Summary
In gills of the shore crab Carcinus maenas an ATPase activity was found which was stimulated by bicarbonate and inhibited by low concentration of oligomycin and thiocyanate. This ATPase was activated by small hydrated alkali cations, i.e., activation was absent in the presence of Li+, small in the presence of Na+, and highest in the presence of K+ (K m=4 mM). Inhibitor studies using ouabain, NEM, and vanadate suggest that this ATPase is different from (Na++K+)-ATPase, the H+-ATPase of organelles, or an E 1 E 2-type ATPase represented by the H+/K+-ATPase in gastric mucosa. Results obtained by differential and density gradient centrifugation indicate that this ATPase is located in crab gill mitochondria, a location ruling out its direct participation in transepithelial ion transport. Since the ATPase lacked specific Cl--activation it is not considered to be a Cl- pump but a mitochondrial F 1 F 0-ATPase. Specific activities of mitochondrial ATPase and (Na++K+)-ATPase were of comparable magnitude. Both ATPases were greatly increased in gills of crabs acclimated to brackish water (salinity 10‰) compared to crabs maintained in sea water (30‰). These results imply that low salinity-induced modifications in branchial tissues include mechanisms for active ion uptake as well as the elements for provision of cellular energy.
Similar content being viewed by others
Abbreviations
- ATPase :
-
adenosine triphosphatase
- HEPES :
-
N-(2-hydroxyethyl)-1-piperazine-N′(2-ethanesulfonic acid)
- LDH :
-
lactate dehydrogenase
- NADH :
-
reduced nicotinamide adenine dinucleotide
- NEM :
-
Niethylmaleimide
- PEP :
-
phosphoenolpyruvate
- PK :
-
pyruvate kinase
- TRIS TRIS:
-
(hydroxymethyl)aminomethane
- S :
-
salinity
References
Allen JC, Schwartz A (1969) A possible biochemical explanation for the insensitivity of the rat to cardiac glycosides. J Pharmacol Exp Ther 168:42–46
Barra J-A, Pequeux A, Humbert W (1983) A morphological study on gills of a crab acclimated to fresh water. Tissue and Cell 15:583–596
Battram JC (1987) Chloride uptake in the gills of the fresh-water-adapted brown trout (Salmo trutta). Comp Biochem Physiol 86A:245–249
Bornancin M, DeRenzis G, Maetz J (1977) Branchial chloride transport, anion stimulated ATPase, and acid-base balance in Anguilla anguilla adapted to fresh water. Effects of hyperoxia. J Comp Physiol 117:313–322
Bornancin M, DeRenzis G, Naon R (1980) Cl-/HCO -3 -ATPase in the gills of rainbow trout: evidence for its microsomal localization. Am J Physiol 238:R251-R259
DePew EF, Towle DW (1979) Bicarbonate-stimulated ATPase in plasma membrane fractions of fiddler crab (Uca minax) gill. Mar Biol Lett 1:59–67
DeRenzis G, Bornancin M (1977) Cl-/HCO -3 -ATPase in the gills of Carassius auratus: its inhibition by thiocyanate. Biochem Biophys Acta 467:192–207
Dietz TH, Findley AM (1980) Ion-stimulated ATPase activity and NaCl uptake in the gills of freshwater mussels. Can J Zool 58:917–923
Diwan JJ (1987) Mitochondrial transport of K+ and Mg2+. Biochem Biophys Acta 895:155–165
Gereneser GA, White JF, Gradman D, Bonting SL (1988) Is there a Cl- pump? Am J Physiol 255:R677-R692
Graszynski K, Bigalke R (1987) Osmoregulation and ion transport in the extremely euryhaline fiddler crabs Uca pugilator and Uca tangeri (Ocypodidae). Zool Beitr N F 30:339–358
Henry RP (1988) Subcellular distribution of carbonic anhydrase activity in the gills of the blue crab Callinectes sapidus. J Exp Zool 245:1–8
Kerstetter TH, Kirschner LB (1974) HCO -3 -dependent ATPase activity in the gills of rainbow trout (Salmo gairdneri). Comp Biochem Physiol 48B:581–589
Kirschner LB (1979) Control mechanisms in crustaceans and fishes. In: Gilles R (ed) Mechanisms of osmoregulation in animals. John Wiley & Sons, Chichester New York Brisbane Toronto, pp 157–222
Lardy H, Reed P, Chiu Lin C-H (1975) Antibiotic inhibitors of mitochondrial ATP synthesis. Fed Proc 34:1707–1710
Lee S-H (1982) Salinity adaptation of HCO -3 -dependent ATPase activity in the gills of blue crab (Callinectes sapidus) Biochim Biophys Acta 689:143–154
Mitchell P (1961) Coupling of phosphorylation to electron and hydrogen transfer by a chemiosmotic type of mechanism. Nature 191:144–148
Mitchell P, Moyle J (1968) Proton translocation coupled to ATP hydrolysis in rat liver mitochondria. European J Biochem 4:530–539
Naon P, Bornancin M, DeRenzis G (1981) Cl-/HCO -3 -dependent ATPase in the gills of freshwater- and seawater-adapted eel (Anguilla anguilla). Biochim 63:37–43
Schuurmans Stekhoven F, Bonting SL (1981) Transport adenosine triphosphatases: properties and functions. Physiol Rev 61:1–76
Serrano R, Kanner BJ, Racker E (1976) Purification and properties of the translocating adenosine triphospatase complex of bovine heart mitochondria. J Biol Chem 251:2453–2461
Siebers D, Leweck K, Markus H, Winkler A (1982) Sodium regulation in the shore crab Carcinus maenas related to ambient salinity. 69:37–43
Siebers D, Winkler A, Lucu C, Thedens G, Weichert D (1985) Na−K-ATPase generates an active transport potential in the gills of the hyperregulating shore crab Carcinus maenas. Mar Biol 87:185–192
Siebers D, Lucu C, Winkler A (1987) Active influx of ions across the gills of osmoregulating shore crabs Carcinus maenas. Zool Beitr N F 30:315–338
Solaini G, Baracca A, Castelli GP, Rossi CA (1988) Effect of 2-hydroxy-5-nitrobenzyl bromide on proton translocation by the mitochondrial H+-ATPase. Biochem Biophys Res Comm 155:130–137
Stone DK, Xie X-S (1988) Proton translocating ATPases: Issues in structure and function. Kidney Int 33:767–774
Taylor PM, Andrews EB (1987) Tissue adenosine-triphosphatase activities in the gill and excretory system in mesogastropod molluscs in relation to osmoregulatory capacity. Comp Biochem Physiol 86A:693–696
Towle DW (1981) Role of Na+−K+-ATPase in ionic regulation by marine and estuarine animals. Mar Biol Lett 2:107–121
Towle DW (1984) Membrane-bound ATPases in arthropod ion transporting tissues. Am Zool 24:177–185
Towle DW, Hølleland T (1987) Ammonium ion substitutes for K+ in ATP-dependent Na+ transport by basolateral membrane vesicles. Am J Physiol 252:R479-R489
Van Amselvoort JMM, DePont JJHHM, Bonting SL (1977) Is there a plasma membrane-located anion-sensitive ATPase? Biochem Biophys Acta 466:283–301
Welcomme L, Devos P (1988) Cytochrome c oxidase and Na+−K+-ATPase activities in the anterior and posterior gills of the shore crab Carcinus maenas L. after adaptation to various salinities. Comp Biochem Physiol 89B:339–341
Wharton DC, Tzagoloff T (1967) Cytochrome oxidase from beef heart mitochondria. Methods Enzymology 10:245–250
Wheeler AP, Harrison EW (1982) Subcellular localisation and characterization of HCO -3 -ATPase from the mantle of the freshwater clam Anodonta cataracta. Comp Biochem Physiol 71B:629–636
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Siebers, D., Petrausch, G. & Böttcher, K. Is there a chloride ATPase in the gills of the shore crab Carcinus maenas?. J Comp Physiol B 160, 223–231 (1990). https://doi.org/10.1007/BF00300958
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00300958