Abstract
Cell swelling is now admitted as being a new principle of metabolic control but little is known about the energetics of cell swelling. We have studied the influence of hypo- or hyperosmolarity on both isolated hepatocytes and isolated rat liver mitochondria. Cytosolic hypoosmolarity on isolated hepatocytes induces an increase in matricial volume and does not affect the myxothiazol sensitive respiratory rate while the absolute value of the overall thermodynamic driving force over the electron transport chain increases. This points to an increase in kinetic control upstream the respiratory chain when cytosolic osmolarity is decreased. On isolated rat liver mitochondria incubated in hypoosmotic potassium chloride media, energetic parameters vary as in cells and oxidative phosphorylation efficiency is not affected. Cytosolic hyperosmolarity induced by sodium co-transported amino acids, per se, does not affect either matrix volume or energetic parameters. This is not the case in isolated rat liver mitochondria incubated in sucrose hyperosmotic medium. Indeed, in this medium, adenine nucleotide carrier is inhibited as the external osmolarity increases, which lowers the state 3 respiration close to state 4 level and consequently leads to a decrease in oxidative phosphorylation efficiency. When isolated rat liver mitochondria are incubated in KC1 hyperosmotic medium, state 3 respiratory rate, matrix volume and membrane electrical potential vary as a function of time. Indeed, matrix volume is recovered in hyperosmotic KC1 medium and this recovery is dependent on Pi-Kentry. State 3 respiratory rate increases and membrane electrical potential difference decreases during the first minutes of mitochondrial incubation until the attainment of the same value as in isoosmotic medium. This shows that matrix volume, flux and force are regulated as a function of time in KC1 hyperosmotic medium. Under steady state, neither matrix volume nor energetic parameters are affected. Moreover, NaCl hyperosmotic medium allows matrix volume recovery but induces a decrease in state 3 respiratory flux. This indicates that potassium is necessary for both matrix volume and flux recovery in isolated mitochondria. We conclude that hypoosmotic medium induces an increase in kinetic control both upstream and on the respiratory chain and changes the oxidative phosphorylation response to forces. At steady state, hyperosmolarity, per se, has no effect on oxidative phosphorylation in either isolated hepatocytes or isolated mitochondria incubated in KC1 medium. Therefore, potassium plays a key role in matrix volume, flux and force regulation. (Mol Cell Biochem 184: 107–121, 1998)
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Häussinger D, Lang F: Cell volume in the regulation of hepatic function: A mechanism for metabolic control. Biochim Biophys Acta 1071:331–350, 1991
Hallbrucker C, Vom Dahl S, Lang F, Häussinger D: Control of hepatic proteolysis by amino acids. The role of cell volume. Eur J Biochem 197:717–724, 1991
Bacquet A, Hue L, Meijer AJ, Van Woerkom GM, Plomb PJAM: Swelling of rat hepatocytes stimulates glycogen synthesis. J Biol Chem 265:955–959, 1990
Graf J, Haddad P, Häussinger D, Lang F: Cell volume regulation in liver. Renal Physiol Biochem 11:202–220, 1988
Häussinger D, Stehle T, Lang F: Volume regulation in liver: Further characterization by inhibitors and ionic substitution. Hepatology 11:243–254, 1990
Lang F, Stehle T, Häussinger D: Water, K+, H+, lactate and glucose fluxes during cell volume regulation in perfused rat liver. Pflüggers Arch 413:209–216, 1989
Häussinger D, Halbrucker C, Vom Dahl S, Lang F, Gerok W: Cell swelling inhibits proteolysis in perfused rat liver. Biochem. J 272:239–242, 1990
Häussinger D, Halbrucker C, Vom Dahl S, Decker S, Schweizer V, Lang F, Gerok W: Cell volume is a major determinant of proteolysis control in liver. FEBS Lett 283:70–72, 1991
Häussinger D, Lang F J: Exposure of perfused liver to hypotonic conditions modify cellular nitrogen metabolism. Cell Biochem 43:355–361, 1990
Hallbrucker C, vom Dahl S, Lang F, Haussinger D: Interactions between cell volume and hepatic nitrogen metabolism. Cont Nephrol 92:175–181, 1991
Bode B, Kilberg M: Amino acid dependent increase in hepatic system N activity is linked to cell swelling. J Biol Chem 266:7376–7381, 1991
Häussinger D, Lang F: The mutual interaction between cell volume and cell function: A new principle of metabolic regulation. Biochem CellBio 143:1–4, 1991
Meijer AJ, Bacquet A, Gustafson L, Van Woerkom GM, Hue L: Mechanism of activation of liver glycogen synthase by swelling. J Biol Chem 267:5823–5828, 1992
Bacquet A, Gaussin V, Bollen M, Stalmaus W, Hue L: Mechanism of activation of liver acetyl-CoA carboxylase by cell swelling. Eur J Biochem 217:1083–1089, 1993
Völkel H, Busch GL, Häussinger D, Lang F: Alkalinisation of acidic cellular compartments following cell swelling. FEBS Lett 338:27–30, 1994
Halestrap PA: The regulation of the matrix volume of mammalian mitochondria in vivo and in vitro and its role in the control of mitochondrial metabolism. Biochim Biophys Acta 973:355–382, 1989
Halestrap PA, Davidson MA, Potter WD: Mechanisms involved in the hormonal regulation of mitochondrial function through changes in the matrix volume. Biochim Biophys Acta 1018:278–281, 1990
Armston EA, Halestrap PA, Scott DR: The nature of the changes in liver mitochondrial function induced by glucagon treatment of rats. The effects of intramitochondrial volume, aging and benzyl alcohol. Biochim Biophys Acta 681:429–439, 1982
Otto AD, Ontko AJ: Structure-function relations between fatty acids oxidation and the mitochondrial inner membrane-matrix region. Eur J Biochem 129:479–485, 1982
Mathai CJ, Sauna EZ, John O, Sitaramam V: Rate limiting step in electron transport. Osmotically sensitive diffusion of quinones through voids in the bilayer. J Biol Chem 268:15442–15454, 1993
Nicholls DG, Lindberg O: Inhibited respiration and ATPase activity of rat liver mitochondria under conditions of matrix condensation. FEBS Lett 25:61–64, 1972
Sitaramam V, Rao NM: Oxidative phosphorylation in rat liver mito chondria: Influence of physical parameters. Ind J Biochem Biophys 28:401–407, 1991
Garlid KD: Mitochondrial cation transport: A progress report. J Bioenerg Biomembr 26:537–542, 1994
Garlid KD: On the mechanism of regulation of the mitochondrial K+H+ exchanger. J Biol Chem 255:11273–11279, 1980
Gunter TE: Transport of calcium by mitochondria. J Bioenerg Biomembr 26:465–469, 1994
Berry NM, Friend DS: High-yield preparation of isolated rat liver parenchymal cells: A biochemical and fine structural study. J Cell Biol 43:506–520, 1969
Groen AK, Sips HJ, Vervoorn RC, Tager JM: Intracellular compartmentation and control of alanine metabolism in rat liver parenchymal cells. Eur JBiochem 122:87–93, 1982
Krebs HA, Heiseleit K: Urea formation in the animal body. Hoppe-Seyler’s Z Physiol Chem 210:33–66, 1932
Quilan PT, Thomas AP, Armston AE, Halestrap AP: Measurement of the intramitochondrial volume in hepatocytes without cell disruption and its elevation by hormones and valinomycin. Biochem J 214:395–404, 1983
Nobes CD, Brand MD: A quantitative assessment of the use of 36C1’ distribution to measure plasma membrane potential in isolated hepatocytes. Biochim Biophys Acta 987:115–123, 1989
Nobes CD, Hay WW, Brand MD: The mechanism of stimulation of respiration by fatty acids in isolated hepatocytes. J Biol Chem 265:12910–12915, 1990
Scott ID, Nicholls DG: Energy transduction in intact synaptosomes. BiochemJ 186:21–33, 1980
Berry MN, Gregory RB, Grivell AR, Henly DC, Nobes CD, Phillips JW, Wallace PC: Intracellular mitochondrial membrane potential as an indicator of hepatocytes energy metabolism: Further evidence for thermodynamic control of metabolism. Biochim Biophys Acta 936:294–306, 1988
Hoek JB, Nicholls DG, Williamson JR: Determination of the mitochondrial protonmotive force in isolated hepatocytes. J Biol Chem 255:1458–1464, 1980
Murphy MP, Brand MD: The control of electron flux through cytochrome oxidase. Biochem J 243:499–505, 1987
Zuurendonck PF, Tager JM: Rapid separation of particulate components and soluble cytoplasm of isolated rat liver cells. Biochim Biophys Acta 333:393–399, 1974
Akerboom TPM, Van der Meer R, Tager JM: Techniques for the investigation of intracellular compartmentation. Tech Metab Res B 205:1–33, 1979
Krebs HA, Veech RL: In: S Papa, JM Tager, E Qualliariello, EC Slater (eds). Pyridine Nucleotide Interrelations. Adriatica Editrice, Bari, 1969, pp 329–382
Berenblum I, Chain E: Studies on the colorimetric determination of phosphate. Biochem J 32:286–294, 1938
Henrickson RL, Meredith SC: Amino acid analysis by reverse phase high performance liquid chromatography: Precolumn derivatization with phenylisothiocyanate. Anal Biochem 136:65–74, 1984
Cooper C, Lehninger AL: Oxidative phosphorylation by an enzyme complex from extracts of mitochondria. J Biol Chem 177:751–766, 1956
Gornall AG, Bardawill CJ, David MM: Determination of serum proteins by means ofthebiuret reaction. J Biol Chem 177:751–766, 1948
Rigoulet M, Devin A, Avéret N, Vandais B, Guérin B: Mechanisms of inhibition and uncoupling of respiration in isolated rat liver mito chondria by the general anesthetic 2,6-diisopropylphenol. Eur J Biochem 241:280–285, 1996
Rigoulet M, Guérin B: Phosphate transport and ATP-synthesis in yeast mitochondria: Effect of a new inhibitor: The tribenzyl phosphate. FEBS Lett 102:18–22, 1979
Rottenberg H.: The measurement of membrane potential and ApH in cells, organelles and vesicles. Meth Enzymol 55:547–569, 1979
Espié P, Guérin B, Rigoulet M: On isolated hepatocytes mitochondrial swelling induced in hypoosmotic medium does not affect the respiration rate. Biochim Biophys Acta 1230:139–146, 1995
Koretsky A, Balaban RS: Changes in pyridine nucleotides level alter oxygen consumption and extra-mitochondrial phosphate in isolated mitochondria: A31PNMR and NAD(P)H fluorescence study. Biochim Biophys Acta 893:398–408, 1987
Emaus RK, Grunwald R, Lemasters JJ: Rhodamine 123 as a probe of transmembrane potential in isolated rat liver mitochondria: Spectral and metabolic properties. Biochim Biophys Acta 850:436–448, 1986
Devin A, Guérin B, Rigoulet M: Dependence of flux size and efficiency of oxidative phosphorylation on external osmolarity in isolated rat liver mitochondria: Role of adenine nucleotide carrier. Biochim Biophys Acta 1273:13–20, 1996
Groen KA, Wanders AJR, Westerhoff VH, van der Meer R, Tager MJ: Quantification of the contribution of various steps to control mito chondrial respiration. J Biol Chem 257:2754–2757, 1982
Gellerich FN, Kunz WS, Bohnensack R: Estimation of flux control coefficient from inhibitor titration by non-linear regression. FEBS Lett 274:167–170, 1990
Groen AK: Quantification of control in studies on intermediary metabolism. Ph.D. Thesis, Amsterdam, 1984
Vignais PV, Vignais M, Defaye G: Adenosine diphosphate translocation in yeast mitochondria. Nature of the receptor site for carboxy-atractyloside. Biochemistry 12:1508–1519, 1973
Brown GC: Control of respiration and ATP synthesis in mammalians mitochondria and cells. Biochem J 284:1–13, 1992
Quentin E, Averet N, Guerin B, Rigoulet M: Temperature dependence of the coupling efficiency of rat liver oxidative phosphorylation: role of adenine nucleotide translocator. Biophys Biochem Res Com 202:816–821, 1994
Brierley GP, Baysal K, Jung DW: Cation transport systems in mitochondria: Na+ and K+ uniports and exchangers. J Bioenerg Biomembr 26:519–526, 1994
Beavis AD: On the inhibition of the mitochondrial inner membrane anion uniporter by cationic amphiphiles and others drugs. J Biol Chem 264:1508–1515, 1989
Sturgess NC, Ashford MLJ, Cook DL, Hales CN: The sulphonylurae receptor may be an ATP sensitive potassium channel. Lancet 8453:474–475, 1985
Inoue I, Nagase H, Kishi K, Higuti T: ATP-sensitive K+ channel in the mitochondrial inner membrane. Nature 352:244–247, 1991
Garlid KD, DiResta DJ, Beavis AD, Martin WH: On the mechanism by which dicyclohexylcarboxydiimide and quinine inhibits K+ transport in rat liver mitochondria. J Biol Chem 261:1529–1535, 1986
Halestrap AP, Quilan TP, Whipps ED, Armston EA: Regulation of the mitochondrial matrix volume in vivo and in vitro. The role of calcium. Biochem J 236:779–787, 1986
Halestrap AP, Dunlop LJ: Intramitochondrial regulation of fatty acids β-oxidation occurs between flavoproteins and ubiquinone. A role for changes in matrix volume. Biochem J 239:559–565, 1986
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1998 Springer Science+Business Media Dordrecht
About this chapter
Cite this chapter
Devin, A., Espié, P., Guérin, B., Rigoulet, M. (1998). Energetics of swelling in isolated hepatocytes: A comprehensive study. In: Saks, V.A., Ventura-Clapier, R., Leverve, X., Rossi, A., Rigoulet, M. (eds) Bioenergetics of the Cell: Quantitative Aspects. Developments in Molecular and Cellular Biochemistry, vol 25. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-5653-4_9
Download citation
DOI: https://doi.org/10.1007/978-1-4615-5653-4_9
Publisher Name: Springer, Boston, MA
Print ISBN: 978-1-4613-7587-6
Online ISBN: 978-1-4615-5653-4
eBook Packages: Springer Book Archive