Expression of the beta-galactoside alpha 1,2-fucosyltransferase gene suppresses axonal outgrowth of neuro2a neuroblastoma cells.
The axonal outgrowth of cells of Neuro2a, a mouse neuroblastoma cell line, was suppressed on expression of the beta-galactoside alpha 1,2-fucosyltransferase (alpha 1,2-FT) gene. We recently cloned two types of rabbit alpha 1,2-FT, RFT-I and RFT-II. RFT-I exhibits comparable kinetic properties and structural homology with human H gene alpha 1,2-FT, and RFT-II shows comparable kinetic parameters with human Se gene alpha 1,2-FT. Neuro2a cells expressing RFT-I (N2A-RFT-I) contained a large amount of fucosyl GM1 instead of GM1 and GD1a, major gangliosides in the parent Neuro2a cells, whereas Neuro2a cells expressing RFT-II (N2A-RFT-II) showed a subtle change in the ganglioside pattern. N2A-RFT-II and parent Neuro2a cells showed axonal outgrowth in serum-free medium on the exogenous addition of GM1, whereas N2A-RFT-I cells exhibited multiple neurite sprouts but not axonal outgrowth. This phenotype was fully recovered by N2A-RFT-I cells on the addition of D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol and alpha-L-fucosidase to the culture medium, which resulted in pronounced reduction of fucosyl GM1 expression. These results suggested that expression of H-type alpha1,2-FT, and subsequent incorporation of fucose into glycolipids and glycoproteins, especially the formation of fucosyl GM1, modifies the response of neuronal cells to stimuli that induce axonal extension.
Hitoshi
S
S
Institute of Physical and Chemical Research (RIKEN), Wako-shi, Saitama, Japan.
Koijima
N
N
Kusunoki
S
S
Inokuchi
J
J
Kanazawa
I
I
Tsuji
S
S
D000818:Animals; D001369:Axons / Q000201:enzymology*; D002454:Cell Differentiation / Q000502:physiology; D018076:DNA, Complementary / Q000235:genetics; D005647:Fucosyltransferases / Q000235:genetics*; D005732:Gangliosides / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology; D006023:Glycoproteins / Q000378:metabolism; D051379:Mice; D016501:Neurites / Q000502:physiology; D009447:Neuroblastoma / Q000201:enzymology*; D011817:Rabbits; D014162:Transfection; D014407:Tumor Cells, Cultured / Q000166:cytology / Q000201:enzymology; D000097631:Galactoside 2-alpha-L-fucosyltransferase
D018076:DNA, Complementary; D005647:Fucosyltransferases; D005732:Gangliosides; D006023:Glycoproteins; D000097631:Galactoside 2-alpha-L-fucosyltransferase