Immunohistochemical and biochemical analyses of GD3, GT1b, and GQ1b gangliosides during neural differentiation of P19 EC cells 1

FEBS Letters

Kotani, Masaharu¹  Takeda, Shigeki³  Osanai, Taka⁴  Kato, Hiroko⁵  Sanai, Yutaka⁴  Yuen, Chun-Ting² 

[1] Department of Clinical Genetics, The Tokyo Metropolitan Institute of Medical Science (RINSHOKEN), Tokyo Metropolitan Organization for Medical Research, 3-18-22 Honkomagome, Bunkyo-ku, Tokyo 113-8613, Japan;Laboratory for Molecular Structure, National Institute for Biological Standards and Control, Blanche Lane, South Mimms, Potters Bar, Hertfordshire EN6 3QG, UK;Niigata Neurosurgical Hospital, 3057 Yamada, Niigata, Niigata 950-1101, Japan;Department of Biochemical Cell Research, The Tokyo Metropolitan Institute of Medical Science (RINSHOKEN), Tokyo Metropolitan Organization for Medical Research, 3-18-22 Honkomagome, Bunkyo-ku, Tokyo 113-8613, Japan;Leica Microsystems, K.K., 1-11-2 Osaki, Shinagawa-ku, Tokyo 141-0032, Japan

关键词: Ganglioside;    Sialyltransferase;    Retinoic acid;    Neural cell differentiation;    Confocal laser scanning microscopy;    BSA;    bovine serum albumin;    CNS;    central nervous system;    EC;    embryonic carcinoma;    FITC;    fluorescein isothiocyanate;    Fr(s);    fraction(s);    Gal;    galactose;    Gal-T;    galactosyltransferase;    GAP-43;    growth-associated protein-43;    HPTLC;    high-performance thin-layer chromatography;    MAb;    monoclonal antibody;    NCAMs;    neural cell adhesion molecules;    NeuAc;    N-acetylneuraminic acid;    RA;    retinoic acid;    R/A;    RA-induced aggregates;    R/N;    RA-induced neurons;    SAT(s);    sialyltransferase(s);    TBS-T;    Tris-buffered saline-Tween 20;    U;    undifferentiated cells;   

DOI  :  10.1016/S0014-5793(03)00083-8

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

In an earlier study, we showed that expressions of GD3, GT1b, and GQ1b gangliosides in P19 embryonic carcinoma (EC) cells were enhanced during their neural differentiation induced by retinoic acid. We now further demonstrated that this increase of the b-series gangliosides is due to an increase in their corresponding synthases (sialyltransferase-II, -IV, and -V) in the Golgi. Of the three gangliosides studied, GQ1b appeared to be the best candidate for monitoring such differentiation process. We also used fluorescence-labeled monoclonal antibodies and confocal fluorescence microscopy to obtain direct visual information about the relationship of gangliosides and neural specific proteins in neuron development. Again, GQ1b is the most interesting as it localizes with synaptophysin and neural cell adhesion molecules (NCAMs) on synaptic boutons or dendritic spines in RA-induced neurons (R/N). This suggests that GQ1b could be used as a marker for synapse formation during construction of the neural network.

【 授权许可】

Unknown