Synthesis and secretion of proteoglycans by cultured chondrocytes

Madsen, Kjell; Holmström, Sofia Björnfot; Ostrowski, K.

doi:10.1016/0014-4827(83)90170-2

Cited by 14 publications

(4 citation statements)

References 26 publications

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“…The inability of DMD cells to recover spreading ability to the same extent as normal cells may thus be a consequence of a greater delay in the reconstitution of the GERL secretory pathway compared with normal fibroblasts. The pleiotropic action of monensin could distinguish between normal and DMD cells in other ways [for example, ionic substitution in model and cell membranes (39) and microtubule organization (40)], but this does not seem probable. However, our observations on seeding efficiency after prolonged exposure to the ionophore suggest no damage to the biochemical mechanisms required for cell-substrate attachment.…”

Section: Discussionmentioning

confidence: 99%

Monensin-induced inhibition of cell spreading in normal and dystrophic human fibroblasts.

Pizzey¹,

Witkowski²,

Jones³

1984

Proc. Natl. Acad. Sci. U.S.A.

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Cultured skin fibroblasts from normal individuals and from patients with Duchenne muscular dystrophy spread equally rapidly when seeded on a glass substratum. Exposure to the ionophore monensin substantially suppresses normal and dystrophic fibroblast spreading in serum-free media for up to at least 100 min. Preincubation of normal fibroblasts with monensin causes a further reduction in cell spreading. Dystrophic fibroblasts fail to spread as well as normal cells after monensin preincubation. Such findings indicate that there is a delay in the secretion of functional adhesive surface proteins in monensin-preincubated normal fibroblasts and that this lag period is significantly longer in dystrophic fibroblasts. These data are consistent with findings of altered adhesive and secretory properties of fibroblasts from patients with Duchenne muscular dystrophy. 27) and others (28) have found that DMD fibroblasts appear to be less adhesive than normal cells, and this may be related to altered cell surface properties in DMD (29). We have studied the spreading ability of normal and DMD fibroblasts under conditions in which protein secretion is altered by exposure to monensin and report here that there is no difference in the rate or extent of radial spreading between normal and DMD fibroblasts in serum-free media. However, although monensin suppresses the spreading of normal and DMD cells to a similar degree, we demonstrate that, after prolonged monensin preincubation, DMD fibroblasts fail to spread as efficiently as normal cells, especially when monensin is removed from the seeding media. These results are considered in relation to a possible defect in the secretory and adhesive properties of cultured DMD fibroblasts.

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Section: Discussionmentioning

confidence: 99%

Monensin-induced inhibition of cell spreading in normal and dystrophic human fibroblasts.

Pizzey¹,

Witkowski²,

Jones³

1984

Proc. Natl. Acad. Sci. U.S.A.

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“…Monensin may inhibit the conversion of oligosaccharides from the high mannose type to the complex type (Ledger et al, 1983;Sai and Tanzer, 1984) and interrupt Golgi transport at a point where the oligosaccharide product has not yet acquired galac-tose, fucose, and sialic acid (Tartakoff, 1983a,b). In cultured chondrocytes, monensin causes an inhibition of sulfate incorporation into chondroitin sulfate (Mitchell and Hardingham, 1982;Nishimoto et al, 1982;Burditt et al, 1985) and blocks intracellular transport of proteoglycans (Kajiwara and Tanzer, 1982;Nishimoto et al, 1982;Madsen et al, 1983;Tartakoff, 1983a;Burditt et al, 1985). Effects on proteoglycan secretion and procollagen secretion vary a t different doses (Tajiri et al, 1980;Mitchell and Hardingham, 1982;Goldberg and Toole, 1984).…”

Section: Binding To the Rer And Nuclear Envelopementioning

confidence: 99%

Binding of lectin‐fluorescein conjugates to intracellular compartments of growth‐plate chondrocytes in situ

Farnum

1985

Am. J. Anat.

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In this study, lectin-binding techniques are applied to growth-plate cartilage to analyze the intracellular localization of lectin-binding glycoconjugates of chondrocytes in situ. The binding of ten fluorescein-conjugated lectins is analyzed on 1-micron-thick Epon-embedded, nondecalcified sections of growth plates from Yucatan swine. Comparisons are made to intracellular binding in chondrocytes of tracheal, articular, and auricular cartilage. Ear epithelium, tracheal epithelium, and kidney are used as positive control tissues for the specificity of lectin binding. Only the mannose-binding lectins had affinity for the RER and nuclear envelope. Eight lectins reacted within the Golgi complex with characteristic patterns which ranged from localized fine linear strands to extensive vesicular accumulations. When cartilage slabs were exposed before embedment to the ionophore monensin to disrupt intracellular transport through the Golgi, it was possible to define differential subcompartments of the Golgi complex, based upon sites of sugar addition. Also, it was possible to characterize the cytoplasmic deposits of reserve-zone chondrocytes which were positive with concanavalin A as glycogen, based upon their sensitivity to amylase. This method allows resolution at the light-microscopic level of lectin-binding glycoconjugates with localization to specific organelles. Patterns of intracellular binding were consistent with biochemical data relating to the subcellular localization of processing steps of complex carbohydrates prior to secretion.

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“…Thus, it was shown that heparin stimulates the synthesis and changes the sulfation pattern of the heparan sulfate from endothelial cells [Nader et al, 19891 and selenate arrests the synthesis of this compound [Dietrich et al, 19881. Monensin, a monovalent ionophore, has been shown to alter the normal structure of the Golgi complex and appears to slow or to arrest intraGolgi transport as well as to inhibit trans-Golgi functions such as terminal N-glycosylation, protein processing, and the sulfation of proteoglycans [Farquhar, 1985;Griffiths et al, 1983;Ledger and Tanzer, 1984;Tartakoff, 1982,19831. The influence of monensin on chondroitin sulfate and dermatan sulfate proteoglycan biosynthesis has been studied in chondrocytes of different origins [Burditt et al, 1985;Tanzer, 1981, 1982;Madsen et al, 1983;Nishimot0 et al, 1982a,b;Tajiri et al, 19801, rat chondrosarcoma [Mitchell and Hardingham, 19821, human skin fibroblasts [Hoppe et al, 19851, human melanoma [Bumol and Reisfel, 1982;Bumol et al, 1984;Harper et al, 19861, and rat ovarian granulosa cells wanagishita and Hascall, 19851 where it was shown a marked decrease in the incorporation of sulfate into the glycosaminoglycan chains. This undersulfation for chondroitin sulfate proteoglycan was due to a decreased 6-sulfation of the N-acetylgalactosamine moiety [Kajiwara and Tanzer, 1981;Madsen et al, 1983;Nishimoto et al, 1982a,b;Tajiri et al, 19801, whereas for dermatan sulfate proteoglycan it was related to a decrease of the 4-sulfated disaccharide containing iduronic acid residues [Hoppe et al, 1985;Yanagishita and Hascall, 19851.…”

mentioning

confidence: 99%

“…The influence of monensin on chondroitin sulfate and dermatan sulfate proteoglycan biosynthesis has been studied in chondrocytes of different origins [Burditt et al, 1985;Tanzer, 1981, 1982;Madsen et al, 1983;Nishimot0 et al, 1982a,b;Tajiri et al, 19801, rat chondrosarcoma [Mitchell and Hardingham, 19821, human skin fibroblasts [Hoppe et al, 19851, human melanoma [Bumol and Reisfel, 1982;Bumol et al, 1984;Harper et al, 19861, and rat ovarian granulosa cells wanagishita and Hascall, 19851 where it was shown a marked decrease in the incorporation of sulfate into the glycosaminoglycan chains. This undersulfation for chondroitin sulfate proteoglycan was due to a decreased 6-sulfation of the N-acetylgalactosamine moiety [Kajiwara and Tanzer, 1981;Madsen et al, 1983;Nishimoto et al, 1982a,b;Tajiri et al, 19801, whereas for dermatan sulfate proteoglycan it was related to a decrease of the 4-sulfated disaccharide containing iduronic acid residues [Hoppe et al, 1985;Yanagishita and Hascall, 19851. These studies furnished important clues as to the location of the different biosynthetic steps of chondroitin sulfate and dermatan sulfate in the Golgi apparatus.…”

mentioning

confidence: 99%

Effect of monensin on the sulfation of heparan sulfate proteoglycan from endothelial cells

Sampaio

Dietrich

Colburn

et al. 1992

J of Cellular Biochemistry

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Monensin is a monovalent metal ionophore that affects the intracellular translocation of secretory proteins at the level of trans-Golgi cisternae. Exposure of endothelial cells to monensin results in the synthesis of heparan sulfate and chondroitin sulfate with a lower degree of sulfation. The inhibition is dose dependent and affects the ratio [35S]-sulfate/[3H]-hexosamine of heparan sulfate from both cells and medium, with no changes in their molecular weight. By the use of several degradative enzymes (heparitinases, glycuronidase, and sulfatases) the fine structure of the heparan sulfate synthesized by control and monensin-treated cells was investigated. The results have shown that among the six heparan sulfate disaccharides there is a specific decrease of the ones bearing a sulfate ester at the 6-position of the glucosamine moiety. All other biosynthetic steps were not affected by monensin. The results are indicative that monensin affects the hexosamine C-6 sulfation, and that this sterification is the last step of the heparan sulfate biosynthesis and should occur at the trans-Golgi compartment.

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Synthesis and secretion of proteoglycans by cultured chondrocytes

Cited by 14 publications

References 26 publications

Monensin-induced inhibition of cell spreading in normal and dystrophic human fibroblasts.

Monensin-induced inhibition of cell spreading in normal and dystrophic human fibroblasts.

Binding of lectin‐fluorescein conjugates to intracellular compartments of growth‐plate chondrocytes in situ

Effect of monensin on the sulfation of heparan sulfate proteoglycan from endothelial cells

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