Abstract
Astroglial cell cultures were derived from newborn rat forebrain and cultured for 5 days in serum containing-, and for an additional 4 days in a serum-free, defined medium. At the end of this 9-day-long period, basic astroglial growth factor (AGF2) was administered to the culture medium (10 ng per ml). Cells were subsequently cultured in AGF2 containing serum-free, defined medium for further two weeks. At definite intervals of culturing, unidirectional influx of both Na+ and K+ (INa and IK, respectively) was determined by applying22Na and42K. The AGF2-treated cultures showed highly increased, amiloride-sensitive INa at the early exposure period (2–8 hours), similar to that we have reported about cultured astroglia exposed to AGF2 for minutes. They also exhibited significant furosemide-sensitive-, while relatively poor ouabain-sensitive component of INa. However, at later periods of exposure to AGF2, INa was significantly reduced, particularly due to the decrease of its amiloride-sensitive component, while its furosemide-sensitive component further increased with the time of AGF2 treatment. In contrast to INa, the IK in the cultures exposed to AGF2 increased significantly in the course of the long-term exposure period, particularly the ouabain-, and furosemide-sensitive-components, while its amiloride-sensitive component, similarly to that of INa, decreased. Our findings show that the initial activation of the Na+/H+ (or K+/H+) exchange, what characterized the cation transport changes by short-term exposure of astroglial cells to AGF2 in our previous study, comes relatively soon to a cessation but activation of the Na+, K+-pump and the furosemide-sensitive Na+ and K+ influxes further increases. Thus, they suggest the possibility that furosemide-sensitive cation movements play a role, besides the Na+, K+-pump, in the control of glial cell differentiation.
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References
Hertz, L., and Chaban, G. 1982. Indications for an active role of astrocytes in potassium homcostasis at the cellular level; Potassium uptake and metabolic effects of potassium. Pages 157–174,in Pfeiffer, S. E. (Ed.), Neuroscience Approached Through Cell Culture, CRC Press, Boca Raton, FL.
Hertz, L., Juurlink, B. H. J., and Szuchet, S. 1985. Cell cultures. Pages 603–661,in Lajtha, A. (Ed.), Handbook of Neurochemistry, Plenum Press, New York.
Hertz, L. 1986. Potassium transport in astrocytes and neurones in primary cultures. Annals N. Y. Acad. Sci. 481:318–333.
Waltz, W., and Hertz, L. 1983. Intracellular ion changes of astrocytes in response to extracellular potassium. J. Neurosci, Res. 10:411–423.
Waltz, W., Wuttke, W., and Hertz, L. 1984. Astrocytes in primary cultures; membrane potential characteristics reveal exclusive potassium conductance and potassium accumulator properties. Brain Research 292:367–374.
Waltz, W., and Hinks, E. 1986. A transmembrane sodium cycle in astrocytes. Brain Research 368:226–232.
Latzkovits, L., Torday, Cs., Labourdette, G., Pettmann, B., and Sensenbrenner, M. 1988. Sodium and potassium uptake in primary cultures of proliferating rat astroglial cells induced by shortterm exposure to an astroglial growth factor. Neurochem. Res. 13:837–848.
Moolenaar, W. H., Defize, L. H. K., Tilly, B. C., Bierman, A. J., and de Laat, S. W. 1986. Transmembrane signalling by growth factors. Annals N.Y. Acad. Sci. 488:491–502.
Leffert, H. M., and Koch, K. S. 1985. Growth regulation by sodium ion fluxes. Pages 367–413,in Boynton, A. L. and Leffert, H. L. (eds.). Control of Animal Cell Proliferation, Vol. 1, Academic Press, New York.
Rozengurt, E., and Mendoza, S.A. 1985. Synergistic signals in mitogenesis: role of ion fluxes, cyclic nucleotides and protein kinase C in swiss 3T3 cells. Pages 229–242,in Hopkins, C. R., and Hughes, R. C. (eds.), Growth Factors: Structure and Function, Supplement 3, J. Cell. Sci., Company of Biologists Limited, Cambridge.
Pouyssegur, J., Franchi, A., L'Allemain, G., and Paris, S. 1985. Cytoplasmic pH, a key determinant of growth factor induced DNA synthesis in quiescent fibroblasts. FEBS Lett. 190:115–119.
Moolenaar, W. H., Defize, L. H. K., and de, Laat, S. W. 1986. Ionic signalling by growth factor receptors. J. Exp. Biol. 124: 359–373.
Moolenaar, W. H., Boonstra, J., van der Saag, P. T., and de Laat, S. W. 1981. Sodium/proton exchange in mouse neuroblastoma cells. J. Biol. Chem. 256:12883–12887.
Moolenaar, W. H., Mummery, C. L., van der, Saag, P. T., and de Laat, S. W. 1981. Rapid ionic events and the initiation of growth in serum-stimulated neuroblastoma cells. Cell 23:789–798.
Moolenaar, W. H., Tsien, R. Y., van der Saag, P. T., and de Laat, S. W. 1983. Na+/H+ exchange and cytoplasmic pH in the action of growth factors in human fibroblasts. Nature 304:645–648.
Paris, S., and Pouyssegur, J. 1984. Growth factors activate the Na+/H+ antiporter in quiescent fibroblasts by increasing its affinity for intracellular H. J. Biol. Chem. 259:10989–10994.
Kimelberg, H. K., Biddlecome, S., and Bourke, R. S. 1989. SITS-inhibitable Cl− transport and Na+-dependent H+ production in primary astroglial cultures. Brain Res. 173:111–124.
Benos, D. J., Sapirstein, V. S. 1983. Characteristics of an amiloride-sensitive sodium entry pathway in cultured rodent glial and neuroblastoma cells. J. Cell. Physiol. 116:213–220.
Sapirstein, V. S., and Benos, D. J. 1984. Activation of amiloridesensitive sodium transport in C6 glioma cells. J. Neurochem. 43:1098–1105.
Jean, T., Frelin, C., Vigne P., and Lazdunski, M. 1986. The Na+/H+ exchange system in glial cell lines. Properties and activation by an hyperosmotic shock. Eur. J. Biochem. 160:211–219.
Boonstra, J., Moolenaar, W. H., Harrison, P. H., Moed, P., van der Saag, P. T., and de Laat, S. W. 1983. Ionic responses and growth stimulation induced by nerve growth factor and epidermal growth factor in rat pheochromocytoma (PC12) cells. J. Cell. Biol. 97:92–98.
Besterman, J. M., Tyrey, S. J., Cragoe, E. J. Jr., and Cuatrecasas, P. 1984. Inhibition of epidermal growth factor-induced mitogenesis by amiloride and an analog: evidence against a requirement for Na+/H+ exchange. Proc. Natl. Acad. Sci. USA 81:6762–6766.
Rozengurt, E., and Heppel, L. A. 1975. Serum rapidly stimulates ouabain-sensitive86Rb uptake in quiesscent 3T3 cells. Proc. Natl. Acad. Sci. USA 72:4492–4495.
Boonstra, J., Skaper, S. D., and Varon, S. 1982. Regulation of Na+, K+-pump activity by nerve growth factor in chick embryo dorsal root ganglion cells. J. Cell. Physiol. 113:28–34.
Skaper, S. D., and Varon, S. 1983. Control of the Na+, K+-pump by nerve growth factor is essential to neuronal survival. Brain Res. 271:263–271.
Smith, J. B., and Rozengurt, E. 1978. Serum stimulates the Na+, K+-pump in quiescent fibroblasts by increasing Na+ entry. Proc. Natl. Acad. Sci. USA 75:5560–5564.
Mendoza, S. A., Wigglesworth, N. M., Pirkko, P., and Rozengurt, E. 1980. Na+ entry and Na+, K+-pump activity in murine, hamster and human cells-effect of monensin, serum, platelet extract, and viral transformation. J. Cell. Comp. Physiol. 103:17–27.
Mitsumoto, Y., and Mohri, T. 1986. Leucine transport in relation to the activities of Na+-H+ antiporter and Na+, K+-pump stimulated by serum and tumor promoter. Biochim. Biophys. Acta 861:187–193.
O'Brien, T. G., and Krzeminski, K. 1983. Phorbol ester inhibits furosemide-sensitive potassium transport in BALB/c 3T3 preadipose cells. Proc. Natl. Acad. Sci. USA 80:4334–4338.
Sussman, I., and O'Brien, T. G. 1985. Characterization of a BALB/c 3T3 preadipose cell mutant with altered Na+K+Cl− cotransport activity. J. Cell. Physiol. 124:153–159.
O'Brien, T. G., Prettyman, R., George, K. S., and yHerschman, H. R. 1988. A phorbol ester-nonproliferative variant of swiss 3T3 cells is deficient in Na+K+Cl− cotransport activity. J. Cell. Physiol. 134:302–306.
Johnson, J. H., Dunn, D. P., and Rosenberg, R. N. 1982. Furosemide-sensitive K+ channel glial cells but not neuroblastoma cells in culture. Biochem. Res. Commun. 109:100–115.
Chassande, O., Frelin, C., Farahifar, D., Jean, T., and Lazdunski, M. 1988. The Na+/K+/Cl− cotransport in C6 glioma cells. Properties and role in volume regulation. Eur. J. Biochem. 171:425–433.
Sensenbrenner, M., Pettmann, B., Labourdette, G., and Weibel, M. 1985. Properties of a brain growth factor promoting proliferation and maturation of rat astroglial cells in culture. Pages 345–360,in Dumont, J. E., Hamprecht, B., and Nunez, J. (eds.), Hormones and Cell Regulation, INSERM European Symposium, Elsevier Science Publishers B. V., Biomedical Division.
Pettmann, B., Weibel, M., Daunc, G., Sensenbrenner, M., and Labourdette, G. 1982. Stimulation of proliferation and maturation of rat astroblasts in serum-free culture by an astroglial growth factor. J. Neurosci. Res. 8:463–476.
Weibel, M., Pettmann, B., Labourdette, G., Miehe, E., Bock, E., and Sensenbrenner, M. 1985. Morphological and biochemical maturation of rat astroglial cells grown in a chemically defined medium, influences of an astroglial growth factor. Int. J. Dev. Neurosci. 3:617–630.
Pettmann, B., Weibel, M., Sensenbrenner, M., and Labourdette, G. 1985. Purification of two astroglial growth factor from bovine brain. FEBS Lett. 189:102–108.
Pettmann, B., Labourdette, G., Devilliers, G., and Sensenbrenner, M. 1981. Effects of brain extracts from chick embryo on the development of astroblasts in culture. Dev. Neurosci. 4:37–45.
Sensenbrenner, M., Labourdette, G., Delaunoy, J. P., Pettmann, B., Devilliers, G., Moonen, G., and Bock, E. 1980. Morphological and biochemical differentiation of glial cells in primary culture. Pages 385–395,in Giacobini, E., Vernadakis, A., and Shahar, A. (eds.), Tissue Culture in Neurobiology, Raven Press, New York.
Pettmann, B., Labourdette, G., Weibel, M., and Sensenbrenner, M. 1985. Brain-derived astroglial growth factors Funkt. Biol. Med. 4:243–248.
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Special issue dedicated to Dr. Paola S. Timiras.
Cente de Neurochimie du CNRS.
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Latzkovits, L., Kátay, L., Torday, C. et al. Sodium and potassium uptake in primary cultures of rat astroglial cells induced by long-term exposure to the basic astroglial growth factor (AGF2). Neurochem Res 14, 1025–1030 (1989). https://doi.org/10.1007/BF00965938
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DOI: https://doi.org/10.1007/BF00965938