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Non-linear release of Bohr protons with haemoglobin-oxygenation in the blood of two teleost fishes; carp (Cyprinus carpio) and tambaqui (Colossoma macropomum)

Abstract

Some teleost fishes exhibit a non-linear release of H+ with haemoglobin oxygen saturation (SO2) in whole blood that may be related to the Root effect and a low apparent cooperativity in oxygen binding. To further investigate this correlation, the relationship between red cell pH (pHi) and SO2 was evaluated in two teleost fishes, the carp (Cyprinus carpio) and tambaqui (Colossoma macropomum). Carp possess a relatively small Root effect within the physiological pH range, while that in tambaqui is relatively large. It was therefore hypothesized that both species would possess a non-linear release of H+ with SO2 under in vivo conditions, but that the degree of non-linearity would be less pronounced in carp than in tambaqui. Red cell suspensions of tambaqui showed a marked non-linear relationship between pHi and SO2 at in vivo pH values (pHe=7.66), where the majority of Bohr protons were released between 50 and 100% SO2. In whole blood of carp, the relationship between pHi and SO2 was almost linear at normal resting extracellular pH values (pHe=8.10) where Hills n 50 (cooperativity) was 1.3. Under acidic conditions (pHe=7.11), n 50 decreased to 0.9 and the release of H+ with SO2 became slightly non-linear. The reduction in pH in carp blood was associated with the onset of the Root effect, and the oxygen tension at half saturation (P50) increased from 3.8 to 38.2 mmHg. The relationship between total CO2 and SO2 in carp whole blood changed from being almost linear at pHe 8.1 to being non-linear at pHe 7.1, consistent with the relationship between pHi and SO2. Thus, possession of a Root effect could be a prerequisite for a non-linear release of Bohr protons with oxygenation, but the expression in whole blood may depend upon a species-specific cooperativity threshold. The non-linear release of Bohr protons with oxygenation may be a general phenomenon in teleosts with important implications for gas transport and acid-base homeostasis.

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References

  • Benesch, R.E., Benesch, R. and Yung, S. 1973. Equations for the spectrophotometric analysis of hemolgobin mixtures. Anal. Biochem. 55: 245–248.

    Google Scholar 

  • Bourne, P.K. and Cossins, A.R. 1982. On the instability of K+ in-flux in erythrocytes of the rainbow trout, Salmon gairdneri, and the role of catecholamine hormones in maintaining in vivo influx activity. J Exp Biol. 101: 93–104.

    Google Scholar 

  • Brauner, C.J. and Jensen, F.B. 1999. O2 and CO2 exchange in fish: The non-linear release of Bohr/Haldane protons with oxygenation. In: Biology of Tropical Fishes. pp. 394–400. Edited by A.L. Val and V.M.F. Almeida-Val, INPA, Manaus.

  • Brauner, C.J. and Randall, D.J. 1996. The interaction between oxygen and carbon dioxide movements in fishes. Comp. Biochem. Physiol. 113A: 83–90.

    Google Scholar 

  • Brauner, C.J. and Randall, D.J. 1998. The linkage between oxygen and carbon dioxide transport. In: Fish Physiology; Fish Respiration. pp. 283–319. Edited by S.F. Perry and B. Tufts. Academic Press, New York.

    Google Scholar 

  • Brauner, C.J. and Weber, R.E. 1998. Hydrogen ion titrations of the anodic and cathodic haemoglobin components of the European eel Anguilla anguilla. J. exp. Biol. 201: 2507–2514.

    Google Scholar 

  • Brauner, C.J., Gilmour, K.M. and Perry, S.F. 1996. Effect of haemoglobin oxygenation on Bohr proton release and CO2 excretion in the rainbow trout. Respir. Physiol. 106: 65–70.

    Google Scholar 

  • Brauner, C.J., Thorarensen, H., Gallaugher, P., Farrell, A.P. and Randall, D.J. 2000. The interaction between O2 and CO2 exchange in rainbow trout during graded sustained exercise. Resp Physiol. 119(1): 83–96.

    Google Scholar 

  • Brittain, T. 1987. The Root effect. Comp. Biochem. Physiol. 86B: 473–481.

    Google Scholar 

  • Cameron, J.N. 1971. Rapid method for determination of total carbon dioxide in small blood samples. J. Appl. Physiol. 31: 632–634.

    Google Scholar 

  • Chien, J.C.W. and Mayo, K.H. 1980. Carp hemoglobin. II. The alkaline Bohr effect. J. Biol. Chem. 255: 9800–9806.

    Google Scholar 

  • Christoforides, C. and Hedley-Whyte, J. 1969. Effect of temperature and hemoglobin concentration on solubility O2 in blood. J. Appl. Physiol. 27(5): 592–596.

    Google Scholar 

  • Costa, O.T.F. (1995) Efeito do pH da água sobre os parâmetros hematológicos, eletrólitos e equilíbrio ácido base dop sangue de Colossoma macropomum (Characiformes, Serrasalmidae). MSc dissertation. INPA, Manaus, AM. 108 pp.

  • Heisler, N. 1986. Parameters and methods in acid-base physiology. In: Techniques in comparative respiratory physiology; an experimental approach. pp. 305–332. Edited by C.R. Bridges and P.J. Butler. Cambridge University Press, Cambridge.

    Google Scholar 

  • Ikeda-Saito, M., Yonetani, T. and Gibson, Q.H. 1983. Oxygen equilibrium studies on hemoglobin from the bluefin tuna (Thunnus thynnus). J. Mol. Biol. 168: 673–686.

    Google Scholar 

  • Imai, K. and Yonetani, T. 1975. pH dependence of the Adair constants of human hemoglobin: Nonuniform contribution of successive oxygen bindings to the alkaline Bohr effect. J. Biol. Chem. 250: 2227–2231.

    Google Scholar 

  • Jensen, F.B. 1986. Pronounced influence of Hb-O2 saturation on red cell pH in tench blood in vivo and in vitro. J. Exp. Zool. 238: 119–124.

    Google Scholar 

  • Jensen, F.B. 1989. Hydrogen ion equilibria in fish haemoglobins. J. exp. Biol. 143: 225–234.

    Google Scholar 

  • Jensen, F.B. 1990. Nitrite and red cell function in carp: control factors for nitrite entry, membrane potassium ion permeation, oxygen affinity and methaemoglobin formation. J. Exp. Biol. 152: 149–166.

    Google Scholar 

  • Jensen, F.B. 1991. Multiple strategies in oxygen and carbon dioxide transport by haemoglobin. In: Physiological Strategies for Gas Exchange and Metabolism. pp. 55–78. Edited by A.J. Woakes, M.K. Greishaber and C.R. Bridges. Cambridge University Press, Cambridge.

    Google Scholar 

  • Jensen, F.B. 1992. Influence of haemoglobin conformation, nitrite and eicosanoids on K+ transport across the carp red blood cell membrane. J. Exp. Biol. 171: 349–371.

    Google Scholar 

  • Jensen, F.B. and Weber, R.E. 1982. Respiratory properties of tench blood and hemoglobin. Adaptation to hypoxic-hypercapnic water. Mol. Physiol. 2: 235–250.

    Google Scholar 

  • Lowe, T.E., Brill, R.W. and Cousins, K.L. 1998. Responses of the red blood cells from two high-energy-demand teleosts, yellowfin tuna (Thunnus albacares) and skipjack tuna (Katsuwonus pelamis), to catecholamines. J. Comp. Physiol. B 168: 405–418.

    Google Scholar 

  • Narahara, A., Bergman,6H.L., Laurent, P., Maina, J.N., Walsh, P.J. and Wood, C.M. 1996. Respiratory physiology of the Lake Magadi tilapia (Oreochromis alcalicus grahami), a fish adapted to a hot, alkaline, and frequently hypoxic environment. Physiol. Zool. 69(5): 1114–1136.

    Google Scholar 

  • Pelster, B. and Weber, R.E. 1990. Influence of organic phosphates on the Root effect of multiple fish haemoglobins. J. Exp. Biol. 149: 425–437.

    Google Scholar 

  • Soivio, A., Nyholm, K. and Westman, K. 1975. A technique for repeated sampling of the blood of individual resting fish. J. Exp. Biol. 62: 207–217.

    Google Scholar 

  • Tetens, V. and Lykkeboe, G. 1981. Blood respiratory properties of rainbow trout, Salmo gairdneri: Responses to hypoxia acclimation and anoxic incubation of blood in vitro. J. Comp. Physiol. 134: 117–125.

    Google Scholar 

  • Tucker, V.A. 1967. A method for oxygen content and dissociation curves on microliter blood samples.J. Appl. Physiol. 23(3): 407–410.

    Google Scholar 

  • Val, A.L. 1995. Oxygen transfer in fish: morphological and molecular adjustments. Braz. J. Med. Biol. Res. 28: 1119–1127.

    Google Scholar 

  • Val, A.L. and Almeida-Val, V.M.F. 1995. Fishes of the Amazon and Their Environment: Physiological and Biochemical Aspects. Springer-Verlag, Berlin.

    Google Scholar 

  • Val, A.L., Mazur, C.F., De Salvo-Souza, R.H. and Iwama, G.K. 1994. Effects of experimental anaemia on intra-erythrocytic phosphate levels in rainbow trout, Oncorhynchus mykiss. J. Fish Biol. 45: 269–277.

    Google Scholar 

  • Val, A.L., DeMenzes, G.C., and Wood, C.M. 1998. Red blood cell adrenergic responses in Amazonian teleosts. J. Fish Biol. 52: 83–93.

    Google Scholar 

  • Walsh, P.J., Wood, C.M. and Moon, T.W. 1998. Red blood cell metabolism. In: Fish Physiology; Fish Respiration. pp. 41–73. Edited by S.F. Perry and B. Tufts Academic Press, New York.

    Google Scholar 

  • Weber, R.E., Jensen, F.B. and Cox, R.P. 1987. Analysis of teleost hemoglobin by Adair and Monod-Wyman-Changeux models. Effects of nucleotide triphosphates and pH on oxygenation of tench hemoglobin. J. Comp. Physiol. B 157: 145–152.

    Google Scholar 

  • Weber, R.E. and Lykkeboe, G. 1978. Respiratory adaptations in carp blood. Influences of hypoxia, red cell organic phosphates, divalent cations and CO2 on hemoglobin-oxygen affinity. J. Comp. Physiol. 128: 127–137.

    Google Scholar 

  • Wells, R.M.G. and Weber, R.E. 1989. The measurement of oxygen affinity in blood and haemoglobin solutions. In: Techniques in comparative respiratory physiology; an experimental approach. pp. 279–303. Edited by C.R. Bridges and P.J. Butler. Cambridge, University Press.

  • Wood, C.M., Wilson, R.W., Gonzalez, R.J., Patrick, M.L., Bergman, H.L., Narahara, A., and Val, A.L. 1998. Responses of an Amazonian teleost, the tambaqui (Colossoma macropomum), to low pH in extremely soft water. Physiol. Zool. 71(6): 658–670.

    Google Scholar 

  • Wolf, K. 1963. Physiological salines for fresh-water teleosts. Prog. Fish. Cult. 25: 135–140.

    Google Scholar 

  • Wyman, 1964. Linked functions and reciprocal effects in haemoglobin: A second look. Adv. Protein Chem. 19: 195–205.

    Google Scholar 

  • Zeidler, R. and Kim, D.H. 1977. Preferential hemolysis of postnatal calf red cells induced by internal alkalinization.J. Gen. Physiol. 70: 385–401.

    Google Scholar 

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Brauner, C., Wang, T., Val, A. et al. Non-linear release of Bohr protons with haemoglobin-oxygenation in the blood of two teleost fishes; carp (Cyprinus carpio) and tambaqui (Colossoma macropomum). Fish Physiology and Biochemistry 24, 97–104 (2001). https://doi.org/10.1023/A:1011944407938

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  • DOI: https://doi.org/10.1023/A:1011944407938

  • Bohr effect
  • Colossoma macropomum
  • Cyprinus carpio
  • haemoglobin
  • Haldane effect
  • Root effect
  • red blood cell pH