Abstract
The kinetics of manganese(II) ion uptake and efflux have been investigated using tracer54Mn(II) with glial cells cultured from chick cerebral cortex in chemically defined medium. The initial velocity of Mn(II) uptake versus [Mn(II)] exhibit saturation, with an apparent S0.5≈18(±3) μM. Both the rate and extent of Mn(II) uptake are inhibited by Ca(II), either added externally or preloaded into the glial cells. Preloading of glia with Mn(II) also inhibits the rate of external54Mn(II) uptake. Zn(II) inhibits but Cu(II) activates Mn(II) uptake. Efflux of Mn(II) from preloaded cells occurs as a biphasic process, with rapid release of 30–40% of total cell Mn(II), then much slower release of the remainder. Permeabilization of cells with dextran sulfate also rapidly released ca. 30% of total cell Mn(II). High external Mn(II) enhanced both the rate and extent of Mn(II) efflux. CCCP, an uncoupler of oxidative phosphorylation, inhibited both Mn(II) uptake and efflux significantly, but addition of cyanide, ouabain, insulin, hydrocortisone, K+, or Nd(III) had no effect on either process. Taken together, these data suggest a model in which Mn(II) is brought across the plasma membrane by facilitated diffusion, binds to cytosolic protein sites, and is partitioned into the mitochondria by an active transport mechanism. The fact that the Mn(II) flux rates observed with cultured glia are much faster than those reported for overall uptake and efflux of brain Mn(II)in vivo suggests that the blood-brain barrier may play a significant role in determining these latter rates in whole animals.
Similar content being viewed by others
References
Orent, E. R., and McCollum, E. V. 1931. Effects of deprivation of manganese in the rat. J. Biol. Chem. 92:651–678.
Metal Ions in Neurology and Psychiatry Pages 3–34, 49–68, 121–128,in Gabay, S., harris, J., and Ho, B. T., (eds.) Liss, New York, 1985.
Underwood, E. J. 1977. Trace Elements in Human and Animal Nutrition. 4th Edn. Pages 170–195, Academic Press, New York.
Manganese in Metabolism and Enzyme Function in Schramm, V. L., and Wedler, F. C., (eds.) Academic Press, New York, 1986.
Gianutsos, G., Seltzer, M. D., Saymeh, R., Wu, M. W., Michel, R. G. 1985. Brain manganese accumulation following systemic administration of different forms. Arch Toxicol. 57:272–275.
Bonilla, E., Levine, S., and DeSalazar, E. 1978. Intoxicacion cronica con manganeso. Acta Ciet. Venezolana. 29:332–337.
Mena, I., Marin, O., Fuenzalida, S., and Cotzias, G. C. 1967. Chronic manganese poisoning—clinical picture and manganese turnover. Neurology. 17:128–136.
Parenti, M., Flauto, C., Parati, E., Vescovi, A., and Groppetti, A. 1986. Manganese neuro-toxicity: effects of L-DOPA and pargyline treatments. Brain Research. 367:8–13.
Kristensson, K., Eriksson, H., Lundh, B., Plantin, L.-O., Wachtmeister, L., elAzazi, M., Morath, C., and Heilbrom, E. 1986. Effects of MnCl2 on the rat developing nervous system. Acta Pharmacol. et Toxicol. 59:345–348.
Hurley, L. S., Woolley, D. E., Rosenthal, F., and Timiras, P. S. 1963. Influence of manganese on susceptibility of rats to convulsions. Am. J. Physiol. 204:493–496.
Carl, G. F., Keen, C. L., Gallagher, B. B., Clegg, M. S., Littleton, W. H., Flannery, D. B., and Hurley, L. S. 1986. Association of low blood Mn concentrations with epilepsy. Neurology. 36:1584–1587.
Schramm, V. L. 1982. Metabolic regulation: could Mn2+ be involved? Trends Biochem. Sci. 7:369–371.
Williams, R. J. P. 1982. Free manganese (II) and iron (II) cations can act as intracellular cell controls. FEBS Lett. 140:3–10.
McCord, J. M., Boyle, J. A., Day, E. D., Rizollo, L. J., and Salin, M. L. 1977. Michaelson, A. M., McCord, J. M., & Fridovich, I., (eds.)in Superoxide and Superoxide Dismutases Page 129, Academic Press, New York.
Doherty, J. D., Salem, N., Lauter, C. J., and Trams, E. G. 1983. Mn2+-Stimulated ATPase in Rat Brain. Neurochem. Res. 8:493–499.
Wedler, F. C. and Toms, R. 1986. Interactions of Mn(II) with Mammalian Glutamine Synthetase. Pages 221–238,in Schramm, V. L., and Wedler, F. C., (eds.) Manganese in Metabolism and Enzyme Function Academic Press, New York.
Metabolic Compartmentation in the Brain (Balazs, R. and Cremer, J. E., eds.). Pages 167–184. MacMillan Press, London, 1973.
Norenberg, M., and Martinez-Hernandez, A. 1979. Fine structural localization of glutamine synthetase in astrocytes of rat brain. Brain Research. 161:303–310.
Walker, J. E. 1983. Glutamate, GABA, and CNS Disease: A Review. Neurochem. Res. 8:521–548.
Tholey, G., Ledig, M., Mandel, P., Sargentini, L., Frivold, A. H., Leroy, M., Grippo, A. A., and Wedler, F. C. 1987. Concentrations of physiologically important metal ions in glial cells cultured from chick cerebral cortex. Neurochem. Res. 12:45–50.
Schramm, V. L., and Brandt, M. 1986. The manganese(II) economy of rat hepatocytes. Fed. Proc. 45:2817–2820.
Stein, W. D. Transport and Diffusion across Cell Membranes, Academic Press, New York, 1986.
Tholey, G., Ledig, M., Kopp, P., Sargentini-Maier, L., Leroy, M., Grippo, A. A., and Wedler, F. C. 1988. Levels and subcellular distribution of physiologically important metal ions in neuronal cells cultured from chick embryo cerebral cortex. Neurochem. Res. 13:1163–1167.
Schramm, V. L. 1985. Evaluation of Mn(II) in metabolic regulation: Analysis of proposed sites for regulation. Pages 1109–1132,in (Schramm, V. L., & Wedler, F. C., eds.) “Manganese in Metabolism and Enzyme Function” Academic Press, New York.
Sensenbrenner, M. 1977. Dissociated brain cells in primary culture, Pages 191–213,in Federoff, S. and Hertz, L., (eds.) Cell, Tissue and Organ Culture in Neurobiology, Academic Press, New York.
Bottenstein, J. E., and Sato, G. H. 1979. Growth of a neuroblastoma cell line in serum-free supplemented medium. Proc. Nat. Acad. Sci., USA. 76:514–517.
Kimelberg, H. K., and Frangakis, M. V. 1985. Furosemide- and bumetimide-sensitive ion transport and volume control in primary astrocyte cultures from rat brain. Brain Res. 361:125–134.
Lowry, O. H., Rosenbrough, N. J., Farr, A. L., and Randall, R. J. 1951. Protein measurements with the Folin phenol reagent. J. Biol. Chem. 193:265–275.
Allshire, A., and Saris, N.-E. L. 1986. Interaction of manganese with cellular calcium homeostasis. Pages 51–64,in Schramm, V. L., & Wedler, F. C., (eds.) Manganese in Metabolism and Enzyme Function Academic Press, New York.
Shamoo, A. E. 1986. Mn(II) and Ca(II) transport in mitochondria. Pages 65–80,in Schramm, V. L., and Wedler, F. C., (eds.) Manganese in Metabolism and Enzyme Function Academic Press, New York.
Brandt, M., and Schramm, V. L. 1986. Mammalian manganese metabolism and manganese uptake and distribution in rat hepatocytes. Pages 3–16,in Schramm, V. L., and Wedler, F. C., (eds.) Manganese in Metabolism and Enzyme Function Academic Press, New York.
Smyers-Verbeke, J., May, C., Drochmans, P. and Massart, D. L. 1977. The Determination of copper, zinc and manganese in subcellular rat liver fractions. Anal. Biochem. 83:746–753.
Konji, V., Montag, A., Sandri, G., Nordenbrand, K., and Ernster, L. 1985. Transport of calcium and manganese by mitochondria from rat liver, heart and brain. Biochimie. 67:1241–1250.
Sakurai, H., Nishida, M., Yoshimura, T., Takada, J., and Koyama, M. 1985. Partition of divalent and total Mn in organs and subcellular organelles of MnCl2-treated rats studied by esr and neutron activation analysis. Biochem. Biophys. Acta. 841:208–214.
Suzuki, H., and Wada, O. 1981. Role of liver lysosomes in uptake and biliary excretion of manganese in mice. Environmental Research 26:521–528.
Goldstein, G. W., and Betz, A. L. 1986. The blood-brain barrier. Scientific Amer. 255:74–83.
Betz, A. L. 1986. Transport of ions across the blood-brain barrier. Fed. Proc. 45:2050–2054.
Betz, A. L., Goldstein, G. W., and Katzman, R. 1989. Blood-Brain-Cerebrospinal Fluid Barriers, Pages 591–608,in Siegel, G. J., et al. (eds.) Basic Neurochemistry, 4th Ed. Raven Press, New York.
Author information
Authors and Affiliations
Additional information
Supported in part by NIH grant GM-33358 and a Biomedical Research Support Grant from the NIH administered by Penn State.
Rights and permissions
About this article
Cite this article
Wedler, F.C., Ley, B.W. & Grippo, A.A. Manganese(II) dynamics and distribution in glial cells cultured from chick cerebral cortex. Neurochem Res 14, 1129–1135 (1989). https://doi.org/10.1007/BF00965619
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00965619