Abstract
Temperature-induced alterations in the activity of the enzyme lactate dehydrogenase (LDH) were evaluated in the freshwater prawn Macrobrachium malcolmsonii in three groups of juveniles: controls maintained at 27±2 °C; test prawns exposed to 35 °C; test prawns exposed to 15 °C. Changes in LDH activity and lactate levels in key tissues were assessed after 48 hrs. LDH in the skeletal muscle of the prawns was also subjected to kinetic analysis at different temperatures. Native polyacrylamide gel electrophoresis (PAGE) analysis and colorimetric estimation revealed decreased LDH activity (compared to controls) in the gill, heart and haemolymph, but not in the skeletal muscle or hepatopancreas, of test prawns exposed to 15 °C; however, lactate levels were significantly lower in all the tissues of these test prawns. Conversely, prawns exposed to 35 °C revealed elevated LDH activity in all the tissues, barring the skeletal muscle, while lactate levels were significantly higher (compared to controls) in all the tissues of these prawns. Kinetic analysis of LDH in the skeletal muscle at different assay temperatures revealed temperature-dependent kinetic properties. The differences observed in LDH activity and levels of lactate in various tissues of prawns exposed to low and high temperatures suggest aerobic and anaerobic patterns of pyruvate metabolism at respective temperatures. The results obtained by kinetic analysis of LDH in the skeletal muscle suggest the occurrence of an adaptative response involving this enzyme that enables M. malcolmsonii to cope with effects of thermal stress.
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References
Bakun, A. 1996. Patterns in the Ocean Processes and Marine Population Dynamics. California Sea Grant College System, La Jolla. pp 323.
Beamish, R.J. 1995. Climate change and northern fish populations. Canadian Special Publication of Fisheries and Aquatic Sciences. 121: pp. 739.
Bergmeyer, H.U. and Bernt, E. 1974. In: Methods of Enzymatic Analysis. Vol. 2, pp. 574–593. Edited by H.U. Bergmeyer. Academic Press, Inc, USA.
Brand, M.D. 1990. The contribution of the leak of protons across the mitochondrial inner membrane to standard metabolic rate. J. Ther. Biol. 145: 267–286.
Brand, M.D., Chien, L.F., Ainscow, E.K. and Rolfe, R.K. 1994. The causes and functions of mitochondrial proton leak. Biochem. Biophys. Acta. 1187: 132–139.
Chernyi, V.G., Chizhevich, E.P. and Shukoliukov, S.A. 1976. Lactate dehydrogenase isoenzymes in the eye, cardiac and skeletal muscles of several decapods. Zh. Evol. Biokhim. Fiziol. 12(6): 521–6.
Danilenko, A.N., Persikov, A.V., Polosukhina, E.S., Klyachko, O.S., Esipova, N.G. and Ozernyuk, N.D. 1998. Thermodynamic properties of lactate dehydrogenase from muscles of fishes adapted to different environmental temperatures. Biofizika. 43: 26–30.
Dietze, A.A., Lubrano, A. and Rabinstein, H.M. 1970. Disc electrophoresis of lactate dehydrogenase isoenzymes. Clin. Chem. Acta. 27: 225–230.
Eszes, C.M., Sessions, R.B., Clarke, A.R., Moreton, K.M. and Holbrook, J.J. 1996. Removal of substrate inhibition in a lactate dehydrogenase from human muscle by a single residue change. FEBS Lett. 399, 193–197.
Fields, P.A. and Somero, G.N. 1998. Hot spots in cold adaptation: localized increases in conformational flexibility in lactate dehydrogenase A4 orthologs of Antarctic notothenioid fishes. Proc. Natl. Acad. Sci. USA. 95: 11476–11481.
Fields, P.A., Kim, Y.S., Carpenter, J.F and Somero, G.N. 2002. Temperature adaptation in Gillichthys (Teleost: Gobiidae) A(4)-lactate dehydrogenases: identical primary structures produce subtly different conformations. J. Exp. Biol. 205: 1293–1303.
Fields, P.A., Wahlstrand, B.D. and Somero, G.N. 2001. Intrinsic versus extrinsic stabilization of enzymes. The interaction of solutes and temperature on A4-lactate dehydrogenase orthologs from warm-adapted and cold-adapted marine fishes. Eur. J. Biochem. 268: 4497–4505.
Fine, I.H. and Costello, L.A. 1963. In: Methods in Enzymology. Vol VI, pp. 958–972. Academic Press, New York and London.
Frederich, M. and Portner, H.O. 2000. Oxygen limitation of thermal tolerance defined by cardiac and ventilatory performance in the spider crab, Maja squinado. Am. J. Physiol. Regulatory Integrative Comp Physiol. 279: 1531–1538.
Gutmann, I. and Wahlefeld, A.U. 1974. In: Methods of Enzymatic Analysis. Vol. 3, pp. 1464–1468. Edited by H.U. Bergmeyer. Academic Press, Inc, USA.
Hewitt, C.O., Eszes, C.M., Sessions, R.B., Moreton, K.M., Dafforn, T.R., Takei, J., Dempsey, C.E., Clarke, A.R and Holbrook, J.J. 1999. A general method for relieving substrate inhibition in lactate dehydrogenases. Protein. Eng. 12, 491–496.
King, J. 1965. Practical Clinical Enzymology. Van Nostrand Co Ltd, London.
Kliachkoos., Polosukhina, E.S. and Ozerniuk, N.D. 1993. Temperature causes structural and functional changes in lactate dehydrogenase from fish skeletal muscles. Biofizika. 38(4): 596–601.
Koban, M. 1986. Can cultured teleost hepatocytes show temperature acclimation?. Am. J. Physiol. 250(2 pt 2): 211–220.
Lannig, G., Eckerle, L., Serendero, I., Sartoris, F.J., Fischer, T., Knust, R., Johansen, T., and Portner, H.O. 2003. Temperature adaptation in euthermal cod (Gadus morhua): comparison of mitochondrial enzyme capacities in boreal and arctic populations. Mar. Biol. 142: 589–599.
Lowry, O.H., Rosebrough, N.J., Farr, A.L., and Randall, R.J. 1951. Protein measurement with folin phenol reagent. J. Biol. Chem. 193: 265–275.
Mauro, N.A. and Mangum, C.P. 1982. The role of the blood in the temperature dependence of oxidative metabolism in decapod crustaceans. Intraspecific responses to seasonal differences in temperature. J. Exp. Zool. 219: 179–188.
Panepucci, L., Fernandes, M.N., Sanches, J.R. and Rantin, F.T. 2000. Changes in lactate dehydrogenase and malate dehydrogenase activities during hypoxia and after temperature acclimation in the armored fish, Rhinelepis strigosa(Siluriformes, Loricariidae). Rev. Bras. Biol. 60(2): 353–60.
Persikov, A.V., Danilenko, A.N., Klyachko, O.S. and Ozernyuk, N.D. 1999. A comparative study of conformational stability of lactate dehydrogenase from loach skeletal muscles, adapted to different temperatures, using differential scanning microcalorimetry. Biofizika. 44: 32–37.
Pierce, V.A. and Crawford, D.L. 1997. Phylogenetic analysis of thermal acclimation of the glycolytic enzymes in the genus Fundulus. Physiol Zool. 70(6): 597–609.
Place, A.R. and Powers, D.A. 1984. Kinetic characterization of the lactate dehydrogenase(LDH-B4) allozymes of Fundulus heteroclitus. J. Biol. Chem. 259(2): 1309–1318.
Portner, H.O. 2001. Climate change and temperature-dependent biogeography: Oxygen limitation of thermal tolerance in animals. Naturwissenschaften 88: 137–146.
Portner, H.O., Hardewig, I., Sartoris, F.J. and Van Dijk, P.L.M. 1998. In: Cold Ocean Physiology. pp. 88–120. Edited by H.O. Portner and R. Playle. Cambridge University Press, UK.
Sharpe, M., Love, C and Marshall, C. 2001. Lactate dehydrogenase from the Antarctic eelpout, Lycodichthys dearborni. Ploar. Biol. 24: 258–269.
Somero, G.N. 1973. Thermal modulation of pyruvate metabolism in the fish Gillichthys mirabilis:the role of lactate dehydrogenase. Comp. Biochem. Physiol. 44B: 205–209.
Sommer, A., and Portner, H.O. 1999. Exposure of Arenicola marina(L) to extreme temperatures: adaptive flexibility of a boreal and a subpolar population. Mar. Ecol. Prog. Ser. 181: 215–226.
Sommer, A., Klein B. and Portner, H.O. 1997. Temperature induced anaerobiosis in two populations of the polychaete worm Arenicola marina(L). J. Comp. Physiol (B). 167: 25–35.
Vander Helm. 1961. Simple method of demonstrating lactate dehydrogenase isozymes. Lancet 11: pp. 108.
Wieme. 1974. In: Methods of Enzymatic Analysis. Vol. 2, 593–602. Edited by H.U. Bergmeyer. Academic Press, Inc, USA.
Zakhartsev, M., Johansen, T., Portner, H.O and Blust, R. 2004. Effects of temperature acclimation on lactate dehydrogenase of cod (Gadus morhua): genetic, kinetic and thermodynamic aspects. J. Exp. Biol. 207: 95–112.
Zar, J.H. 1984. In Biostatistical analysis. Edited by Kurtz. 2nd Edition, pp. 185–197. Prentice Hall Int. Inc., New Jersey.
Zielinski, S. and Portner, H.O. 1996. Energy metabolism and ATP free-energy change of the intertidal worm, Sipunculus nudus, below a critical temperature. J. Comp. Physiol(B). 166: 492–500.
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Selvakumar, S., Geraldine, P. Thermal modulation of pyruvate metabolism in the freshwater prawn Macrobrachium malcolmsonii: the role of lactate dehydrogenase. Fish Physiology and Biochemistry 29, 149–157 (2003). https://doi.org/10.1023/B:FISH.0000035935.33689.ea
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DOI: https://doi.org/10.1023/B:FISH.0000035935.33689.ea
- Crustaceans
- enzyme kinetics
- LDH
- Macrobrachium malcolmsonii
- prawn
- pyruvate metabolism
- temperature stress