Skip to main content
SpringerLink
Log in

Identification and immunocytochemical localization of two different carbonic anhydrase isoenzymes in teleostean fish erythrocytes and gill epithelia

  • Published:
Histochemistry Aims and scope Submit manuscript

Summary

Carbonic anhydrase was purified from the gills (CAB) of the rainbow trout Salmo gairdneri and from erythrocytes (CAE) of the fresh water carp Cyprinus carpio. The purification of the isozymes was confirmed by SDS acrylamide gel electrophoresis. Antibodies against the purified CAB and CAE were then raised in rabbits. Specificity was verified by immunoblotting. No cross-reaction was found between them, using the immunodot technique. CAB antiserum was used to specifically localize gill CA in the trout. Immunoperoxidase labelling revealed a concentration of enzyme on the apical region of the outer layer of the gill epithelial cells. The inner layer of the epithelium was only weakly positive. Results obtained using the immuno-gold technique confirmed the immunoperoxidase labelling: there was a concentration of label in the apical regions of chloride cells. In mucous cells, only the mucous granules were labelled. In the lamellae, the label was distributed in the apical part of the pavement cells. The villi and microplicae were strongly positive. CAE antiserum stained the red blood cells. The discrepancy between histochemical localization in the gill or in the opercular skin of killifish and our present immunolocalization was discussed.

It was concluded that the most typical localization of CA is on the apical surface of the lamellar epithelium lying in contact with the environment. The result suggests that one of the main roles of gill CA may be to facilitate the diffusion of CO2 from blood to water.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Altman LG, Schneider BG, Papermaster DS (1984) Rapid embedding of tissues in Lowicryl k4M for immunoelectron microscopy. J Histochem Cytochem 32:1217–1223

    Google Scholar 

  • Brown D, Kumpulainen T, Roth J, Orci L (1983) Immunohistochemical localization of carbonic anhydrase in postnatal and adult rat kidney. Am J Physiol 245:F110-F118

    Google Scholar 

  • Brown D, Garcia-Segura LM, Orci L (1984) Carbonic anhydrase is present in olfactory receptor cells. Histochemistry 80:307–309

    Google Scholar 

  • Burnett LE, McMahon BR (1985) Facilitation of CO2 excretion by carbonic anhydrase located on the surface of the basal membrane of crab gill epithelium. Respir Physiol 62:344–348

    Google Scholar 

  • Cameron JN (1979) Excretion of CO2 in water breathing animals. A short review. Mar Biol Lett 1:3–13

    Google Scholar 

  • Churg A (1973) Carbonic anhydrase histochemistry: evidence for non-enzymatic reaction and artifact production. Histochemie 36:293–302

    Google Scholar 

  • Delaunoy JP, Roussel G, Mandel P (1977) Localisation immunohistochimique de l'anhydrase carbonique C dans le système nerveux central du rat. C R Acad Sci Ser D 285:801–803

    Google Scholar 

  • De Renzis G, Maetz J (1973) Studies on the mechanism of chloride absorption by the gold fish gill: relation with acid-base regulation. J Exp Biol 59:339–358

    Google Scholar 

  • Dimberg K, Hoglund LB, Knutsson PG, Ridderstrale Y (1981) Histochemical localization of carbonic anhydrase in gill lamella from young salmon (Salmo salar L.) adapted to fresh and salt water. Acta Physiol Scand 112:218–220

    Google Scholar 

  • Evans DH (1982) Salt and water exchange across vertebrate gill. In: Houlihan DF, Rankin JC, Shuttelworth TJ (eds) Gill. Cambridge University Press, Cambridge, pp 149–172

    Google Scholar 

  • Evans DH, Cameron NH (1986) Gill ammonia transport. J Exp Zool 239:17–23

    Google Scholar 

  • Filippi D, Sciaky M, Limozin N, Laurent G (1978) Anhydrase carbonique du système nerveux central du rat. Isolement et propriétés. Biochimie 60:99–102

    Google Scholar 

  • Gardaire E, Avella M, Isaia J, Bornancin M, Mayer-Gostan N (1985) Estimation of sodium uptake through the gill of the rainbow trout, Salmo gairdneri. J Exp Biol 44:181–189

    Google Scholar 

  • Girard JP, Istin M (1975) Iosenzymes de l'anhydrase carbonique d'un poisson enryphaline, variation en relation avec l'osmorégulation. Biochim Biophys Acta 381:221–232

    Google Scholar 

  • Hall GE, Schraer R (1983) Characterization of a high activity carbonic anhydrase isozyme purified from erythrocytes of Salmo gairdneri. Comp Biochem Physiol 75B1:81–92

    Google Scholar 

  • Hansson HPJ (1967) Histochemical demonstration of carbonic anhydrase activity. Histochemie 11:112–128

    Google Scholar 

  • Haswell MS, Randall DJ (1978) The pattern of carbon dioxide excretion in the rainbow trout salmo gairdneri. J Exp Biol 72:17–24

    Google Scholar 

  • Haswell MS, Randall DJ, Perry SF (1980) Fish gill carbonic anhydrase: acid-base or salt transport. Am J Physiol 238:R240-R244

    Google Scholar 

  • Heisler N (1984) Acid-base regulation in fishes. In: Hoar WS, Randall DJ (eds) Fish physiology, vol 10a. Academic Press, New York, pp 315–401

    Google Scholar 

  • Henry RP, Dodgson SJ, Forster RE, Storey BT (1986) Rat lung carbonic anhydrase: activity, localization and isozymes. J Appl Physiol 66(2):638–645

    Google Scholar 

  • Kumpulainen T (1979) Immunohistochemical localization of human carbonic anhydrase isozyme C. Histochemistry 62:271–280

    Google Scholar 

  • Kumpulainen T (1981) Human carbonic anhydrase isozyme C. Effects of some fixative on the antigenicity and improvements in the method of localization. Histochemistry 72:425–431

    Google Scholar 

  • Lacy ER (1983) Histochemical and biochemical studies of carbonic anhydrase activity in the opercular epithelium of the euryhaline teleost, fundulus heteroclitus. Am J Anat 166:19–39

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Google Scholar 

  • Laurent P (1984) Gill internal morphology. In: Hoar WS, Randall DJ (eds) Fish physiology, vol 10a. Academic Press, New York, pp 73–183

    Google Scholar 

  • Laurent P (1988) Gill structure and function: fish. In: Wood S (ed) Comparative pulmonary physiology: Current concepts. Marcel Dekker, New York, pp 69–120

    Google Scholar 

  • Laurent P, Dunel S (1980) Morphology of gill epithelia in fish. Am J Physiol 238:R147–159

    Google Scholar 

  • Laurent P, Dunel S, Barets A (1969) Localisation histochimique de l'anhydrase carbonique au niveau des chémorécepteurs artériels de mammifères, des batraciens et des poissons. Histochemie 17:99–108

    Google Scholar 

  • Laurent P, Hobe H, Dunel-Erb S (1985) The role of environmental sodium chloride relative to calcium in gill morphology of fresh water salmonid fish. Cell Tissue Res 240:675–692

    Google Scholar 

  • Lauweryns JM, Ranst LV, Verhofstad AAJ (1986) Ultrastructural localization of serotonin in the intrapulmonary neuroepithelial bodies of neonatal rabbits by use of immunoelectron microscopy. Cell Tissue Res 243:455–459

    Google Scholar 

  • Leong MML, Hilstein C, Pannell R (1986) Luminescent detection method for immunodot, Western, and Southern blots. J Histochem Cytochem 34:1645–1650

    Google Scholar 

  • Maetz J, Garcia Romeu F (1964) The mechanism of sodium and chloride uptake by the gills of a fresh water fish Carassius auratus. II. Evidence for NH4/Na+ and HCO3/Cl− exchanges. J Genet Physiol 47:1209–1227

    Google Scholar 

  • Maren TH (1967) Carbonic anhydrase: chemistry, physiology and inhibition. Physiol Rev 47:595–781

    Google Scholar 

  • Marriq C, Giraud N, Laurent G (1974) Isolement de l'anhydrase carbonique erythrocytaire bovine par chromatographie d'affinité. C R Acad Sci 279:1325–1328

    Google Scholar 

  • Muther TF (1977) On the lack of specificity of the cobalt bicarbonate method for carbonic anhydrase. J Histochem Cytochem 25:1043–1050

    Google Scholar 

  • Perry SF, Dayse PS, Daxboeck C, Randall DJ (1982) A comparison of CO2 excretion in a spontaneously ventilating blood-perfused trout preparation saline-perfused gill preparation: contribution of the branchial epithelium and red blood cells. J Exp Biol 101:47–60

    Google Scholar 

  • Piiper J (1969) Rates of chloride-bicarbonate exchange between red cells and plasma. In: Forester RE et al (ed) CO2: Chemical, biochemical and physiological aspects. Nasa SP-188, Washington, pp 267–273

  • Randall DJ, Daxboeck C (1984) Oxygen and carbon dioxide transfer across fish gill. In: Hoar WS, Randall DJ (eds) Fish physiology. Academic Press, New York, pp 263–314

    Google Scholar 

  • Rickli EE, Ghazanfar SAS, Gibbons BH, Edsall GT (1964) Carbonic anhydrase from human erythrocytes. J Biol Chem 239:1065–1078

    Google Scholar 

  • Spicer SS, Sens MA, Tashian RE (1982) Immunocytochemical demonstration of carbonic anhydrase in human epithelial cells. J Histochem Cytochem 30:864–873

    Google Scholar 

  • Swenson ER (1979) Teleost erythrocyte carbonic anhydrase: isolation and partial characterization. Bull M Desert Isl Biol Lab 19:74–77

    Google Scholar 

  • Thurston RV, Russo RC, Vinogradov GA (1981) Ammonia toxicity to fishes. Effect of pH on the toxicity at the un-ionized ammonia species. Environ Sci Technol 15(7):837–840

    Google Scholar 

  • Towbin H, Staekelin T, Gordon T (1979) Electrophoretic transfer of protein from polyacrylamide gels to nitrocellulose sheats: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354

    Google Scholar 

  • Wilbur KM, Andersson NG (1948) Electrometric and calorimetric determination of carbonic anhydrase. J Biol Chem 176:147–154

    Google Scholar 

  • Wright P, Heming T, Randall D (1986) Downstream pH changes in water flowing over the gills of rainbow trout. J Exp Biol 126:499–512

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Laboratoire de Morphologie Fonctionnelle et Ultrastructurale des Adaptations, CNRS, B.P. 20CR, F-67037, Strasbourg Cedex, France

    S. M. Rahim & P. Laurent

  2. Centre de Neurochimie, CNRS, Strasbourg, France

    J. -P. Delaunoy

Authors
  1. S. M. Rahim
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J. -P. Delaunoy
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. P. Laurent
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Rahim, S.M., Delaunoy, J.P. & Laurent, P. Identification and immunocytochemical localization of two different carbonic anhydrase isoenzymes in teleostean fish erythrocytes and gill epithelia. Histochemistry 89, 451–459 (1988). https://doi.org/10.1007/BF00492602

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00492602

Keywords

Search

Navigation