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Transport adenosine triphosphatase activity in the rat cornea

A comparative histochemical study

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Summary

The sodium-potassium activated adenosine triphosphatase (NaKATPase) activity of the rat cornea was investigated histochemically using a Pb2+-precipitation technique in which adenosine triphosphate (ATP) is used as substrate and two methods for potassium-dependent para-nitrophenyl-phosphatase (K-NPPase) activity.

With all the three techniques used it was demonstrated that the sodium-potassium-activated adenosine triphosphatase (NaK-ATPase) activity is localized in the cell membranes of the endothelium whereas a much weaker activity was observed in the epithelium. When the Pb2+-technique was used, the epithelial cell membranes showed a weaker reaction in the presence of ouabain. This activity was only Mg2+-dependent and was presumably due to an Mg2+-dependent ATPase.

The validity of the histochemical techniques for NaK-ATPase activity is discussed. The results emphasize the importance of the endothelium as the main site of Na+ transport in the cornea. Small amounts of the enzyme are also present in the epithelium, which seems to be rich in Mg2+-ATPase. Provided that careful controls are performed, all the methods give consistent results in the cornea.

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References

  • Albers, R.W.:Biochemical aspects of active transport. Ann. Rev. Biochem. 36, 727–757 (1967)

    Google Scholar 

  • Albers, R.W., Koval, G.J.: Sodium-potassium-activated adenosine triphosphatase VII. Concurrent inhibition of Na+-K+-adenosine triphosphatase and activation of K+-nitrophenylphosphatase activities. J. biol. Chem, 247, 3088–3092 (1972)

    Google Scholar 

  • Anderson, E.I.: Personal communication to J. Fischbarg. Acknowledged in Fischbarg and Lim (1974)

  • Anderson, E.I., Fischbarg, J., Spector, A.: Disulfide stimulation of fluid transport and effect on ATP level in rabbit corneal endothelium. Exp. Eye Res. 19, 1–10 (1974)

    Google Scholar 

  • Baum, J.L.: Nucleoside phosphatase staining of corneal stromal nerves. Arch. Ophthal., 82, 356–360 (1969)

    Google Scholar 

  • Beeuwkes, R., Rosen, S.: Renal sodium-potassium adenosine triphosphatase. Optical localization and x-ray microanalysis. J. Histochem. Cytochem. 23, 828–839 (1975)

    Google Scholar 

  • Bonting, S.L.: Sodium-potassium activated adenosine triphosphatase and cation transport. Membranes and ion transport. Vol. 1, Chap. 8, p. 257, New York: John Wiley and Sons, Inc. 1970

    Google Scholar 

  • Bonting, S.L., Simon, K.A., Hawkins, N.M.: Studies on sodium-potassium-activated adenosine triphosphatase. I Quantitative distribution in several tissues of the cat. Arch. Biochem. Biophys. 95, 416–423 (1961)

    Google Scholar 

  • Borgers, M.: The cytochemical application of new potent inhibitors of alkaline phosphatases. J. Histochem. Cytochem. 21, 812–824 (1973)

    Google Scholar 

  • Dikstein, S., Maurice, D.M.: The metabolic basis to the fluid pump in the cornea. J. Physiol. (Lond.) 221, 29–41 (1972)

    Google Scholar 

  • Donn, A., Maurice, D.M., Mills, N.L.: Studies on the living cornea in vitro. II The active transport of sodium across the epithelium. Arch. Ophthal. 62, 748–757 (1959)

    Google Scholar 

  • Ehlers, N.: Corneal adenosine triphosphatase: a histochemical investigation. Exp. Eye Res. 4, 48–53 (1965)

    Google Scholar 

  • Ernst, S.A.: Transport adenosine triphosphatase cytochemistry. I. Biochemical characterization of a cytochemical medium for the ultrastructural localization of ouabain-sensitive, potassium-dependent phosphatase activity in the avian salt gland. J. Histochem. Cytochem. 20, 13–22 (1972a)

    Google Scholar 

  • Ernst, S.A.: Transport adenosine triphosphatase. II. Cytochemical localization of ouabain-sensitive, potassium-dependent phosphatase activity in the secretory epithelium of the avian salt gland. J. Histochem. Cytochem. 20, 23–38 (1972b)

    Google Scholar 

  • Ernst, S.A.: Transport ATPase cytochemistry: ultrastructural localization of potassium-dependent and potassium-independent phosphatase activities in rat kidney cortex. J. Cell Biol. 66, 586–608 (1975)

    Google Scholar 

  • Firth, J.A.: Problems on the specificity in the use of a strontium capture technique for the cytochemical localization of ouabain-sensitive, potassium-dependent phosphatase in mammalian renal tubules. J. Histochem. Cytochem. 22, 1163–1168 (1974)

    Google Scholar 

  • Firth, J.A., Marland, B.Y.: The significance of inhibitor-resistant alkaline phosphatase in the cytochemical demonstration of transport adenosine triphosphatase. J. Histochem. Cytochem. 23, 571–574 (1975)

    Google Scholar 

  • Fischbarg, J., Lim, J.J.: Role of cations, anions and carbonic anhydrase in fluid transport across rabbit corneal endothelium. J. Physiol. (Lond.) 241, 647–675 (1974)

    Google Scholar 

  • Freeman, R.D.: Oxygen consumption by the component layers of the cornea. J. Physiol. (Lond.) 225, 15–32 (1972)

    Google Scholar 

  • Guth, L., Albers, R.W.: Histochemical demonstration of (Na+ -K+)-activated adenosine triphosphatase. J. Histochem. Cytochem. 22, 320–326 (1974)

    Google Scholar 

  • Hodson, S.: Evidence for a bicarbonate-dependent sodium pump in corneal endothelium. Exp. Eye Res. 11, 20–29 (1971)

    Google Scholar 

  • Jacobsen, N.O., Jörgensen, P.L.: A quantitative biochemical and histochemical study of the lead method for localization of adenosine triphosphate-hydrolyzing enzymes. J. Histochem. Cytochem. 17, 443–452 (1969)

    Google Scholar 

  • Kaye, G.I., Tice, L.W.: Studies on the cornea. V. Electron microscopic localization of adenosine triphosphatase activity in the rabbit cornea in relation to transport. Invest. Ophthal. 5, 22–31 (1966)

    Google Scholar 

  • Koenig, C.S., Vial, J.D.: A ciritical study of the histochemical lead method for localization of Mg-ATPase cell boundaries. Histochem. J. 5, 503–518 (1973)

    Google Scholar 

  • Kyte, J.: Immunoferritin determination of the distribution of (Na+ + K +) ATPase over the plasma membranes of renal convoluted tubules. I. Distal segment. J. Cell Biol. 68, 287–303 (1976a)

    Google Scholar 

  • Kyte, J.: Immunoferritin determination of the distribution of (Na+ + K+) ATPase over the plasma membranes of renal convoluted tubules. II. Proximal segment. J. Cell Biol. 68, 304–318 (1976b)

    Google Scholar 

  • Lambert, B., Donn, A.: Effect of ouabain on the active transport of sodium in the cornea. Arch. Ophthal. 72, 525–528 (1964)

    Google Scholar 

  • Langham, M.E., Kostelnik, M.: The effect of ouabain on the hydration and the adenosine triphosphatase activity of the cornea. J. Pharmacol. exp. Ther. 150, 398–405 (1965)

    Google Scholar 

  • Leuenberger, P.M., Novikoff, A.B.: Localization of transport adenosine triphosphatase in rat cornea. J. Cell Biol. 60, 721–731 (1974)

    Google Scholar 

  • Maeda, K., Sakaguchi, K.: Studies on sodium-potassium-activated adenosine triphosphatase in the cornea. Electron microscopic observations on the rat cornea. Jap. J. Ophthalmol. 9, 195–199 (1965)

    Google Scholar 

  • Marchesi, V.T., Palade, G.E.: The localization of Mg-Na-K-activated adenosine triphosphatase on red cell ghost membranes. J. Cell Biol. 35, 385–403 (1967)

    Google Scholar 

  • Matano, S., Ishii, S.: The fine structural localization of NaK-ATPase in the chorioid plexus in rat. J. Electron Micr. 22, 357–359 (1973)

    Google Scholar 

  • Maurice, D.M.:The location of the fluid pump in the cornea. J. Physiol. (Lond.) 221, 43–54 (1972)

    Google Scholar 

  • McClurkin, I.T.: A method for the cytochemical demonstration of sodium-activated adenosine triphosphatase. J. Histochem. Cytochem. 12, 654–658 (1964)

    Google Scholar 

  • Mishima, S., Kudo, T.: In vitro incubation of rabbit cornea. Invest. Ophthal. 6, 329–339 (1967)

    Google Scholar 

  • Palkama, A., Uusitalo, R.: Histochemical demonstration of ouabain-sensitive, sodium-potassiumactivated adenosine triphosphatase system. Scand. J. clin. Lab. Invest. (Suppl) 101, p. 48 (1968)

    Google Scholar 

  • Palkama, A., Uusitalo, R.: The histochemical demonstration of sodium-potassium-activated adenosine triphosphatase activity in rabbit ciliary body. Ann. Med. exp. Fenn. 48, 49–55 (1970)

    Google Scholar 

  • Palva, M., Palkama, A.: Histochemically demonstrable sodium-potassium-activated adenosine triphosphatase (Na-K-ATPase) activity in the rat lens. Exp. Eye Res. 19, 117–123 (1974)

    Google Scholar 

  • Palva, M., Palkama, A.: Electronmicroscopical, histochemical and biochemical findings on the Na-K-ATPase activity in the epithelium of the rat lens. Exp. Eye Res. 22, 229–236 (1976)

    Google Scholar 

  • Rogers, K.T.:Levels of (N+ + K+)-activated and Mg2+-activated ATPase activity in bovine and feline corneal endothelium and epithelium. Biochim. biophys. Acta (Amst.) 163, 50–56 (1968)

    Google Scholar 

  • Rosenthal, A.S., Moses, H.L., Ganote, C.E.: Interpretation of phosphatase cytochemical data. J. Histochem. Cytochem. 18, p. 915 (1970)

    Google Scholar 

  • Sabatini, M.T., Dipolo, R., Villegas, R.: Adenosine triphosphatase activity in the membranes of the squid nerve fibre. J. Cell Biol. 38, 176–183 (1968)

    Google Scholar 

  • Skou, J.C.: The influence of some cations on an adenosine triphosphatase from peripheral nerves. Biochim. biophys. Acta (Amst.) 23, 394–401 (1957)

    Google Scholar 

  • Skou, J.C.: Enzymatic basis for active transport of Na+ and K+ across cell membrane. Physiol. Rev. 45, 596–617 (1965)

    Google Scholar 

  • Stevenson, R.W., Wilson, W.S.: The effect of acetylcholine and eserine on the movement of Na+ across the corneal epithelium. Exp. Eye Res. 21, 235–244 (1975)

    Google Scholar 

  • Tervo, T.: Histochemical demonstration of cholinesterase activity in the cornea of the rat and the effect of various denervations on the corneal nerves. Histochemistry 47, 133–143 (1976)

    Google Scholar 

  • Tervo, T., Palkama, A.: Histochemical studies on adenosine triphosphatase activity in the rat cornea. Acta physiol. scand. 91, 21–22 (1974)

    Google Scholar 

  • Tervo, T., Palkama, A.: Electron microscopic localization of adenosine triphosphatase (NaKATPase) activity in the rat cornea. Exp. Eye Res. 21, 269–279 (1975)

    Google Scholar 

  • Thomas, R.C.: Electrogenic sodium pump in nerve and muscle cells. Physiol. Rev. 52, 563–592 (1972)

    Google Scholar 

  • Tormey, J. McD.: Significance of the histochemical demonstration of ATPase in epithelia noted for active transport. Nature (Lond.) 210, 820–822 (1966)

    Google Scholar 

  • Wachstein, M., Meisel, E.: Histochemistry of hepatic phosphatase at a physiologic pH. Amer. J. Path. 27, 13–23 (1957)

    Google Scholar 

  • Watson, M.L.: Staining of tissue sections for electron microscopy with heavy metals. J. biophys. biochem. Cytol. 4, 475–478 (1958)

    Google Scholar 

  • Whittam, R., Wheeler, K.P.: Transport across cell membranes. Ann. Rev. Physiol. 32, 21–60 (1970)

    Google Scholar 

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Authors and Affiliations

  1. Department of Anatomy, University of Helsinki, Helsinki, Finland

    Timo Tervo, Markku Palva & Arto Palkama

Authors
  1. Timo Tervo
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  2. Markku Palva
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  3. Arto Palkama
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Additional information

The work is part of an eye research project by Arto Palkama and supported by grants from the Sigrid Jusélius Foundation, Helsinki, Finland, to A.P. and from the Finnish Cultural Foundation, Helsinki, Finland, to T.T. and M.P.

The authors are grateful to Miss Irma Hiltunen for skilful technical assistance

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Tervo, T., Palva, M. & Palkama, A. Transport adenosine triphosphatase activity in the rat cornea. Cell Tissue Res. 176, 431–443 (1977). https://doi.org/10.1007/BF00231400

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