Summary
Liver slices incubated at 1 °C underwent swelling of both cellular and intercellular compartments, as judged by electronmicroscopy. The ultrastructure showed marked changes, including disorganization of the cytocavitary network and plasma membrane and alterations of mitochondria. Restoration of metabolically favorable conditions (oxygenated medium at 38 °C) caused a nearly complete recovery of ultrastructure closely associated with extrusion of water; measurements of inulin space and electronmicroscopy both indicate a recovery of cell volume, with intercellular spaces remaining somewhat expended. The fluid lost was a roughly isotonic solution of Na+ and Cl−, while K+ was reaccumulated in exchange for Na+. Cyanide prevented recovery. Ouabain and oligomycin each partially prevented fluid extrusion, but had little effect on ultrastructural recovery except to induce intracellular vesicles containing particles of thorium dioxide derived from sinusoidal spaces. The vesicles were, however, markedly different in form with each inhibitor. There are, thus ouabain-sensitive and-insensitive components of volume regulation; the former appears to depend on the coupled transport of Na+ and K+ and the latter, we suggest, on a secretion of Na+ and Cl− into vesicles which release their contents into the bile canaliculi by an oligomycin-sensitive mechanism. Mitochondria showed conformational changes between orthodox and condensed forms, but these could not be directly related to tissue energy states; the numbers of mitochondrial dense granules bore a closer relation to tissue ATP.
Similar content being viewed by others
References
Bakkeren, J.A.J.M., Bonting, S.L. 1968. Studies on (Na+−K+)-activated ATPase. XX. Properties of (Na+−K+)-activated ATPase in rat liver.Biochim. Biophys. Acta 150:460
Bakkeren, J.A.J.M., Bonting, S.L. 1968. Studies on (Na+−K+)-activated ATPase. XXI. Changes in (Na+−K+)-activated ATPase activity and ouabain-sensitive86Rb+ uptake rate in regenerating rat liver.Biochim. Biophys. Acta 150:467
Chance, B., Williams, G.R. 1955. Respiratory enzymes in oxidative phosphorylation. III. The steady state.J. Biol. Chem. 217:409
Deyrup, I. 1953. Reversal of fluid uptake by rat kidney slices immersed in isosmotic solutionsin vitro.Am. J. Physiol. 175:349
Elshove, A., Van Rossum, G.D.V. 1963. Net movements of sodium and potassium, and their relation to respiration, in slices of rat liver incubatedin vitro.J. Physiol. (London) 168:531
Emmelot, P., Bos, C.J. 1966. Studies on plasma membranes. III. Mg2+-ATPase, (Na2+−K+−Mg2+)-ATPase and 5′-nucleotidase activity of plasma membranes isolated fromrat liver.Biochim. Biophys. Acta 120:369
Emmelot, P., Bos, C.J., Benedetti, E.L., Rümke, P. 1964. Studies on plasma membranes. I. Chemical composition and enzyme content of plasma membranes isolated from rat liver.Biochim. Biophys. Acta 90:126
Ginn, F.L., Shelburne, J.D., Trump, B.F. 1968. Disorders of cell volume regulation. I. Effects of inhibition of plasma membrane adenosine triphosphatase with ouabain.Am. J. Pathol. 53:1041
Hackenbrock, C.R. 1966. Ultrastructural bases for metabolically linked mechanical activity in mitochondria. I. Reversible ultrastructural changes with change in metabolic steady state in isolated liver mitochondria.J. Cell. Biol. 30:269
Hackenbrock, C.R. 1968. Ultrastructural bases for metabolically linked mechanical activity in mitochondria. II. Electron transport-linked ultrastructural transformations in mitochondria.J. Cell. Biol. 37:345
Heckmann, K.D., Parsons, D.S. 1959. Changes in the water and electrolyte content of rat-liver slicesin vitro.Biochim. Biophys. Acta 36:203
Heckmann, K.D., Parsons, D.S. 1959. Electrolyte distribution between rat-liver slices and an artificial saline medium.Biochim. Biophys. Acta 36:213
Judah, J.D., Ahmed, K. 1964. Inhibitions of transport and cation activated ATPases.J. Cell. Comp. Physiol. 64:355
Judah, J.D., McLean, A.E.M. 1962. Action of antihistamine drugsin vitro. — II. Ion movements and phosphoproteins in whole cells.Biochem. Pharmacol. 11:593
Kleinzeller, A. 1972. Cellular transport of water. L.E. Hokin, editor.In: “Metabolic Pathways”. 3rd Ed., Vol. VI, pp. 91–131. Academic Press, New York
Leaf, A. 1956. On the mechanism of fluid exchange of tissuesin vitro.Biochem. J. 62:241
MacKnight, A.D.C., Pilgrim, J.P., Robinson, B.A. 1974. The regulation of cellular volume in liver slices.J. Physiol. (London) 238:279
Millonig, G. 1961. A modified procedure for lead staining of thin sections.J. Biophys. Biochem. Cytol. 11:736
Parsons, D.S., Van Rossum, G.D.V. 1962. The effects of animal age on the swelling of rat liver slicesin vitro.Q. J. Exp. Physiol. 47:39
Parsons, D.S., Van Rossum, G.D.V. 1962. Observations on the size of the fluid compartments of rat liver slices in vitro. J. Physiol. (London)164:116
Robinson, J.R. 1950. Osmoregulation in surviving slices from the kidneys of adult rats.Proc. R. Soc. London B. 137:378
Rouiller, C., Matter, A., Orci, L. 1969. A study on the permeability barriers between Disse's space and the bile canaliculus.J. Ultrastruct. Res. Suppl.11
Schwartz, A. 1964. A Na++K+-stimulated adenosine triphosphatase in “microsomal” fractions from rat liver.Biochim. Biophys. Acta 67:329
Thurman, R.G., Scholz, R. 1969. Mixed function oxidation in perfused rat liver. The effect of aminopyrine on oxygen uptake.Eur. J. Biochem. 10:459
Trump, B.F., Bulger, R.E. 1971. Experimental modification of lateral and basilar plasma membranes and extracellular compartments in the flounder nephron.Fed. Proc. 30:22
Van Rossum, G.D.V. 1964. The effect of oligomycin on net movements of sodium and potassium in mammalian cellsin vitro.Biochim. Biophys. Acta 82:556
Van Rossum, G.D.V. 1966. Effects of potassium, ouabain and valinomycin on the efflux of24Na+ and pyridine nucleotides of rat-liver slices.Biochim. Biophys. Acta 122:323
Van Rossum, G.D.V. 1970. On the coupling of respiration to cation transport in slices of rat liver.Biochim. Biophys. Acta 205:7
Van Rossum, G.D.V. 1970. Relation of intracellular Ca2+ to retention of K+ by liver slices.Nature (London) 225:638
Van Rossum, G.D.V. 1970. Net movements of calcium and magnesium in slices of rat liver.J. Gen. Physiol. 55:18
Van Rossum, G.D.V. 1976. The effects of oligomycin on energy metabolism and cation transport in slices of rat liver. Inhibition of oxidative phosphorylation as the primary action.Biochim. Biophys. Acta 423:111
Van Rossum, G.D.V. 1972. The relation of sodium and potassium ion transport to the respiration and adenine nucleotide content of liver slices treated with inhibitors of respiration.Biochem. J. 129:427
Whittam, R. 1971. Active cation transport as a pacemaker of respiration.Nature (London) 191:603
Whittembury, G., Proverbio, F. 1970. Two modes of Na extrusion in cells from Guinea pig kidney cortex slices.Pfluegers Arch. 316:1
Wilson, T.H. 1954. Ionic permeability and osmotic swelling of cells.Science 120:104
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Russo, M.A., van Rossum, G.D.V. & Galeotti, T. Observations on the regulation of cell volume and metabolic controlin vitro; changes in the composition and ultrastructure of liver slices under conditions of varying metabolic and transporting activity. J. Membrain Biol. 31, 267–299 (1977). https://doi.org/10.1007/BF01869409
Received:
Revised:
Published:
Issue Date:
DOI: https://doi.org/10.1007/BF01869409