors: Wael Awad, Katrin Mani, Barbara Adamczyk, Jessica Örnros, Niclas G. Karlsson, Derek T. LoganAbstract:Glypicans are multifunctional cell surface proteoglycans involved in several important cellular signaling pathways. Glypican-1 (Gpc1) is the predominant heparan sulfate proteoglycan in the developing and adult human brain. The two N-linked glycans and the C-terminal domain that attach the core protein to the cell membrane are not resolved in the Gpc1 crystal structure. Therefore, we have studied Gpc1 using crystallography, small angle x-ray scattering, and chromatographic approaches to elucidate the composition, structure, and function of the N-glycans and the C terminus and also the topology of Gpc1 with respect to the membrane. The C terminus is shown to be highly flexible in solution, but it orients the core protein transverse to the membrane, directing a surface evolutionarily conserved in Gpc1 orthologs toward the membrane, where it may interact with signaling molecules and/or membrane receptors on the cell surface, or even the enzymes involved in heparan sulfate substitution in the Golgi apparatus. Furthermore, the N-glycans are shown to extend the protein stability and lifetime by protection against proteolysis and aggregation.
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Improving the diffraction of Glypican-1 crystals by controlled dehydration
Acta Crystallographica Section A Foundations and Advances, 2014Co-Authors: Wael Awad, Katrin Mani, Gabriel Svensson Birkedal, Marjolein M.g.m. Thunnissen, Derek T. LoganAbstract:Glypicans are heparan sulfate proteoglycans that are attached to the cell membrane surface by glycosylphosphatidylinositol anchorage. Glypican-1 (Gpc-1) is the predominant heparan sulphate proteoglycan in the developing and adult human brain and is involved in developmental morphogenesis, growth factor and cytokine signalling. We determined the crystal structure of N-glycosylated human Glypican-1 core protein at 2.55 Å resolution, which revealed a cylindrical, all α-helical fold (dimensions 120 x 30 x 30 Å), decorated with three major loops and containing the 14 cysteine residues conserved in all members of the Glypican family (1). The Gpc-1 crystals were delicate, highly fragile plates, which displayed poor isomorphism, with cell dimensions varying between different crystals. These crystals also diffracted anisotropically, reflected in a Wilson B factor that was twice as large in the c* direction as in the a* and b* directions, which limited the effective resolution to 2.9 Å in the c* direction. Recently we have shown Gpc-1 crystals to be a successful case for improvement in diffraction properties by controlled crystal dehydration using the humidity control device (HC1b), which delivers a humidified air stream of a precise relative humidity that can be used to alter the solvent content inside the crystals (2). The optimal dehydration protocol was developed by investigation of the parameters: final relative humidity RHf, dehydration rate and total incubation time Tinc. Of these, the most important was shown to be Tinc. After dehydration using the optimal protocol, the diffraction quality of the Gpc-1 crystals was clearly improved, with significant reduction in the anisotropy. This generated better, less noisy electron density maps, which allowed the building of previously disordered parts of the model and displayed well-defined side chains.
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Improvements in the order, isotropy and electron density of Glypican-1 crystals by controlled dehydration.
Acta Crystallographica Section D Biological Crystallography, 2013Co-Authors: Wael Awad, Katrin Mani, Gabriel Svensson Birkedal, Marjolein M.g.m. Thunnissen, Derek T. LoganAbstract:The use of controlled dehydration for improvement of protein crystal diffraction quality is increasing in popularity, although there are still relatively few documented examples of success. A study has been carried out to establish whether controlled dehydration could be used to improve the anisotropy of crystals of the core protein of the human proteoglycan Glypican-1. Crystals were subjected to controlled dehydration using the HC1 device. The optimal protocol for dehydration was developed by careful investigation of the following parameters: dehydration rate, final relative humidity and total incubation time Tinc. Of these, the most important was shown to be Tinc. After dehydration using the optimal protocol the crystals showed significantly reduced anisotropy and improved electron density, allowing the building of previously disordered parts of the structure.
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crystal structure of n glycosylated human Glypican 1 core protein structure of two loops evolutionarily conserved in vertebrate Glypican 1
Journal of Biological Chemistry, 2012Co-Authors: Gabriel Svensson, Wael Awad, M Hakansson, Katrin Mani, D T LoganAbstract:Glypicans are a family of cell-surface proteoglycans that regulate Wnt, hedgehog, bone morphogenetic protein, and fibroblast growth factor signaling. Loss-of-function mutations in Glypican core proteins and in glycosaminoglycan-synthesizing enzymes have revealed that Glypican core proteins and their glycosaminoglycan chains are important in shaping animal development. Glypican core proteins consist of a stable α-helical domain containing 14 conserved Cys residues followed by a glycosaminoglycan attachment domain that becomes exclusively substituted with heparan sulfate (HS) and presumably adopts a random coil conformation. Removal of the α-helical domain results in almost exclusive addition of the glycosaminoglycan chondroitin sulfate, suggesting that factors in the α-helical domain promote assembly of HS. Glypican-1 is involved in brain development and is one of six members of the vertebrate family of Glypicans. We expressed and crystallized N-glycosylated human Glypican-1 lacking HS and N-glycosylated Glypican-1 lacking the HS attachment domain. The crystal structure of Glypican-1 was solved using crystals of selenomethionine-labeled Glypican-1 core protein lacking the HS domain. No additional electron density was observed for crystals of Glypican-1 containing the HS attachment domain, and CD spectra of the two protein species were highly similar. The crystal structure of N-glycosylated human Glypican-1 core protein at 2.5 Å, the first crystal structure of a vertebrate Glypican, reveals the complete disulfide bond arrangement of the conserved Cys residues, and it also extends the structural knowledge of Glypicans for one α-helix and two long loops. Importantly, the loops are evolutionarily conserved in vertebrate Glypican-1, and one of them is involved in glycosaminoglycan class determination.
Douglas C Mcfarland - One of the best experts on this subject based on the ideXlab platform.
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the effect of syndecan 4 and Glypican 1 expression on age related changes in myogenic satellite cell proliferation differentiation and fibroblast growth factor 2 responsiveness
Comparative Biochemistry and Physiology A-molecular & Integrative Physiology, 2013Co-Authors: Laura B Harthan, Douglas C Mcfarland, S G VellemanAbstract:Abstract Satellite cells are multipotential stem cells responsible for muscle growth and regeneration. Satellite cell proliferation, differentiation, and responsiveness to fibroblast growth factor 2 (FGF2) is, in part, regulated by the heparan sulfate proteoglycans syndecan-4 and Glypican-1. Syndecan-4 and Glypican-1 expression declines with satellite cell age and may be associated with decreased satellite cell activity. The objective of the current study was to determine if overexpression of syndecan-4 and Glypican-1 would increase proliferation, differentiation and FGF2 responsiveness in satellite cells isolated from pectoralis major muscle from 16-wk-old turkeys. Overexpression of syndecan-4 and Glypican-1 did not have a significant effect on proliferation and differentiation in 1 d, 7 wk, and 16 wk satellite cells, and did not affect FGF2 responsiveness during proliferation. Expression of syndecan-4 and Glypican-1 increased differentiation at 48 h in 1 d, 7 wk, and 16 wk cells treated with FGF2. Expression of myogenic regulatory factors MyoD, myogenin, and MRF4 was affected by the overexpression of syndecan-4 and Glypican-1. However, changes in myogenic regulatory factor expression did not have a significant effect on proliferation or differentiation. These data demonstrate that syndecan-4 and Glypican-1 are likely not directly associated with the age related decrease in satellite cell activity.
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Changes in proliferation, differentiation, fibroblast growth factor 2 responsiveness and expression of syndecan‐4 and Glypican‐1 with turkey satellite cell age
Development growth & differentiation, 2013Co-Authors: Laura B Harthan, Douglas C Mcfarland, Sandra G. VellemanAbstract:Myogenic satellite cells are heterogeneous multipotential stem cells that are required for muscle repair, maintenance, and growth. The membrane-associated heparan sulfate proteoglycans syndecan-4 and Glypican-1 differentially regulate satellite cell proliferation, differentiation, fibroblast growth factor 2 (FGF2) signal transduction, and expression of the myogenic regulatory factors MyoD and myogenin. The objective of the current study was to determine the effect of age on syndecan-4 and Glypican-1 satellite cell populations, proliferation, differentiation, FGF2 responsiveness, and expression of syndecan-4, Glypican-1, MyoD, and myogenin using satellite cells isolated from the pectoralis major muscle of 1-day-old, 7-week-old and 16-week-old turkeys. Proliferation was significantly reduced in the 16-week-old satellite cells, while differentiation was decreased in the 7-week-old and the 16-week-old cells beginning at 48 h of differentiation. Fibroblast growth factor 2 responsiveness was highest in the 1-day-old and 7-week-old cells during proliferation; during differentiation there was an age-dependent response to FGF2. Syndecan-4 and Glypican-1 satellite cell populations decreased with age, but syndecan-4 and Glypican-1 were differentially expressed with age during proliferation and differentiation. MyoD and myogenin mRNA expression was significantly decreased in 16-week-old cells compared to the 1-day-old and 7-week-old cells. MyoD and myogenin protein expression was higher during proliferation in the 16-week-old cells and decreased with differentiation. These data demonstrate an age-dependent effect on syndecan-4 and Glypican-1 satellite cell subpopulations, which may be associated with age-related changes in proliferation, differentiation, FGF2 responsiveness, and the expression of the myogenic regulatory factors MyoD and myogenin.
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changes in proliferation differentiation fibroblast growth factor 2 responsiveness and expression of syndecan 4 and Glypican 1 with turkey satellite cell age
Development Growth & Differentiation, 2013Co-Authors: Laura B Harthan, Douglas C Mcfarland, S G VellemanAbstract:Myogenic satellite cells are heterogeneous multipotential stem cells that are required for muscle repair, maintenance, and growth. The membrane-associated heparan sulfate proteoglycans syndecan-4 and Glypican-1 differentially regulate satellite cell proliferation, differentiation, fibroblast growth factor 2 (FGF2) signal transduction, and expression of the myogenic regulatory factors MyoD and myogenin. The objective of the current study was to determine the effect of age on syndecan-4 and Glypican-1 satellite cell populations, proliferation, differentiation, FGF2 responsiveness, and expression of syndecan-4, Glypican-1, MyoD, and myogenin using satellite cells isolated from the pectoralis major muscle of 1-day-old, 7-week-old and 16-week-old turkeys. Proliferation was significantly reduced in the 16-week-old satellite cells, while differentiation was decreased in the 7-week-old and the 16-week-old cells beginning at 48 h of differentiation. Fibroblast growth factor 2 responsiveness was highest in the 1-day-old and 7-week-old cells during proliferation; during differentiation there was an age-dependent response to FGF2. Syndecan-4 and Glypican-1 satellite cell populations decreased with age, but syndecan-4 and Glypican-1 were differentially expressed with age during proliferation and differentiation. MyoD and myogenin mRNA expression was significantly decreased in 16-week-old cells compared to the 1-day-old and 7-week-old cells. MyoD and myogenin protein expression was higher during proliferation in the 16-week-old cells and decreased with differentiation. These data demonstrate an age-dependent effect on syndecan-4 and Glypican-1 satellite cell subpopulations, which may be associated with age-related changes in proliferation, differentiation, FGF2 responsiveness, and the expression of the myogenic regulatory factors MyoD and myogenin.
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growth and sex effects on the expression of syndecan 4 and Glypican 1 in turkey myogenic satellite cell populations
Molecular and Cellular Biochemistry, 2013Co-Authors: Yan Song, Douglas C Mcfarland, S G VellemanAbstract:The adult skeletal muscle stem cells, satellite cells, are responsible for skeletal muscle growth and regeneration. Satellite cells represent a heterogeneous cell population that differentially express cell surface markers. The membrane-associated heparan sulfate proteoglycans, syndecan-4, and Glypican-1, are differentially expressed by satellite cells during the proliferation and differentiation stages of satellite cells. However, how the population of syndecan-4- or Glypican-1-positive satellite cells changes during proliferation and differentiation, and how sex and muscle growth potential affect the expression of these genes is unknown. Differences in the amount of satellite cells positive for syndecan-4 or Glypican-1 would affect the process of proliferation and differentiation which would impact both muscle mass accretion and the regeneration of muscle. In the current study, the percentage of satellite cells positive for syndecan-4 or Glypican-1 from male and female turkeys from a Randombred Control Line 2 and a line (F) selected for increased 16-week body weight were measured during proliferation and differentiation. Growth selection altered the population of syndecan-4- and Glypican-1-positive satellite cells and there were sex differences in the percentage of syndecan-4- and Glypican-1-positive satellite cells. This study provides new information on dynamic changes in syndecan-4- and Glypican-1-positive satellite cells showing that they are differentially expressed during myogenesis and growth selection and sex affects their expression.
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Effects of 17β-estradiol on turkey myogenic satellite cell proliferation, differentiation, and expression of Glypican-1, MyoD and myogenin.
Comparative biochemistry and physiology. Part A Molecular & integrative physiology, 2013Co-Authors: Douglas C Mcfarland, C. S. Coy, Jane E. Pesall, Sandra G. VellemanAbstract:Abstract The hypothesis of this study was that 17β-estradiol (estradiol) stimulates turkey skeletal muscle growth by influencing myogenic satellite cell proliferation, differentiation, and the gene expression of selected proteins important in regulating growth and development. Increasing levels of estradiol were administered in basal medium containing additional nutrients. Female-derived pectoralis major (PM) satellite cell proliferation was stimulated by estradiol at a level of 10 − 9 M following 4 days of treatment. Male PM and biceps femoris (BF) satellite cell proliferation was increased at 10 − 12 M estradiol. Turkey embryonic myoblast proliferation, however, decreased with 10 − 9 M and 10 − 5 M estradiol following 3 days under these conditions. Estradiol had no effect on the differentiation of any of the 4 groups of cells. Likewise, Glypican-1 expression was unaffected by estradiol treatment. MyoD expression decreased in male PM but not BF cells. MyoD expression in female PM cells and embryonic myoblasts were also unaffected by estradiol administration. Estradiol decreased myogenin expression in male satellite cells, but had no effect on female cells. There was a slight decrease in myogenin expression in embryonic myoblasts. The results demonstrate a direct effect of estradiol on avian satellite cell proliferation independent of Glypican-1, and decreased expression of MyoD and myogenin in some myogenic cells, coinciding with increased cellular proliferation.
Lars-Åke Fransson - One of the best experts on this subject based on the ideXlab platform.
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Common traffic routes for imported spermine and endosomal Glypican-1-derived heparan sulfate in fibroblasts
Experimental cell research, 2018Co-Authors: Fang Cheng, Lars-Åke Fransson, Katrin ManiAbstract:Import of the polyamine spermine from the extracellular environment depends on the presence of cell surface heparan sulfate proteoglycans, such as Glypican-1. This proteoglycan is internalized by endocytosis, releases its heparan sulfate chains in endosomes by a nitric oxide-, copper- and amyloid precursor protein-dependent mechanism, then penetrates the membrane and is transported to the nucleus and then to autophagosomes. This process is spontaneous or induced by ascorbate depending on the growth-state of the cell. Here, we have explored possible connections between the heparan sulfate traffic route and spermine uptake and delivery in wild-type and Tg2576 mouse fibroblasts. Cells were examined by deconvolution immunofluorescence microscopy. The antibodies used were specific for spermine, Glypican-1-derived heparan sulfate, Rab7, nucleolin and a marker for autophagosomes. Endogenous immunostainable spermine was primarily associated with autophagosomes. When spermine synthesis was inhibited, imported spermine appeared in Rab7-positive endosomes. When ascorbate was added, heparan sulfate and spermine were transported to the nucleus where they colocalized with nucleolin. Spermine also appeared in autophagosomes. In a pulse-chase experiment, heparan sulfate and spermine were first arrested in late endosomes by actinomycin D treatment. During the chase, when arrest was abolished, heparan sulfate and spermine were both transported to the nucleus and targeted nucleolin. In amyloid precursor protein-/--fibroblasts, ascorbate failed to induce release of heparan sulfate and spermine remained in the endosomes. We propose that cell surface Glypican-1 carries spermine to the endosomes and that the released heparan sulfate carries spermine across the membrane into the cytosol and then to the nucleus.
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Involvement of Glypican-1 autoprocessing in scrapie infection
The European journal of neuroscience, 2008Co-Authors: Kajsa Löfgren, Lars-Åke Fransson, Fang Cheng, Katarina Bedecs, Katrin ManiAbstract:The copper-binding cellular prion protein (PrPC) and the heparan sulphate (HS)-containing proteoglycan Glypican-1 (Gpc-1) can both be attached to lipid rafts via their glycosylphosphatidylinositol anchors, and copper ions stimulate their cointernalization from the cell surface to endosomes. The prion protein controls cointernalization and delivers copper necessary for S-nitrosylation of conserved cysteines in the Gpc-1 core protein. Later, during recycling through endosomal compartments, nitric oxide can be released from the S-nitroso groups and catalyses deaminative degradation and release of the HS substituents. Here, by using confocal immunofluorescence microscopy, we show that normal PrPC and Gpc-1 colocalize inside GT1-1 cells. However, in scrapie-infected cells (ScGT1-1), Gpc-1 protein remained at the cell surface separate from the cellular prion protein. Scrapie infection stimulated Gpc-1 autoprocessing and the generated HS degradation products colocalized with intracellular aggregates of the disease-related scrapie prion protein isoform (PrPSc). Coimmunoprecipitation experiments demonstrated an association between Gpc-1 and PrPC in uninfected cells, and between HS degradation products and PrPSc in infected cells. Silencing of Gpc-1 expression or prevention of Gpc-1 autoprocessing elevated the levels of intracellular PrPSc aggregates in infected cells. These results suggest a role for Gpc-1 autoprocessing in the clearance of PrPSc from infected cells.
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Novel aspects of vitamin C: how important is Glypican-1 recycling?
Trends in molecular medicine, 2007Co-Authors: Lars-Åke Fransson, Katrin ManiAbstract:The reduced form of vitamin C, ascorbic acid, is well known for its function as an antioxidant and as a protective agent against scurvy. However, many recent studies indicate other functions for vitamin C in mammalian cells. Novel findings provide possible explanations for observed beneficial effects of a high intake of vitamin C on cell growth, gene transcription, host resistance to infection, uptake of polyamines and clearance of misfolded proteins. Vitamin C exerts its effects indirectly via hypoxia-inducible factor, nitric oxide synthase and the heparan sulfate proteoglycan Glypican-1, which is deglycanated in a vitamin C- and copper-dependent reaction.
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The Amyloid Precursor Protein (APP) of Alzheimer Disease and Its Paralog, APLP2, Modulate the Cu/Zn-Nitric Oxide-catalyzed Degradation of Glypican-1 Heparan Sulfate in Vivo
The Journal of biological chemistry, 2005Co-Authors: Roberto Cappai, Fang Cheng, Lars-Åke Fransson, Giuseppe D. Ciccotosto, B. Elise Needham, Colin L. Masters, Gerd Multhaup, Katrin ManiAbstract:Processing of the recycling proteoglycan Glypican-1 involves the release of its heparan sulfate chains by copper ion- and nitric oxide-catalyzed ascorbate-triggered autodegradation. The Alzheimer disease amyloid precursor protein (APP) and its paralogue, the amyloid precursor-like protein 2 (APLP2), contain copper ion-, zinc ion-, and heparan sulfate-binding domains. We have investigated the possibility that APP and APLP2 regulate Glypican-1 processing during endocytosis and recycling. By using cell-free biochemical experiments, confocal laser immunofluorescence microscopy, and flow cytometry of tissues and cells from wild-type and knock-out mice, we find that (a) APP and Glypican-1 colocalize in perinuclear compartments of neuroblastoma cells, (b) ascorbate-triggered nitric oxidecatalyzed Glypican-1 autodegradation is zinc ion-dependent in the same cells, (c) in cell-free experiments, APP but not APLP2 stimulates Glypican-1 autodegradation in the presence of both Cu(II) and Zn(II) ions, whereas the Cu(I) form of APP and the Cu(II) and Cu(I) forms of APLP2 inhibit autodegradation, (d) in primary cortical neurons from APP or APLP2 knock-out mice, there is an increased nitric oxide-catalyzed degradation of heparan sulfate compared with brain tissue and neurons from wild-type mice, and (e) in growth-quiescent fibroblasts from APLP2 knock-out mice, but not from APP knock-out mice, there is also an increased heparan sulfate degradation. We propose that the rate of autoprocessing of Glypican-1 is modulated by APP and APLP2 in neurons and by APLP2 in fibroblasts. These observation identify a functional relationship between the heparan sulfate and copper ion binding activities of APP/APLP2 in their modulation of the nitroxyl anion-catalyzed heparan sulfate degradation in Glypican-1.
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involvement of glycosylphosphatidylinositol linked ceruloplasmin in the copper zinc nitric oxide dependent degradation of Glypican 1 heparan sulfate in rat c6 glioma cells
Journal of Biological Chemistry, 2004Co-Authors: Katrin Mani, Fang Cheng, Birgitta Havsmark, Samuel David, Lars-Åke FranssonAbstract:The core protein of Glypican-1, a glycosylphosphatidylinositol-linked heparan sulfate proteoglycan, can bind Cu(II) or Zn(II) ions and undergo S-nitrosylation in the presence of nitric oxide. Cu(II)-to-Cu(I)-reduction supports extensive and permanent nitrosothiol formation, whereas Zn(II) ions appear to support a more limited, possibly transient one. Ascorbate induces release of nitric oxide, which catalyzes deaminative degradation of the heparan sulfate chains on the same core protein. Although free Zn(II) ions support a more limited degradation, Cu(II) ions support a more extensive self-pruning process. Here, we have investigated processing of Glypican-1 in rat C6 glioma cells and the possible participation of the copper-containing glycosylphosphatidylinositol-linked splice variant of ceruloplasmin in nitrosothiol formation. Confocal microscopy demonstrated colocalization of Glypican-1 and ceruloplasmin in endosomal compartments. Ascorbate induced extensive, Zn(II)-supported heparan sulfate degradation, which could be demonstrated using a specific zinc probe. RNA interference silencing of ceruloplasmin expression reduced the extent of Zn(II)-supported degradation. In cell-free experiments, the presence of free Zn(II) ions prevented free Cu(II) ion from binding to Glypican-1 and precluded extensive heparan sulfate autodegradation. However, in the presence of Cu(II)-loaded ceruloplasmin, heparan sulfate in Zn(II)-loaded Glypican-1 underwent extensive, ascorbate-induced degradation. We propose that the Cu(II)-to-Cu(I)-reduction that is required for S-nitrosylation of Glypican-1 can take place on ceruloplasmin and thereby ensure extensive Glypican-1 processing in the presence of free Zn(II) ions.
S G Velleman - One of the best experts on this subject based on the ideXlab platform.
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the effect of nutrient restriction and syndecan 4 or Glypican 1 knockdown on the differentiation of turkey pectoralis major satellite cells differing in age and growth selection
Poultry Science, 2019Co-Authors: S G Velleman, Daniel L Clark, Jeffrey R TonnigesAbstract:ABSTRACT Skeletal muscle growth is mediated by the proliferation and differentiation of satellite cells, whose activity is affected by both nutrition and the expression of syndecan-4 and Glypican-1. Previous research has not addressed if there is an interactive effect of nutrition with the expression of syndecan-4 and Glypican-1. Thus, the objective of the current study was to determine if the response of satellite cells to nutrient restriction was altered by syndecan-4 or Glypican-1 knockdown and if age and growth selection are factors. Satellite cells were isolated from pectoralis major muscle of 1-day, 7-wk, and 16-wk-old turkeys selected for increased 16-wk body weight (F line) and the randombred control (RBC2) line from which the F line was selected. Syndecan-4 or Glypican-1 expression was knocked down in both lines using small interfering RNAs along with nutrient restriction of 0 or 20% of the standard cell culture medium either applied during proliferation with subsequent normal differentiation medium (RN) or during differentiation with preceding normal proliferation medium (NR). For both lines, nutrient restriction and syndecan-4 or Glypican-1 knockdown had an independent and additive effect on satellite cell differentiation at 72h of differentiation except for 1 d satellite cells. The 1 d satellite cell differentiation was increased by RN treatment, but when combined with syndecan-4 or Glypican-1 knockdown, the increase in differentiation was negated. At 48h of differentiation, syndecan-4 knockdown in 7 and 16 wk satellite cells and Glypican-1 knockdown in 7 wk cells cancelled the effect of the RN treatment, but enhanced the effect of NR treatment at 24h of differentiation. Growth selection had little effect on the interaction between nutrient restriction and syndecan-4 or Glypican-1 knockdown. Taken together, these data demonstrate that the satellite cell response to nutrition is dependent on the expression of syndecan-4 and Glypican-1 in an age-dependent manner with growth selection having little impact.
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the effect of syndecan 4 and Glypican 1 knockdown on the proliferation and differentiation of turkey satellite cells differing in age and growth rates
Comparative Biochemistry and Physiology A-molecular & Integrative Physiology, 2018Co-Authors: S G Velleman, Daniel L Clark, Jeffrey R TonnigesAbstract:Abstract Posthatch skeletal muscle growth requires myogenic satellite cells and the dynamic expression of cell membrane-associated proteins. The membrane associated heparan sulfate proteoglycans, syndecan-4 and Glypican-1, link the satellite cell niche to the intracellular environment. Sydnecan-4 and Glypican-1 are differentially expressed with age in turkey satellite cells and their over-expression impacts both satellite cell proliferation and differentiation, but their effect on satellite cells from lines with different growth potentials is not known. The objective of the current study was to determine if syndecan-4 and Glypican-1 regulation of satellite cell proliferation and differentiation is affected by age and growth selection. Pectoralis major satellite cells isolated at 1 d, 7 and 16-wk of age from a Randombred Control 2 (RBC2) line and a 16-wk body weight (F) line selected from the RBC2 line turkeys were studied. Syndecan-4 and Glypican-1 expression was knocked down in both lines. The F-line cells proliferated faster than RBC2 line cells regardless of age, while differentiation tended to be greater in RBC2 line cells than F-line cells at each age. Syndecan-4 knockdown decreased proliferation at 7- and 16-wk but not 1 d cells, and increased differentiation at 1 d and 7 wk but not 16 wk cells. Glypican-1 knockdown differentially affected proliferation depending on cell age, whereas differentiation was decreased for 7- and 16-wk but not 1 d cells. These data suggest syndecan-4 and Glypican-1 differentially affected satellite cell function in an age-dependent manner, but had little impact on differences in proliferation and differentiation due to growth selection.
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development and growth of the avian pectoralis major breast muscle function of syndecan 4 and Glypican 1 in adult myoblast proliferation and differentiation
Frontiers in Physiology, 2017Co-Authors: S G Velleman, Yan SongAbstract:Muscle fiber number is determined around the time hatch with continued posthatch muscle growth being mediated by the adult myoblast, satellite cell, population of cells. Satellite cells are dynamic in their expression of proteins including the cell membrane associated proteoglycans, syndecan-4 and Glypican-1. These proteoglycans play roles in organizing the extracellular environment in the satellite cell niche, cytoskeletal structure, cell-to-cell adhesion, satellite cell migration, and signal transduction. This review article focuses on syndecan-4 and Glypican-1 as both are capable of regulating satellite cell responsiveness to fibroblast growth factor 2. Fibroblast growth factor 2 is a potent stimulator of muscle cell proliferation and a strong inhibitor of differentiation. Proteoglycans are composed of a central core protein defined functional domains, and covalently attached glycosaminoglycans and N-glycosylation chains. The functional association of these components with satellite cell function is discussed as well as an emerging role for microRNA regulation of syndecan-4 and Glypican-1.
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the effect of syndecan 4 and Glypican 1 expression on age related changes in myogenic satellite cell proliferation differentiation and fibroblast growth factor 2 responsiveness
Comparative Biochemistry and Physiology A-molecular & Integrative Physiology, 2013Co-Authors: Laura B Harthan, Douglas C Mcfarland, S G VellemanAbstract:Abstract Satellite cells are multipotential stem cells responsible for muscle growth and regeneration. Satellite cell proliferation, differentiation, and responsiveness to fibroblast growth factor 2 (FGF2) is, in part, regulated by the heparan sulfate proteoglycans syndecan-4 and Glypican-1. Syndecan-4 and Glypican-1 expression declines with satellite cell age and may be associated with decreased satellite cell activity. The objective of the current study was to determine if overexpression of syndecan-4 and Glypican-1 would increase proliferation, differentiation and FGF2 responsiveness in satellite cells isolated from pectoralis major muscle from 16-wk-old turkeys. Overexpression of syndecan-4 and Glypican-1 did not have a significant effect on proliferation and differentiation in 1 d, 7 wk, and 16 wk satellite cells, and did not affect FGF2 responsiveness during proliferation. Expression of syndecan-4 and Glypican-1 increased differentiation at 48 h in 1 d, 7 wk, and 16 wk cells treated with FGF2. Expression of myogenic regulatory factors MyoD, myogenin, and MRF4 was affected by the overexpression of syndecan-4 and Glypican-1. However, changes in myogenic regulatory factor expression did not have a significant effect on proliferation or differentiation. These data demonstrate that syndecan-4 and Glypican-1 are likely not directly associated with the age related decrease in satellite cell activity.
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changes in proliferation differentiation fibroblast growth factor 2 responsiveness and expression of syndecan 4 and Glypican 1 with turkey satellite cell age
Development Growth & Differentiation, 2013Co-Authors: Laura B Harthan, Douglas C Mcfarland, S G VellemanAbstract:Myogenic satellite cells are heterogeneous multipotential stem cells that are required for muscle repair, maintenance, and growth. The membrane-associated heparan sulfate proteoglycans syndecan-4 and Glypican-1 differentially regulate satellite cell proliferation, differentiation, fibroblast growth factor 2 (FGF2) signal transduction, and expression of the myogenic regulatory factors MyoD and myogenin. The objective of the current study was to determine the effect of age on syndecan-4 and Glypican-1 satellite cell populations, proliferation, differentiation, FGF2 responsiveness, and expression of syndecan-4, Glypican-1, MyoD, and myogenin using satellite cells isolated from the pectoralis major muscle of 1-day-old, 7-week-old and 16-week-old turkeys. Proliferation was significantly reduced in the 16-week-old satellite cells, while differentiation was decreased in the 7-week-old and the 16-week-old cells beginning at 48 h of differentiation. Fibroblast growth factor 2 responsiveness was highest in the 1-day-old and 7-week-old cells during proliferation; during differentiation there was an age-dependent response to FGF2. Syndecan-4 and Glypican-1 satellite cell populations decreased with age, but syndecan-4 and Glypican-1 were differentially expressed with age during proliferation and differentiation. MyoD and myogenin mRNA expression was significantly decreased in 16-week-old cells compared to the 1-day-old and 7-week-old cells. MyoD and myogenin protein expression was higher during proliferation in the 16-week-old cells and decreased with differentiation. These data demonstrate an age-dependent effect on syndecan-4 and Glypican-1 satellite cell subpopulations, which may be associated with age-related changes in proliferation, differentiation, FGF2 responsiveness, and the expression of the myogenic regulatory factors MyoD and myogenin.
Sandra G. Velleman - One of the best experts on this subject based on the ideXlab platform.
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Changes in proliferation, differentiation, fibroblast growth factor 2 responsiveness and expression of syndecan‐4 and Glypican‐1 with turkey satellite cell age
Development growth & differentiation, 2013Co-Authors: Laura B Harthan, Douglas C Mcfarland, Sandra G. VellemanAbstract:Myogenic satellite cells are heterogeneous multipotential stem cells that are required for muscle repair, maintenance, and growth. The membrane-associated heparan sulfate proteoglycans syndecan-4 and Glypican-1 differentially regulate satellite cell proliferation, differentiation, fibroblast growth factor 2 (FGF2) signal transduction, and expression of the myogenic regulatory factors MyoD and myogenin. The objective of the current study was to determine the effect of age on syndecan-4 and Glypican-1 satellite cell populations, proliferation, differentiation, FGF2 responsiveness, and expression of syndecan-4, Glypican-1, MyoD, and myogenin using satellite cells isolated from the pectoralis major muscle of 1-day-old, 7-week-old and 16-week-old turkeys. Proliferation was significantly reduced in the 16-week-old satellite cells, while differentiation was decreased in the 7-week-old and the 16-week-old cells beginning at 48 h of differentiation. Fibroblast growth factor 2 responsiveness was highest in the 1-day-old and 7-week-old cells during proliferation; during differentiation there was an age-dependent response to FGF2. Syndecan-4 and Glypican-1 satellite cell populations decreased with age, but syndecan-4 and Glypican-1 were differentially expressed with age during proliferation and differentiation. MyoD and myogenin mRNA expression was significantly decreased in 16-week-old cells compared to the 1-day-old and 7-week-old cells. MyoD and myogenin protein expression was higher during proliferation in the 16-week-old cells and decreased with differentiation. These data demonstrate an age-dependent effect on syndecan-4 and Glypican-1 satellite cell subpopulations, which may be associated with age-related changes in proliferation, differentiation, FGF2 responsiveness, and the expression of the myogenic regulatory factors MyoD and myogenin.
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Effects of 17β-estradiol on turkey myogenic satellite cell proliferation, differentiation, and expression of Glypican-1, MyoD and myogenin.
Comparative biochemistry and physiology. Part A Molecular & integrative physiology, 2013Co-Authors: Douglas C Mcfarland, C. S. Coy, Jane E. Pesall, Sandra G. VellemanAbstract:Abstract The hypothesis of this study was that 17β-estradiol (estradiol) stimulates turkey skeletal muscle growth by influencing myogenic satellite cell proliferation, differentiation, and the gene expression of selected proteins important in regulating growth and development. Increasing levels of estradiol were administered in basal medium containing additional nutrients. Female-derived pectoralis major (PM) satellite cell proliferation was stimulated by estradiol at a level of 10 − 9 M following 4 days of treatment. Male PM and biceps femoris (BF) satellite cell proliferation was increased at 10 − 12 M estradiol. Turkey embryonic myoblast proliferation, however, decreased with 10 − 9 M and 10 − 5 M estradiol following 3 days under these conditions. Estradiol had no effect on the differentiation of any of the 4 groups of cells. Likewise, Glypican-1 expression was unaffected by estradiol treatment. MyoD expression decreased in male PM but not BF cells. MyoD expression in female PM cells and embryonic myoblasts were also unaffected by estradiol administration. Estradiol decreased myogenin expression in male satellite cells, but had no effect on female cells. There was a slight decrease in myogenin expression in embryonic myoblasts. The results demonstrate a direct effect of estradiol on avian satellite cell proliferation independent of Glypican-1, and decreased expression of MyoD and myogenin in some myogenic cells, coinciding with increased cellular proliferation.
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Modulation of turkey myogenic satellite cell differentiation through the shedding of Glypican-1.
Comparative biochemistry and physiology. Part A Molecular & integrative physiology, 2012Co-Authors: Sandra G. Velleman, Yan Song, Jonghyun Shin, Douglas C McfarlandAbstract:Glypican-1 is a cell membrane heparan sulfate proteoglycan. It is composed of a core protein with covalently attached glycosaminoglycan, and N-linked glycosylated (N-glycosylated) chains, and is attached to the cell membrane by a glycosylphosphatidylinositol (GPI) linkage. Glypican-1 plays a key role in the growth and development of muscle by regulating fibroblast growth factor 2 (FGF2). The GPI anchor of Glypican-1 can be cleaved, resulting in Glypican-1 being secreted or shed into the extracellular matrix environment. The objective of the current study was to investigate the role of Glypican-1 shedding and the glycosaminoglycan and N-glycosylated chains in regulating the differentiation of turkey myogenic satellite cells. A Glypican-1 construct without the GPI anchor was cloned into the mammalian expression vector pCMS-EGFP, and Glypican-1 without the GPI anchor and glycosaminoglycan and N-glycosylated chains were also cloned. These constructs were co-transfected into turkey myogenic satellite cells with a small interference RNA targeting the GPI anchor of endogenous Glypican-1. The soluble Glypican-1 mutants were not detected in the satellite cells but in the cell medium, suggesting the secretion of the soluble Glypican-1 mutants. Soluble Glypican-1 increased satellite cell differentiation and enhanced myotube formation in the presence of exogenous FGF2. The increase in differentiation was supported by the elevated expression of myogenin. In conclusion, the shedding of Glypican-1 from the satellite cell surface acts as a positive regulator of satellite cell differentiation and sequesters FGF2, permitting further differentiation.
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Effect of Glypican-1 covalently attached chains on turkey myogenic satellite cell proliferation, differentiation, and fibroblast growth factor 2 responsiveness
Poultry science, 2010Co-Authors: Yan Song, Douglas C Mcfarland, Karl E. Nestor, Sandra G. VellemanAbstract:ABSTRACT Glypican-1 is a cell membrane heparan sulfate proteoglycan that is composed of a core protein and covalently attached glycosaminoglycan (GAG) chains and N-linked glycosylated (N-glycosylated) chains. The Glypican-1 GAG chains are required for cell differentiation and responsiveness to fibroblast growth factor 2 (FGF2). The role of Glypican-1 N-glycosylated chains in regulating cell activities has not been reported. The objective of the current study was to investigate the role of Glypican-1 N-glycosylated chains and the interaction between N-glycosylated and GAG chains in turkey myogenic satellite cell proliferation, differentiation, and FGF2 responsiveness. The wild-type turkey Glypican-1 and turkey Glypican-1 with mutated GAG chain attachment sites were cloned into the pCMS-EGFP mammalian expression vector and were used as templates to generate Glypican-1 N-glycosylated 1-chain and no-chain mutants with or without GAG chains by site-directed mutagenesis. The wild-type Glypican-1 and all Glypican-1 N-glycosylated 1-chain and no-chain mutants with or without GAG chains were transfected into turkey myogenic satellite cells. Cell proliferation, differentiation, and FGF2 responsiveness were measured. The overexpression of Glypican-1 N-glycosylated 1-chain and no-chain mutants without GAG chains increased cell proliferation and differentiation compared with the wild-type Glypican-1 but not the Glypican-1 N-glycosylated mutants with GAG chains attached. Cells overexpressing Glypican-1 N-glycosylated mutants with or without GAG chains increased cell responsiveness to FGF2 compared with wild-type Glypican-1. These data suggest that Glypican-1 N-glycosylated chains and GAG chains are critical in regulating turkey myogenic satellite cell proliferation, differentiation, and responsivness to FGF2.
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The Effect of Glypican-1 Glycosaminoglycan Chains on Turkey Myogenic Satellite Cell Proliferation, Differentiation, and Fibroblast Growth Factor 2 Responsiveness
Poultry science, 2007Co-Authors: X. Zhang, Douglas C Mcfarland, Caini Liu, Karl E. Nestor, Sandra G. VellemanAbstract:Abstract The Glypicans are a family of cell-surface heparan sulfate proteoglycans consisting of a core protein covalently attached with glycosaminoglycans (GAG). Only Glypican-1 is expressed in skeletal muscle and increases in expression during myoblast differentiation. Previous studies have suggested that Glypican-1 influences fibroblast growth factor 2 (FGF2) signaling pathway by its heparan sulfate chains. Fibroblast growth factor 2 is a potent stimulator of muscle cell proliferation and an intense inhibitor of differentiation. To investigate the functional contribution of each GAG chain attachment site, a turkey Glypican-1 full length cDNA (1,650 bp, Gen-Bank accession number AY551002) was cloned into the pCMS-EGFP vector and mutated at 2 or all 3 potential GAG attachment sites at Ser483, Ser485, and Ser487 to obtain 1-chain and no-chain mutants, respectively. The unmutated Glypican-1, 1-chain, and no-chain mutants, and the pCMS-EGFP vector without an insert were transfected into turkey myogenic satellite cells. The transfected cell cultures were assayed for cell proliferation, differentiation, and FGF2 responsiveness. The overexpression of Glypican-1 increased FGF2 responsiveness during proliferation compared with the 1-chain, no-chain mutants, and the pCMS-EGFP vector without an insert, but there was no significant interaction between FGF2 and Glypican-1. The overexpression of Glypican-1 also increased differentiation but did not affect proliferation when compared with the 1-chain, no-chain mutants, and the pCMS-EGFP vector without an insert. To support the overexpression data, Glypican-1 expression was reduced using a small interfering RNA against turkey Glypican-1. Inhibition of Glypican-1 expression decreased myogenic satellite cell proliferation, differentiation, and FGF2 responsiveness during proliferation. These data indicate that Glypican-1 function requires the GAG chain attachment sites for myogenic satellite cell FGF2 responsiveness during proliferation and to affect the process of differentiation.