Abstract
The relationships between membrane fatty acid modification and neurite outgrowth and norepinephrine release were evaluated in PC12 cells. [3H]Norepinephrine release evoked by carbachol was unaffected by the modifications. Basal spontaneous release was elevated with increases in the degree of unsaturation using cells supplemented with n-3 fatty acids; a reverse correlation was observed for [3H]norepinephrine uptake. Supplementation of PC12 cells with either n-6 fatty acids or 18:1 also increased the basal release and decreased the uptake. Docosahexaenoic acid promoted and arachidonic acid suppressed neurite outgrowth induced by nerve growth factor. Choline acetyltransferase activity was slightly influenced by these fatty acids. Thus, modifications of PC12 cells with arachidonic acid and docosahexaenoic acid had a relatively small effect on the degree of differentiation but had pronounced but opposite effects on neurite elongation. Ethanolamine glycerophospholipid synthesis was elevated during differentiation induced by nerve growth factor and it was suppressed by added arachidonic acid but not by docosahexaenoic acid. Our results raise the possibility that the decreased phospholipid synthesis caused by arachidonate may lead to the suppression of neurite elongation.
Similar content being viewed by others
REFERENCES
Bazan, N. G. 1990. Supply of n-3 polyunsaturated fatty acids and their significance in the central nervous system. Pages 1–24, in Wurtman, R. J., and Wurtman, J. J. (eds.), Nutrition and the Brain. Vol. 8., Raven Press, New York.
Scott, B. L., and Bazan, N. G. 1989. Membrane docosahexaenoate is supplied to the developing brain and retina by the liver. Proc. Natl. Acad. Sci. USA 86:2903–2907.
Green, P., and Yavin, E. 1993. Elongation, desaturation, and esterification of essential fatty acids by fetal rat brain in vivo. J. Lipid Res. 34:2099–107.
Yamamoto, N., Saito, M., Moriuchi, A., Nomura, M., and Okuyama, H. 1987. Effect of the dietary α-linolenate/linoleate balance on lipid compositions and learning ability of rats. J. Lipid Res. 28:144–151.
Yamamoto, N., Hashimoto, A., Moriuchi, A., Takemoto, Y., Okuyama, H., Nomura, M., Kitajima, R., Togasi, T., and Tamal, Y. 1988. Effect of the dietary α-linolenate/linoleate balance on lipid compositions and learning ability of rats. II. Discrimination process, extinction process, and glycolipid compositions. J. Lipid Res. 29:1013–1021.
Yamamoto, N., Okaniwa, Y., Mori, S., Nomura, M., and Okuyama, H. 1991. Effect of the high α-linolenate diet on the learning ability of aged rats. J. Gelontol. 46:B17–22.
Watanabe, I., Kato, M., Aonuma, H., Hashimoto, A., Naito, Y., Moriuchi, A., and Okuyama, H. 1987. Effect of dietary α-linolenate/linoleate balance on the lipid composition and electroretinographic responses in rats. Adv. Biosci. 62:563–570.
Bourre, J. M., Francois, M., Youyou, A., Dumont, O., Piciotti, M., Pascal, G., and Durand, G. 1989. The effects of dietary α-linolenic acid on the composition of nerve membranes, enzymatic activity, amplitude of electrophysiological parameters, resistance to poisons and performance of learning tasks in rats. J. Nutr. 119:1880–1892.
Stubbs, C. D., and Smith, A. D. 1984. The modification of mammalian membrane polyunsaturated fatty acid composition in relation to membrane fluidity and function. Biochim. Biophys. Acta 779:89–137.
Tsutsumi, T., Yamauchi, E., Suzuki, E., Watanabe, S., Kobayashi, T., and Okuyama, H. 1995. Effect of a high α-linolenate and high linoleate diet on membrane-associated enzyme activities in rat brain—modulation of Na+, K−-ATPase activity at suboptimal concentrations of ATP. Biol. Pharm. Bull. 18:664–670.
Monck, J. R., and Fernandez, J. M. 1994. The exocytotic fusion pore and neurotransmitter release. Neuron 12:707–716.
Liscovitch, M., and Cantley, L. C. 1995. Signal transduction and membrane traffic: the PI-TP/phosphoinositide connection. Cell 81: 659–662.
Martin, R. E., and Bazan, N. G. 1992. Changing fatty acid content of growth cone lipids prior to synaptogenesis. J. Neurochem. 59: 318–325.
Yavin, E., and Yavin, Z. 1975. Polyunsaturated fatty acid metabolism in neuroblastoma cells in culture. J. Neurochem. 24:71–77.
Tixier-Vidal, A., Picart, R., Loudes, C., and Bauman, A. F. 1986. Effects of polyunsaturated fatty acids and hormones on synaptogenesis in serum-free medium cultures of mouse fetal hypothalamic cells. Neuroscience 17:115–132.
Greene, L. A., and Tischler, A. S. 1976. Establishment of a noradrenergic clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor. Proc. Natl. Acad. Sci. USA 73:2424–2428.
Greene, L. A., and Rein, G. 1977. Synthesis, storage and release of acetylcholine by a noradrenergic pheochromocytoma cell line. Nature 268:349–351.
Fonnum, F. 1975. A rapid radiochemical method for the determination of choline acetyltransferase. J. Neurochem. 24:407–409.
Bligh, E. G., and Dyer, W. J. 1959. A rapid method of total lipid extraction and purification. Can. J. Biochem. Physiol. 37:911–917.
Vance, J. E., Pan, D., Campenot, R. B., Bussiere, M., and Vance, D. E. 1994. Evidence that the major membrane lipids, except cholesterol, are made in axons of cultured rat sympathetic neurons. J. Neurochem. 62:329–337.
Chaves, E. P., Vance, D. E., Campenot, R. B., and Vance, J. E. 1995. Axonal synthesis of phosphatidylcholine is required for normal axonal growth in rat sympathetic neurons. J. Cell Biol. 128: 913–918.
Koda, Y., Wada, A., Yanagihara, N., Uezono, Y., and Izumi, F. 1989. cis-Unsaturated fatty acids stimulate catecholamine secretion, tyrosine hydroxylase and protein kinase C in adrenal medullary cells. Neuroscience 29:495–502.
Negishi, M., Ito, S., and Hayaishi, O. 1990. Arachidonic acid stimulates phosphoinositide metabolism and catecholamine release from bovine adrenal chromaffin cells. Biochem. Biophys. Res. Comm. 169:773–779.
Ehrengruber, M. U., and Zahler P. 1991. Inhibition of the nicotinic ion cannel by arachidonic acid and other unsaturated fatty acids in chromaffin cells from bovine adrenal medulla. Chimia 45:45–49.
Ehrengruber, M. U., Deranleau, D. A., Kempf, C., Zahler, P., and Lanzrein, M. 1993. Arachidonic acid and other unsaturated fatty acids alter membrane potential in PC12 and bovine adrenal chromaffin cells. J. Neurochem. 60:282–288.
Vaughan, P. F. T., Murphy, M. G., and Ball, S. G. 1993. Effect of inhibitors of eicosanoid metabolism on release of [3H]noradrenaline from the human neuroblastoma, SH-SY5Y. J. Neurochem. 60:1365–1371.
Friedrich, U., and Bonisch, H. 1986. The neuronal noradrenaline transport system of PC12 cells: kinetic analysis of the interaction between noradrenaline, Na+ and Cl− in transport. Naunyn-Schmeideburgs Arch. Pharmacol. 333:246–253.
Traynor, A., and Schubert, D. 1984. Phospholipases elevate cyclic AMP levels and promote neurite extension in a clonal nerve cell line. Develop. Brain Res. 14:197–204.
DeGeorge, J. J., Walenga, R., and Carbonetto, S. 1988. Nerve Growth Factor rapidly stimulates arachidonate metabolism in PC12 cells: potential involvement in nerve fiber growth. J. Neurosci. Res. 21:323–332.
Takahata, K. 1995. Effect of DHA on cultured neuronal cells: studies in PC12 cells. Jap. J. Pharmacol. 67(suppl. I):S13–3.
Murphy, E. J., and Horrocks, L. A. 1993. Effect of differentiation on the phospholipid and phospholipid fatty acid composition of N1E-115 neuroblastoma cells. Biochim. Biophys. Acta 1167:131–136.
Bazan, N. G., and Rodriguez, de Turco Eb. 1994. Review: pharmacological manipulation of docosahexaenoic-phospholipid biosynthesis in photoreceptor cells: implications in retinal degeneration. J. Ocul. Pharmacol. 10:591–604.
Gazzah, N., Gharib, A., Croset, M., Bobillier, P., Lagarde, M., and Sarda, N. 1995. Decrease of brain phospholipid synthesis in free-moving n-3 fatty acid deficient rats. J. Neurochem. 64:908–918.
Tsai, S. S., Sun, A. Y., Kim, H. D., and Sun, G. Y. 1993. Manganese exposure to PC-12 cells alters triacylglycerol metabolism and promotes neurite outgrowth. Life Sci. 19:1567–75.
Shinomura, T., Asaoka, Y., Oka, M., Yoshida, K., and Nishizuka, Y. 1991. Synergistic action of diacylglycerol and unsaturated fatty acid for protein kinase C activation: its possible implications. Proc. Natl. Acad. Sci. USA 88:5149–5153.
Newton, A. C., and Keranen, L. M. 1994. Phosphatidyl-L-serine is necessary for protein kinase C's high-affinity interaction with diacylglycerol-containing membranes. Biochemistry 33:6651–6658.
Kalman, J., Gecse, A., Farkas, T., Joo, F., Telegdy, G., and Lajtha, A. 1992. Dietary manipulation with high marine fish oil intake of fatty acid composition and arachidonic acid metabolism in rat cerebral microvessels. Neurochem. Res. 17:162–172.
Calderaro, V., Parrilol, C., Balestrieri, M. L., Giovane, A., Filippelli, A., and Rossi, F. 1994. Docosahexaenoic acid and signaling pathways in rabbit colon. Mol. Pharmacol. 45:737–746.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Ikemoto, A., Kobayashi, T., Watanabe, S. et al. Membrane Fatty Acid Modifications of PC12 Cells by Arachidonate or Docosahexaenoate Affect Neurite Outgrowth But Not Norepinephrine Release. Neurochem Res 22, 671–678 (1997). https://doi.org/10.1023/A:1027393724676
Issue Date:
DOI: https://doi.org/10.1023/A:1027393724676