Ultrastructural cytochemistry of complex carbohydrates in osteoblasts, osteoid, and bone matrix

Takagi, Minoru; Parmley, Richard T.; Toda, Yoshihisa; Denys, Francis R.

doi:10.1007/bf02405052

Cited by 27 publications

(15 citation statements)

References 39 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…(a) xi5 000, (b) x37 500. electron-microscopical level or have used mAbs immunohistochemically specific for PGs (Poole et al, 1986;Bianco et al, 1990;Takagi et al, 1991). Some of these studies suggest either a loss of, or a sharp decrease in, PG staining'at the mineralization front (Baylink et al, 1972;Wergedal & Baylink, 1974;Takagi et al, 1983Takagi et al, , 1991. However, the latter immunohistochemical studies have not been extended to the ultrastructural level as was done in the present study.…”

Section: Resultsmentioning

confidence: 57%

“…(1982), using mAbs specific for cartilage PG monomer core protein and link protein, demonstrating heavy immunostaining in mineralized cartilage of bovine growth plate. Similar ultrastructural studies using cationic dye-binding methods have localized strong sulphate staining in mineralized cartilage matrix (Takagi et al, 1983;Scherft & Moskalewski, 1984;., 1991). The lack of 5-D-4 (KS) staining in mineralized cartilage matrix is in agreement with previous biochemical studies demonstrating that large aggregating PGs from rat cartilage contain little or no KS, since the protein region rich in KS is not present in this specie), (Antonsson et al, 1989;Heineg~rd et aI., 1990).…”

Section: Resultsmentioning

confidence: 93%

“…In prebone, there was fine The present study has utilized ultrastructural immunocytochemical methods specific for C0-S, C4-S, DS, C6-S and KS to demonstrate the type and localization of GAGs associated with mineralization of bone and cartilage matrix. Previous studies examining PGs in prebone and/or bone matrix have used either cationic dye-binding methods (Schefft, 1968;Bernald & Pease, 1969;Bonucci, 1971;Baylink et al, 1972;Groot, 1982a;Takagi et al, 1983;Sauren et al, 1989Sauren et al, , 1991, or electron probe analytical methods (Wergedal & Baylink, 1974;Groot, 1982b;Arsenault & Ottensmeyer, 1984), at the light-and Fig. 1.…”

Section: Resultsmentioning

confidence: 99%

“…Light and electron microscopical studies using a cationic dye-binding technique and electron probe analysis have demonstrated that the content of sulphate and/or sulphur is high 'in prebone (Baylink et al, 1972;Wergedal & Baylink, 1974;Groot, 1982a, b;Takagi et aI., 1983;Sauren et al, 1989Sauren et al, , 1991, but drops abruptly at the mineralization front (Baylink et at., 1972;Wergedal & Baylink, 1974;Takagi et al, 1983). Light microscopical studies of metaphyseal bone trabeculae of fetal bovine fetus, using an antibody specific for small dermatan sulphate (DS) PGs of cartilage, have shown immunohistochemically that the antibody stains only prebone---and not the mineralized bone matrix (Poole et al, 1986).…”

Section: Introductionmentioning

confidence: 98%

See 3 more Smart Citations

Immunoelectron microscopic studies of glycosaminoglycans in the metaphyseal bone trabeculae of growing rats

et al. 1992

Self Cite

View full text Add to dashboard Cite

The types and distribution of glycosaminoglycans (GAGs) were studied immunocytochemically in osteoid, mineralized bone matrix, and cartilage matrix of growing rat metaphyseal bone after aldehyde fixation and EDTA demineralization, using four monoclonal antibodies (mAbs 1-B-5, 2-B-6, 3-B-3 and 5-D-4). These mAbs specifically recognize epitopes in non-sulphated chondroitin (C0-S); chondroitin 4-sulphate (C4-S) and dermatan sulphate (DS); chondroitin 6-sulphate (C6-S) and C0-S; and keratan sulphate (KS) respectively. In osteoid, all mAbs except 1-B-5 weakly stained matrix material on and between collagen fibrils, and moderately stained organic material corresponding to bone nodules, which are known sites of mineralization. However, the staining of osteoid abruptly decreased at the mineralization front; weak staining was confined mostly to the organic material of bone nodules in mineralized bone matrix, with very weak or no staining of the rest of the bone matrix. This staining progressively decreased toward the mineralized cartilage matrix and became negative. The mineralized cartilage matrix and lamina limitans reacted strongly with all mAbs except 5-D-4. These results indicate that osteoid contains sulphated proteoglycans containing C4-S and/or DS, C6-S and KS, and subsequent bone matrix mineralization appears to require accumulation of these macromolecules within bone nodules and eventual loss of these substances for complete mineralization, whereas proteoglycans containing C0-S, C4-S and/or DS, and C6-S still exist in mineralized cartilage matrix and lamina limitans.

show abstract

Section: Resultsmentioning

confidence: 57%

Section: Resultsmentioning

confidence: 93%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 98%

See 2 more Smart Citations

Immunoelectron microscopic studies of glycosaminoglycans in the metaphyseal bone trabeculae of growing rats

et al. 1992

Self Cite

View full text Add to dashboard Cite

show abstract

“…A requisite corollary for such a role is presumed to be a close physical association with the earliest sites of mineralization. Using histochemical methods, Bonucci and others [Bonucci, 1967;Groot, 1982;Takagi, et al, 1983;Bonucci et al, 1988] demonstrated previously the existence of an acidic, filamentous, glycoproteinaceous phase (e.g., crystal ghost) in early ''mineralization nodules'' which shares the same shape, arrangement, and orientation as the inorganic crystallites. Recent immunocytochemical studies have documented in detail the precise distribution of a number of noncollagenous extracellular matrix and plasma proteins in relation to the mineral phase and collagen of bone [Bianco et al, 1993;Ingram et al, 1993;McKee and Nanci, 1995].…”

Section: Discussionmentioning

confidence: 98%

Bone acidic glycoprotein-75 self-associates to form macromolecular complexes in vitro and in vivo with the potential to sequester phosphate ions

et al. 1997

View full text Add to dashboard Cite

Monoclonal antibody HTP IV-#1 specifically recognizes a complexation-dependent neoepitope on bone acidic glycoprotein-75 (BAG-75) and a Mr = 50 kDa fragment. Complexes of BAG-75 exist in situ, as shown by immunofluorescent staining of the primary spongiosa of rat tibial metaphysis and osteosarcoma cell micromass cultures with monoclonal antibody HTP IV-#1. Incorporation of BAG-75 into complexes by newborn growth plate and calvarial tissues was confirmed with a second, anti-BAG-75 peptide antibody (#503). Newly synthesized BAG-75 immunoprecipitated from mineralizing explant cultures of bone was present entirely in large macromolecular complexes, while immunoprecipitates from monolayer cultures of osteoblastic cells were previously shown to contain only monomeric Mr = 75 kDa BAG-75 and a 50 kDa fragment. Purified BAG-75 self-associated in vitro to form large spherical aggregate structures composed of a meshwork of 10 nm diameter fibrils. These structures have the capacity to sequester large amounts of phosphate ions as evidenced by X-ray microanalysis and by the fact that purified BAG-75 preparations, even after extensive dialysis against water, retained phosphate ions in concentrations more than 1,000-fold higher than can be accounted for by exchange calculations or by electrostatic binding. The ultrastructural distribution of immunogold-labeled BAG-75 in the primary spongiosa underlying the rat growth plate is distinct from that for other acidic phosphoproteins, osteopontin and bone sialoprotein. We conclude that BAG-75 self-associates in vitro and in vivo into microfibrillar complexes which are specifically recognized by monoclonal antibody HTP IV-#1. This propensity to self-associate into macromolecular complexes is not shared with acidic phosphoproteins osteopontin and bone sialoprotein. We hypothesize that an extracellular electronegative network of macromolecular BAG-75 complexes could serve an organizational role in forming bone or as a barrier restricting local diffusion of phosphate ions.

show abstract

Bone cell biology: The regulation of development, structure, and function in the skeleton

1988

View full text Add to dashboard Cite

Bone cells compose a population of cells of heterogeneous origin but restricted function with respect to matrix formation, mineralization, and resorption. The local, mesenchymal origin of the cells which form the skeleton contrasts with their extraskeletal, hemopoietic relatives under which bone resorption takes place. However, the functions of these two diverse populations are remarkably related and interdependent. Bone cell regulation, presently in its infancy, is a complicated cascade involving a plethora of local and systemic factors, including some components of the skeletal matrices and other organ systems. Thus, any understanding of bone cell regulation is a key ingredient in understanding not only the development, maintenance, and repair of the skeleton but also the prevention and treatment of skeletal disorders.

show abstract

Ultrastructural cytochemistry of complex carbohydrates in osteoblasts, osteoid, and bone matrix

Cited by 27 publications

References 39 publications

Immunoelectron microscopic studies of glycosaminoglycans in the metaphyseal bone trabeculae of growing rats

Immunoelectron microscopic studies of glycosaminoglycans in the metaphyseal bone trabeculae of growing rats

Bone acidic glycoprotein-75 self-associates to form macromolecular complexes in vitro and in vivo with the potential to sequester phosphate ions

Bone cell biology: The regulation of development, structure, and function in the skeleton

Contact Info

Product

Resources

About