Ganglioside composition of growth cone-deficient nerve cell cultures.

Abstract

ABSTRACT. The ganglioside concentration and composition in growth cone-deficient nerve cells, induced by inclusion of cytochalasin B (CB) are compared with those of 2-day-old control cells from primary cultures of embryonic rat cerebral cortex. Ganglioside GM1 and GD 1 a are the major gangliosides in the growth cone. Ganglioside GM1 may be one of the membrane components of growth cones that function in neural recognition during development.Nerve growth cones are essential for establishing the appropriate neural connections during development because this organelle is the part of the neuron that recognizes its target cell. Which molecules mediate cell-cell recognition are not known, but gangliosides are major membrane constituents that may function in cell growth regulation and cellular interaction (9, 10,17).Sufficient numbers of growth cones must be obtained in order to investigate their biochemical constituents. In our identification of these constituents, we used growth cone-deficient nerve cell cultures induced by the inclusion of cytochalasin B (CB) in the culture medium. The biochemistry of these deficient cultures have been compared to normal control cultures (13), in order to determine which ganglioside components are localized specifically in growth cones. MATERIALS AND METHODSNeuron-enriched primary cultures of embryonic rat cerebral cortex were prepared as described previously (2, 3). Cone-deficient nerve cell cultures from embryonic rat cerebral cortex were made by adding cytochalasin B (5 µg/ml) to the culture medium according to the method of Marsh and Letourneau (13), after which the cells were cultured for 2 days in 3 % CO2 and 97 % air at 36.5°C.Biotinylated choleragen, used to identify ganglioside GM1, was prepared as described previously (1, 4). Briefly, 2 mg of the choleragen B subunit from a 1-ml solution of 50 mM Tris-HC1, 0.2 M NaCl and 1 mM Na2 EDTA at pH 8.0 was biotinylated with 30 pi of Correspondence to: Hiroaki Asou, Department of Physiology, Toho University, School of Medicine, 5-21-16, Ohmori-nishi,

Expression of gangliosides in neuronal development of P19 embryonal carcinoma stem cells

“…Therefore, the gangliosides stained by cholera toxin B subunit in neurites and growth cones of Day 6 are most likely of the ganglio-N-tetraose series consisting of GD1a, GT1a, GT1b and GQ1b, and the gangliosides detected in the fully grown neurites are most likely GD1b, GT1b and GQ1b. The results are consistent with several other studies showing the presence of gangliosides in isolated growth cone-enriched fractions (Asou and Hirano, 1987;Sbaschnig-Agler et al, 1988;Rosner et al, 1993;Igarashi et al, 1994;Meiri and Beverly, 1994) and further reveal that these gangliosides consist primarily of the ganglio-Ntetraose series.…”