ON THE MECHANISMS OF BONE RESORPTION

Bone resorption, characterized by the solubilization of both the mineral and the organic components of the osseous matrix, was obtained in tissue culture under the action of parathyroid hormone (PTH). It was accompanied by the excretion of six lysosomal acid hydrolases, which was in good correlation with the progress of the resorption evaluated by the release of phosphate, calcium 45 or hydroxyproline from the explants; there was no increased excretion of two nonlysosomal enzymes, alkaline phosphatase, and catalase. Balance studies and experiments with inhibitors of protein synthesis indicated that the intracellular stores of the acid hydrolases excreted were maintained by new synthesis. The release was not due to a direct disruption of the lysosomal membrane by PTH; it is presumed to result from an exocytosis of the whole lysosomal content and to involve mechanisms similar to those controlling the secretion of this content into digestive vacuoles. The resorbing explants acidified their culture fluids at a faster rate and released more lactate and citrate than the controls; this release was in good correlation, in the PTH-treated cultures, with the resorption of the bone mineral, but the amount of citrate released was considerably smaller than that of lactate. The acid released could account for the resorption of the mineral. It is proposed, as a working hypothesis, that the acid hydrolases of the lysosomes are active in the resorption of the organic matrix of bone and that acid, originating possibly from the stimulation of glycolysis, cares for the concomitant solubilization of bone mineral while also favoring the hydrolytic action of the lysosomal enzymes.

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H+ stimulation of cell-mediated bone resorption in tissue culture

AJP Endocrinology and Metabolism ◽

10.1152/ajpendo.1987.253.1.e90 ◽

1987 ◽

Vol 253 (1) ◽

pp. E90-E98 ◽

Cited By ~ 9

Author(s):

P. Goldhaber ◽

L. Rabadjija

Keyword(s):

Tissue Culture ◽

Calcium Release ◽

Bone Destruction ◽

Calcium Content ◽

Organic Matrix ◽

Defined Medium ◽

Chemically Defined Medium ◽

The Media ◽

Neonatal Mouse Calvaria ◽

Stimulation Of

The addition of protons in the form of hydrochloric acid (10.5, 17.2, or 26.6 meq/l resulting in an initial media pH of 7.28, 7.15, and 6.94, respectively) to neonatal mouse calvaria maintained in a chemically defined medium in tissue culture for 1 wk increased calcium release in a dose-response fashion. The same amounts of protons added to the media of devitalized calvaria caused no increase in calcium release into the medium. The net cell-mediated calcium release resulting from the addition of 26.6 meq/l of protons amounted to approximately 50% of the initial calvarial calcium content. Hydroxyproline determinations revealed that active resorption was taking place, wherein both mineral and organic matrix are removed simultaneously. Histological examination of the extensively resorbed calvaria demonstrated the presence of numerous osteoclasts in different stages of bone destruction. The addition of indomethacin (100 ng/ml) strongly inhibited the increase in calcium release by added protons, suggesting that prostaglandin synthesis is involved in the phenomenon. The addition of thyrocalcitonin also inhibited proton-induced calcium release, providing additional evidence that the calcium release from cultures exposed to added protons involved osteoclastic activity.

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Fibroblast receptor for lysosomal enzymes mediates pinocytosis of multivalent phosphomannan fragment

The Journal of Cell Biology ◽

10.1083/jcb.84.1.77 ◽

1980 ◽

Vol 84 (1) ◽

pp. 77-86 ◽

Cited By ~ 45

Author(s):

HD Fischer ◽

M Natowicz ◽

WS Sly ◽

RK Bretthauer

Keyword(s):

Molecular Weight ◽

Alkaline Phosphatase ◽

Lysosomal Enzymes ◽

Human Fibroblasts ◽

Recognition System ◽

High Uptake ◽

Acid Hydrolases ◽

Phosphatase Treatment ◽

Hydrolysis Of ◽

Endoglycosidase H

Mild acid hydrolysis of phosphomannan secreted by the yeast hansenula holstii (NRRL Y- 2448) produces two phosphomannyl fragments which differ strikingly in their potency as inhibitors of pinocytosis of human β-glucuronidase by human fibroblasts. The larger molecular weight polyphosphomonoester fragment is 100,000-fold more potent an inhibitor of enzyme uptake than the smaller penta-mannosyl-monophosphate fragment. Binding to attached fibroblasts at 3 degrees C was much greater with the polyphosphomonoester fragment than with the pentamannosyl-monophosphate. The larger molecular weight fragment was also subject to adsorptive pinocytosis and was taken up by fibroblasts at a rate 30- fold greater than the rate of uptake of pentamannosyl-monophosphate. Evidence that the polyphosphomonoester fragment is taken up by the phosphomannosyl-recognition system that mediates uptake of lysosomal enzymes includes: (a) its pinocytosis is inhibited by the same compounds that competitively inhibit enzyme pinocytosis (mannose-6-phosphate and phosphomannan from saccharomyces cerevisiae mutant mnn-1); (b) alkaline phosphatase treatment greatly reduces its susceptibility to pinocytosis; (c) its pinocytosis is competitively inhibited by high-uptake human β-glucuronidase; and (d) this inhibition by high-uptake enzyme is dramatically reduced by prior treatment of the enzyme with alkaline phosphatase or endoglycosidase-H. Endoglycosidase-H treatment human β-glucuronidase dramatically reduced its susceptibility to pinocytosis by fibroblasts. The phosphomannosyl components of high- uptake enzyme released by endoglycosidase-H treatment were much less effective inhibitors of polyphosphomonoester pinocytosis than when present on the phosphomannyl-enzyme. These results suggest that high-uptake acid hydrolases may be polyvalent ligands analogous to the polyphosphomonoester mannan fragment whose pinocytosis depends on interaction of more than one phospho-mannosyl recognition marker with pinocytosis receptors on fibroblasts.

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