Abstract
Ethane-1-hydroxy-1,1-diphosphonate (EHDP) given subcutaneously to rats in high doses (10 mg P/Kg body weight daily×10) inhibited the transport of calcium by rat duodenal gut sacs, when45Ca serosal/mucosal concentration ratios were used to measure active transport. Electron microscopic studies revealed the accumulation of electron-dense granules in the microvilli, representing a bound form of calcium and a paucity of such granules in mitochondria, changes identical to those seen in rachitic rats. These results suggest that the defect is mediated in part by interference in transport of calcium from microvilli to other intracellular sites.
Résumé
De l'éthane-1-hydroxy-1,1-diphosphonate (EHDP), administré par voie sous-cutanée à des rats à doses élevées (10 mg P/Kg de poids par jour×10), inhibe le transport du calcium au niveau du duodenum de rats, en utilisant des rapports de concentration serum/muqueuse en45Ca pour mesurer le transfert actif. Des études de microscopie électronique montrent l'accumulation de granules denses aux électrons dans les microvillosités: ces granules seraient constitués par une forme de calcium lié. On note aussi la rareté de ces granules dans les mitochondries; ces modifications sont analogues à celles observées dans les rats rachitiques. Il semble que les phénomènes pathologiques soient liées à un trouble du transport du calcium des microvillositées vers d'autres sites intracellulaires.
Zusammenfassung
Es zeigte sich, daß bei Ratten, die mit hohen Dosen (10 mg P/kg Körpergewicht täglich×10) von Aethan-1-hydroxy-1,1-diphosphonat (EHDP) subkutan behandelt wurden, der Calciumtransport im Duodenum (gut sacs) gehemmt wurde; zur Messung des aktiven Transportes wurde das45Ca-Konzentrationsverhältnis Serosa/Mucosa benützt. Elektronenmikroskopische Untersuchungen zeigten in den Microvilli eine Anhäufung von elektronendichten Granula, die eine gebundene Form von Calcium darstellen; eine kleine Anzahl solcher Granula fand sich in den Mitochondrien; diese Veränderungen können auch bei rachitischen Ratten beobachtet werden. Aus den Resultaten kann geschlossen werden, daß dieser Defekt teilweise durch eine Interferenz des Calciumtransportes von den Microvilli zu anderen intrazellulären Stellen bedingt ist.
Similar content being viewed by others
References
Bonjour, J. P., Herrath, D., Schaefer, K., Fleisch, H.: Calcium absorption and vitamin D metabolism in diphosphonate treated rats. Experientia (Basel)28, 726 (1972)
Bonjour, J., Russell, R. G. G., Morgan, D. B., Fleisch, H. A.: Intestinal calcium absorption, Ca-binding protein, and Ca-ATPase in diphosphonate-treated rats. Amer. J. Physiol.224, 1011–1017 (1973)
Elder, J. A., Schraer, H.: Calcium uptake by mitochondria from the avian shell gland, Fed. Proc.28, 284 (1969)
Fleisch, H., Russell, R. G. G., Bisaz, S., Casey, P. A., Müehlbauer, R. C.: The inhibitory effect of phosphonates on the formation of calcium phosphate crystals in vitro and on aortic and kidney calcification in vivo. Europ. J. clin. Invest.1, 12–18 (1970)
Fleisch, H., Russell, R. G. G., Francis, M. D.: Diphosphonates inhibit hydroxyapatite dissolution in vitro and bone resorption in tissue culture and in vivo. Science165, 1262–1264 (1969)
Francis, M. D.: The inhibition of calcium hydroxyapatite crystal growth. Calcif. Tiss. Res.3, 151–162 (1969)
Gasser, A. B., Morgan, D. B., Fleisch, H. A.: The influence of two diphosphonates on calcium metabolism in the rat. Clin. Sci.43, 31–45 (1972)
Gomori, G. A.: Modification of the colorimetric phosphorus determination for use with the photoelectric colorimeter. Lab. Clin. Med.27, 955–960 (1942)
Gustafson, R. A., Thurston, E. L.: Calcium deposition in the myxomycete didymium squamalosum. Tex. Rep. Biol. Med. (in press)
Hamilton, J. W., Holdsworth, E. S.: The release of45Ca from mitochondria of chicken intestinal mucosa by calcium binding protein. Biochem. biophys. Res. Commun.40, 1315–1330 (1970)
Krawitt, E. L.: The role of intestinal transport proteins in cortisone mediated suppression of calcium absorption. Biochim. biophys. Acta (Amst.)274, 179–188 (1972)
Krawitt, E. L., Schedl, H. P.: In vivo calcium transport by rat small intestine. Amer. J. Physiol.214, 232–236 (1968)
Martin, D. L., DeLuca, H. F.: Calcium transport and the role of vitamin D. Arch. Biochem. Biophys.134, 139–148 (1969)
Martin, J. H.: Personal communication
Martin, J. H., Matthews, J. L.: Mitochondrial granules in chondrocytes. Calcif. Tiss. Res.3, 184–193 (1969)
Matthews, J. L., Martin, J. H., Arsenis, C., Eisenstein, R., Kuettner, K.: The role of mitochondria in intracellular calcium regulation. Cellular mechanisms for calcium transfer and homeostasis (G. Nichols, Jr. and R. H. Wasserman. eds.) p. 239–255. New York: Academic Press 1971
Matthews, J. L., Martin, J. H., Sampson, H. W., Kunin, A. S., Roan, J. H.: Mitochondrial granules in the normal and rachitic rat epiphysis. Calcif. Tiss. Res.5, 91–99 (1970)
Meyer, W. L., Kunin, A. S.: The inductive effect of rickets on glycolytic enzymes of rat epiphysial cartilage and its reversal by vitamin D and phosphate. Arch. Biochem. Biophys.129, 438–446 (1969)
Michael, W. R., King, W. R., Francis, M. D.: Effectiveness of diphosphonates in preventing “osteoporosis” of disuse in the rat. Clin. Orthop.78, 271–276 (1971)
Miller, R. G., Jr.: Simultaneous statistical inference p. 272. New York: McGraw Hill 1966
Morgan, D. B., Bonjour, J., Gasser, A. B., O'Brien, K., Fleisch, H. A.: The influence of a diphosphonate on the intestinal absorption of calcium. Israel J. med. Sci.7, 384–386 (1971)
Mühlbauer, R. C., Russell, R. G. G., Williams D. A., Fleisch, H.: The effects of diphosphonates, polyphosphates, and calcitonin on “immobilisation osteoparosis” in rats. Europ. J. clin. Invest.1, 336–344 (1971)
Norman, A. W., Mircheff, A. K., Adams, T. H., Spielvogei, A.: Studies on the mechanism of the action of calciferol. III. Vitamin D-mediated increase of intestinal brush border alkaline phosphatase activity. Biochim. biophys. Acta (Amst.)215, 348–359 (1970)
Papworth, D. G., Patrick, G.: The kinetics of influx of calcium and strontium into rat intestine in vitro. J. Physiol. (Lond.)210, 999–1020 (1970)
Peachy, L. D.: Electron microscopic observation on the accumulation of divalent cations in intramitochondrial granules. J. Cell Biol.20, 95–109 (1964)
Reynolds, E. S.: Liver parenchymal cell injury. III. The nature of calcium associated electron opaque masses in rat liver mitochondria following poisoning with carbon tetrachloride. J. Cell Biol.25, 53–57 (1965)
Roth, S. L., Wyw, A. U., Kunin, A. S.: Effect of dietary deficiency on vitamin D, Calcium and phosphorus on the ultrastructure of the rat parathyroid gland. Amer. J. Path.53, 631–650 (1968)
Russell, R. G., Monda, A., Bonjour, J. P., Fleisch, H.: Relation between alkaline phosphatase and Ca-ATPase in calcium transport. Nature (Lond.) New Biol.240, 126–127 (1972)
Russell, R. G. G., Mühlbauer, R. C., Bisaz, S., Williams, D. A., Fleisch, H.: The influence of pyrophosphate, condensed phosphates, phosphonates and other phosphate compounds on the dissolution of hydroxyapatite in vitro and on bone resorption induced by parathyroid hormone in tissue culture and in thyroparathyroidectomized rats. Calcif. Tiss. Res.6, 183–196 (1970)
Sampson, H. W., Matthews, J. L., Martin, J. H., Kunin, A. S.: An electron microscopic localization of calcium in the small intestine of normal, rachitic and vitamin-D-treated rats. Calcif. Tiss. Res.5, 305–316 (1970)
Schachter, D., Dowdle, E. B., Schenker, H.: Active transport of calcium by the small intestine of the rat. Amer. J. Physiol.198, 263–268 (1960)
Schachter, D., Kowarski, S., Finkelstein, J. D., Wang Ma, R.: Tissue concentration differences during active transport of calcium by intestine. Amer. J. Physiol.211, 1131–1136 (1966)
Schenk, R., Merz, W. A., Fleisch, H.: Effect of ethane-1-hydroxy-1, 1-diphosphonate (EHDP) and dichloromethylene diphosphonate (Cl2MDP) on the calcification and resorption of cartilage and bone in the tibial epiphysis and metaphysis of rats. Calcif. Tiss. Res. (in press)
Simmons, D. J., Kunin, A. S.: Development and healing of rickets in rats. Clin. Orthop.68, 251–260 (1970)
Sokal, R. P., Rohls, F. T.: Biometry, p. 212. San Francisco: W. H. Freeman 1969
Taylor, C. M., Mauer, E. B., Reeve, A.: The metabolism of cholecalciferol (vitamin D3) in 1-hydroxyethydiphosphonate-treated chicks. Biochem. Soc. Trans.1, 44–47 (1973)
Vasington, F. D., Murphy, J. V.: Calcium uptake by rat kidney mitochondria and its dependence on respiration and phosphorylation. J. biol. Chem.237, 2670–2681 (1962)
Wasserman, R. H., Taylor, A. N.: Some aspects of the intestinal absorption of calcium with special reference to vitamin D. Mineral metabolism—an advanced treatise (C. L. Comar and F. Bronner, eds.), vol. III., p. 321. New York: Academic Press 1969
Wasserman, R. H., Taylor, A. N.: Vitamin D3-induced calcium binding protein in chick intestinal mucosa. Science152, 791 (1966)
Wilson, T. H., Wiseman, G.: The use of sacs of everted small intestine for the study of transference of substances from the mucosal to the serosal surface. J. Physiol. (Lond.)213, 116–125 (1954)
Author information
Authors and Affiliations
Additional information
Baylor College of Dentistry
Rights and permissions
About this article
Cite this article
Krawitt, E.L., Sampson, H.W., Kunin, A.S. et al. Effect of ethane-1-hydroxy-1,1-diphosphonate (EHDP) administration on duodenal calcium transport. Calc. Tis Res. 15, 21–29 (1974). https://doi.org/10.1007/BF02059040
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02059040