Abstract
This study focuses on the oxygen-dependence of active and passive K+ fluxes across membranes of cerebellar granule cells of neonatal rats. Maximal Na+,K+-ATPase activity along with minimal passive K+ influx was observed within oxygen concentration range characteristic for neonatal rat cerebellum. Prolonged exposure to hypoxia as well as hyperoxia resulted in suppression of the Na+,K+-ATPase and activation of the passive K+ flux. Toxic effects of hypoxia could be partially prevented by inhibition of NO production with L-NAME. This was accomplished by suppression of Na+,K+-ATPase with subsequent reduction in ATP consumption concurrently with the reduction in passive K+ flux. Activation of the Na+,K+-ATPase by NO at physiological pO2 could be abolished by inhibition of NO synthase by L-NAME or soluble guanylyl cyclase with ODQ. However, treatment of cells with activator of PKG Rp-8-CTP did not mimic normoxic activation of the active K+ influx. Oxygen-induced responses under normoxic conditions were differentially mediated by α1 isoform of the Na+,K+-ATPase catalytic subunit, whereas α2/3 isoform was predominantly active under conditions of severe hypoxia. We conclude that both hypoxia and hyperoxia trigger a gradual dissipation of transmembrane K+ gradient and loss of excitability of cerebellar neurons. The latter may be partially reversed by suppression of NO production under hypoxic conditions
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Hochachka, P.W. and Lutz, P.L., Mechanism, Origin, and Evolution of Anoxia Tolerance in Animals, Comp. Biochem. Physiol. B, Biochem. Mol. Biol., 2001, vol. 130, pp. 435–459.
Nilsson, G.E. and Lutz, P.L., Anoxia Tolerant Brains, J. Cereb. Blood Flow Metab., 2004, vol. 24, pp. 475–486.
Erecinska, M. and Silver, I.A., Tissue Oxygen Tension and Brain Sensitivity to Hypoxia, Respir Physiol., 2001, vol. 128, pp. 263–276.
Bogdanova, A., Petrushanko, I., Boldyrev, A., and Gassmann, M., Oxygen- and Redox-Induced Regulation of the Na/K ATPase, Curr Enzyme Inhibition, 2006, vol. 2, pp. 37–59.
Buck, L.T. and Hochachka, P.W., Anoxic Suppression of Na+-K+-ATPase and Constant Membrane Potential in Hepatocytes: Support for Channel Arrest, Am. J. Physiol., 1993, vol. 265, pp. R1020–R1025.
Dada, L.A., Chandel, N.S., Ridge, K.M., Pedemonte, C., Bertorello, A.M., and Sznajder, J.I., Hypoxia-Induced Endocytosis of Na,K-ATPase in Alveolar Epithelial Cells Is Mediated by Mitochondrial Reactive Oxygen Species and PKC-zeta, J. Clin. Invest., 2003, vol. 111, pp. 1057–1064.
Bogdanova, A., Grenacher, B., Nikinmaa, M., and Gassmann, M., Hypoxic Responses of Na/K-ATPase in Trout Hepatocyte Primary Cultures, J. Exp. Biol., 2005, vol. 208, pp. 1793–1803.
Khan, A.I., Drew, C., Ball, S.E., Ball, V., Ellory, J.C., and Gibson, J.S., Oxygen Dependence of K+-Cl- Cotransport in Human Red Cell Ghosts and Sickle Cells, Bioelectrochemistry, 2004, vol. 62, pp. 141–146.
Petrushanko, I.Y., Bogdanov, N.B., Lapina, N., Boldyrev, A., Gassmann, M., and Bogdanova, A.Y.u., Oxygen-Induced Regulation of Na/K-ATPase in Cerebellar Granule Cells, J. Gen, Physiol., 2007, vol. 130(4), pp. 389–398.
Comellas, A.P., Dada, L.A., Lecuona, E., Pesce, L.M., Chandel, N.S., Quesada, N., Budinger, G.R., Strous, G.J., Ciechanover, A., and Sznajder, J.I., Hypoxia-Mediated Degradation of Na,K-ATPase via Mitochondrial Reactive Oxygen Species and the Ubiquitin-Conjugating System, Circ. Res., 2006, vol. 98, pp. 1314–1322.
Petrushanko, I., Bogdanov, N., Bulygina, E., Grenacher, B., Leinsoo, T., Boldyrev, A., Gassmann, M., and Bogdanova, A., Na-K-ATPase in Rat Cerebellar Granule Cells Is Redox Sensitive, Am. J. Physiol. Regul. Integr Comp. Physiol., 2006, vol. 290, pp. R916–R925.
Rathbun, W.B. and Betlach, M.V., Estimation of Enzymically Produced Orthophosphate in the Presence of Cysteine and Adenosine Triphosphate, Anal. Biochem., 1969, vol. 28, pp. 436–445.
Cortes, A., Cascante, M., Cardenas, M.L., and Cornish-Bowden, A., Relationships between Inhibition Constants, Inhibitor Concentrations for 50% Inhibition and Types of Inhibition: New Ways of Analysing Data, Biochem. J., 2001, vol. 357, pp. 263–268.
Palasis, M., Kuntzweiler, T.A., Arguello, J.M., and Lingrel, J.B., Ouabain Interactions with the H5-H6 Hairpin of the Na,K-ATPase Reveal a Possible Inhibition Mech anism via the Cation Binding Domain, J. Biol. Chem., 1996, vol. 271, pp. 14176–14182.
Roy, B. and Garthwaite, J., Nitric Oxide Activation of Guanylyl Cyclase in Cells Revisited, Proc. Natl. Acad. Sci. USA, 2006, vol. 103, pp. 12185–12190.
Ogunshola, O.O., Stewart, W.B., Mihalcik, V., Solli, T., Madri, J.A., and Ment, L.R., Neuronal VEGF Expression Correlates with Angiogenesis in Postnatal Develop ing Rat Brain, Brain Res. Dev. Brain Res., 2000, vol. 119, pp. 139–153.
Lingrel, J., Moseley, A., Dostanic, I., Cougnon, M., He, S., James, P., Woo, A., O'Connor, K., and Neumann, J., Functional Roles of the Alpha Isoforms of the Na,K-ATPase, Ann. N. Y. Acad. Sci., 2003, vol. 986, pp. 354–359.
Scorziello, A., Pellegrini, C., Secondo, A., Sirabella, R., Formisano, L., Sibaud, L., Amoroso, S., Canzoniero, L.M., Annunziato, L., and Di Renzo, G.F., Neuronal NOS Activation During Oxygen and Glucose Deprivation Triggers Cerebellar Granule Cell Death in the Later Reoxygenation Phase, J. Neurosci. Res., 2004, vol. 76, pp. 812–821.
Jesko, H., Chalimoniuk, M., and Strosznajder, J.B., Activation of Constitutive Nitric Oxide Synthase(s) and Absence of Inducible Isoform in Aged Rat Brain, Neurochem. Int., 2003, vol. 42, pp. 315–322.
Guo, Y., Ward, M.E., Beasjours, S., Mori, M., and Hussain, S.N., Regulation of Cerebellar Nitric Oxide Production in Response to Prolonged In vivo Hypoxia, J. Neurosci. Res., 1997, vol. 49, pp. 89–97.
Matsumoto, T., Pollock, J.S., Nakane, M., and Forstermann, U., Developmental Changes of Cytosolic and Particulate Nitric Oxide Synthase in Rat Brain, Brain Res. Dev. Brain. Res., 1993, vol. 73, pp. 199–203.
Barjavel, M.J. and Bhargava, H.N., Nitric Oxide Synthase Activity in Brain Regions and Spinal Cord of Mice and Rats: Kinetic Analysis, Pharmacology, 1995, vol. 50, pp. 168–174.
Chalimoniuk, M. and Strosznajder, J.B., Aging Modulates Nitric Oxide Synthesis and cGMP Levels in Hippocampus and Cerebellum. Effects of Amyloid Beta Peptide, Mol. Chem. Neuropathol., 1998, vol. 35, pp. 77–95.
Wang, W., Nakayama, T., Inoue, N., and Kato, T., Quantitative Analysis of Nitric Oxide Synthase Expressed in Developing and Differentiating Rat Cerebellum, Brain Res. Dev. Brain Res., 1998, vol. 111, pp. 65–75.
Halcak, L., Pechanova, O., Zigova, Z., Klemova, L., Novacky, M., and Bernatova, I., Inhibition of NO Synthase Activity in Nervous Tissue Leads to Decreased Motor Activity in the Rat, Physiol. Res., 2000, vol. 49, pp. 143–149.
Bogdanova, A., Bogdanov, N., Boldyrev, A., Gassmann, M., and Pertushanko, I., Oxygen-Induced Regulation of the Na/K-ATPase in Dissociated Cerebellar Granule Cells, Free Rad. Res., 2006, vol. 40, p. S75.
Ellis, D.Z. and Sweadner, K.J., NO Regulation of Na,KATPase: Nitric Oxide Regulation of the Na,K-ATPase in Physiological and Pathological States, Ann. N. Y. Acad. Sci., 2003, vol. 986, pp. 534–535.
Boldyrev, A.A., Bulygina, E.R., Kramarenko, G.G. and Vanin, A.F. Effect of Nitroso Compounds on Na/K-ATPase, Biochim Biophys Acta, 1997, vol. 1321, pp. 243–251.
Kocak-Toker, N., Giris, M., Tulubas, F., Uysal, M., and Aykac-Toker, G., Peroxynitrite Induced Decrease in Na+,K+-ATPase Activity Is Restored by Taurine, World J. Gastroenterol., 2005, vol. 11, pp. 3554–3557.
Beltowski, J., Marciniak, A., Jamroz-Wisniewska, A., and Borkowska, E., Nitric Oxide-Superoxide Cooperation in the Regulation of Renal Na+,K+-ATPase, Acta Biochim. Pol., 2004, vol. 51, pp. 933–942.
Gupta, S., Chough, E., Daley, J., Oates, P., Tornheim, K., Ruderman, N.B., and Keaney, J.F. Jr., Hyperglycemia Increases Endothelial Superoxide that Impairs Smooth Muscle Cell Na+,K+-ATPase Activity, Am. J. Physiol. Cell Physiol., 2002, vol. 282, pp. C560–C566.
Gupta, S., Mcarthur, C., Grady, C., and Ruderman, N.B., Stimulation of Vascular Na+-K+-ATPase Activity by Nitric Oxide: A cGMP-Independent Effect, Am. J. Physiol., 1994, vol. 266, pp. H2146–H2151.
Gupta, S., Phipps, K., and Ruderman, N.B., Differential Stimulation of Na' Pump Activity by Insulin and Nitric Oxide in Rabbit Aorta, Am. J. Physiol., 1996, vol. 270, pp. H1287–H1293.
Zhou, L., Burnett, A.L., Huang, P.L., Becker, L.C., Kuppusamy, P., Kass, D.A., Kevin Donahue, J., Proud, D., Sham, J.S., Dawson, T.M., and Xu, K.Y., Lack of Nitric Oxide Synthase Depresses Ion Transporting Enzyme Function in Cardiac Muscle, Biochem. Biophys. Res. Commun., 2002, vol. 294, pp. 1030–1035.
Chalimoniuk, M., Glowacka, J., Zabielna, A., Eckert, A., and Strosznajder, J.B., Nitric Oxide Alters Arachidonic Acid Turnover in Brain Cortex Synaptoneurosomes, Neurochem. Int., 2006, vol. 48, pp. 1–8.
Segall, L., Javaid, Z.Z., Carl, S.L., Lane, L.K., and Blostein, R., Structural Basis for Alphal Versus Alpha2 Isoform-Distinct Behavior of the Na,K-ATPase, J. Biol. Chem., 2003, vol. 278, pp. 9027–9034.
Dobretsov, M. and Stimers, J.R., Neuronal Function and Alpha3 Isoform of the Na/K-ATPase, Front. Biosci., 2005, vol. 10, pp. 2373–2396.
Doll, C.J., Hochachka, PW., and Reiner, PB., Channel Arrest: Implications from Membrane Resistance in Turtle Neurons, Am. J. Physiol., 1991, vol. 261, pp. R1321–R1324.
Author information
Authors and Affiliations
Corresponding author
Additional information
Original Russian Text © N.B. Bogdanov, I.Yu. Petrushanko, A.A. Boldyrev, M. Gassmann, A.Yu. Bogdanova, 2008, published in Biologicheskie Membrany, 2008, Vol. 25, No. 1, pp. 25–33.
Rights and permissions
About this article
Cite this article
Bogdanov, N.B., Petrushanko, I.Y., Boldyrev, A.A. et al. Oxygen-sensitivity of potassium fluxes across plasma membrane of cerebellar granule cells. Biochem. Moscow Suppl. Ser. A 2, 26–32 (2008). https://doi.org/10.1134/S1990747808010054
Received:
Published:
Issue Date:
DOI: https://doi.org/10.1134/S1990747808010054