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Mitochondrial ATPase in the gills of the shore crabCarcinus maenas
Helgoländer Meeresuntersuchungen volume 46, pages 435–445 (1992)
Abstract
Posterior gills (No. 7 and 8) of shore crabsCarcinus maenas were homogenized and fractionated by means of differential and density gradient centrifugation. Employment of marker enzymes Na-K-ATPase and carbonic anhydrase for plasma membranes and cytochrome oxidase for mitochondria showed that these structural elements were separated. Ultramicroscopic investigations of combined fractions confirmed the presence of the respective mitochondrial and vesicular plasma membrane structures. An ATPase which did not depend on the presence of sodium (20 mM) ions in the incubation medium but on the presence of potassium (20 mM) ions only was found in the mitochondrial fractions. The mitochondrial ATPase was tightly bound to cellular particulates and activated approximately threefold by bicarbonate (20 mM) ions. The activity of this ATPase was nearly completely inhibited by oligomycin (1 μg ml−1) and greatly inhibited by low levels (5 mM) of thiocyanate and calcium ions, the Ki for Ca2+ being ca 4 mM. The results obtained confirm literature data on high mitochondrial densities in crab gills and allow the assumption of significant rates of energy metabolism in these organs. Considering its properties the mitochondrial ATPase is clearly distinct from crab gill Na-K-ATPase and can be measured specifically in samples containing Na-K-ATPase. Mitochondrial ATPase is therefore considered a suitable and reliable marker enzyme for mitochondria.
Literature Cited
Burnett, L. E. & McMahon, B. R., 1985. Facilitation of CO2 excretion by carbonic anhydrase located in the surface of the basal membrane of crab gill epithelium. — Resp. Physiol.62, 341–348.
Böttcher, K., Siebers, D., Becker, W. & Petrausch, G., 1991. Physiological role of branchial carbonic anhydrase in the shore crabCarcinus maenas. — Mar. Biol.,110, 337–342.
Bruns, W., Dermietzel, R. & Gros, G., 1986. Carbonic anhydrase in the sarcoplasmic reticulum of rabbit skeletal muscle. — J. Physiol.371, 351–364.
Copeland, D. E. & Fitzjarrel, A. T., 1968. The salt-absorbing cells in the gills of the blue crab (Callinectes sapidus Rathbun) with notes on modified mitochondria. — Z. Zellforsch. mikrosk. Anat.92, 1–22.
Ebel, R. E. & Lardy, H. A., 1975. Stimulation of rat liver mitochondrial adenosine triphosphatase by anions. — J. biol. Chem.250, 191–196.
Goodman, S. H. & Cavey, M. J., 1990. Organization of a phyllobranchiate gill from the green shore crabCarcinus maenas (Crustacea, Decapoda). — Cell Tissue Res.,260, 495–505.
Graszynski, K., 1970. Intrazelluläre Lokalisation von Enzymen des Flußkrebses. — Z. vergl. Physiol.66, 107–122.
Kirschner, L. B., 1979. Control mechanisms of crustaceans and fishes. In: Mechanisms of osmoregulation in animals. Ed. by R. Gilles, Wiley, Chichester, 157–222.
Kormanik, G. A. & Cameron, J. N., 1981. Ammonia excretion in animals that breath water: A review. — Mar. Biol. Lett.2, 11–23.
Lardy, H., Reed, P. & Chiu Lin, C.-H., 1975. Antibiotic inhibitors of mitochondrial ATP synthesis. —Fedn Proc. Am. Socs exp. Biol.34, 1707–1710.
Lucu, C., 1990. Ionic regulatory mechanisms in crustacean gill epithelia. — Comp. Biochem. Physiol.97A, 297–306.
Maren, T. H., Ash, V. I. & Bailey, Jr., 1954. Carbonic anhydrase inhibition. — Bull. Johns Hopkins Hosp.95, 244–255.
Maren, T. H., 1960. A simplified micro method for the determination of carbonic anhydrase and its inhibitors. — J. Pharmac. exp. Ther.130, 26–29.
Oglesby, L. C., 1981. Volume regulation in aquatic invertebrates. — J. exp. Zool.215, 289–301.
Péqueux, A., Chapelle, S., Wanson, S., Goffinet, G. & Francois, C., 1983. (Na++K+) ATPase activity and phospholipid content of various fractions of the posterior gills ofCarcinus maenas andEriocheir sinensis. — Mar. Biol. Lett.4, 267–279.
Perry, S. F. & Laurent, P., 1990. The role of carbonic anhydrase in carbon dioxide excretion, acidbase-balance and ionic regulation in aquatic gill breathers. In: Comparative Physiology. Ed. by R. K. H. Kinne, E. Kinne-Saffran & K. W. Beyenbach. Karger, Basel,2, 39–57.
Siebers, D., Leweck, K., Markus, H. & Winkler, A., 1982. Sodium regulation in the shore crabCarcinus maenas as related to ambient salinity. — Mar. Biol.69, 37–43.
Siebers, D., Winkler, A., Leweck, K. & Madian, A., 1983. Regulation of sodium in the shore crabCarcinus maenas adapted to environments of constant and changing salinities. — Helgoländer Meeresunters.36, 303–312.
Siebers, D., Petrausch, G. & Böttcher, K., 1990. Is there a chloride ATPase in the gills of the shore crabCarcinus maenas?. J. comp. Physiol. (B)160, 223–231.
Towle, D. W., 1981. Na+-K+-ATPase in ionic regulation by marine and estuarine animals. — Mar. Biol. Lett.2, 107–121.
Truchot, J. P., 1979. Mechanisms of the compensation of blood respiratory acid-base disturbances in the shore crab,Carcinus maenas (L.). — J. exp. Zool.210, 407–416.
Wharton, D. C. & Tzagoloff, A., 1967. Cytochrome oxidase from beef heart mitochondria. — Meth. Enzym.10, 245–250.
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Siebers, D., Hentschel, J., Böttcher, K. et al. Mitochondrial ATPase in the gills of the shore crabCarcinus maenas . Helgolander Meeresunters 46, 435–445 (1992). https://doi.org/10.1007/BF02367209
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DOI: https://doi.org/10.1007/BF02367209