Expression of glycoconjugates bearing the Lewis X epitope during neural differentiation of P19 EC cells

FEBS Letters

Shimoda, Yasushi¹  Chai, Wengang³  Osanai, Taka¹  Sanai, Yutaka¹  Tajima, Youichi¹  Yuen, Chun-Ting² 

[1] Department of Biochemical Cell Research, Tokyo Metropolitan Institute of Medical Science (RINSHOKEN), 18-22, Honkomagome 3-chome, Bunkyo-ku, Tokyo 113-8613, Japan;Laboratory of Molecular Structure, National Institute for Biological Standards and Control, Blanche Lane, South Mimms, Potters Bar, Herts EN6 3QG, UK;MRC Glycosciences Laboratory, Imperial College School of Medicine, Northwick Park Hospital, Watford Road, Harrow, Middlesex HA1 3UJ, UK

关键词: Retinoic acid;    Lex;    Fucosyltransferase;    Fuc-TIX gene;    Neural cell differentiation;    Embryonic carcinoma cell;    BSA;    bovine serum albumin;    DHPE;    L-1;    2-dihexadecyl-sn-glycero-phosphoethanolamine;    EC;    embryonic carcinoma;    Fuc;    fucose;    Fuc-T(s);    fucosyltransferase(s);    mFuc-T(s);    mouse fucosyltransferase(s);    Hex;    hexose;    dHex;    deoxyhexose;    HexNAc;    N-acetylhexosamine;    HPLC;    high-performance liquid chromatography;    HPRT;    hypoxanthine phosphoribosyltransferase;    Lex;    Lewis X (3-fucosyl N-acetyllactosamine sequence);    LSIMS;    liquid secondary ion mass spectrometry;    MAb;    monoclonal antibody;    PA;    pyridylaminated;    PBS;    phosphate-buffered saline;    RA;    retinoic acid (all-trans form);    RT;    reverse transcriptase;    RT-PCR;    reverse transcription-polymerase chain reaction;    TLC;    thin-layer chromatography;    UV;    ultraviolet light;    Cer;    ceramide;    CMH;    Galβ1-1Cer;    CDH;    Galβ1-4Glcβ1-1Cer;    CTH;    Galα1-4Galβ1-4Glcβ1-1Cer;    Forssman;    GalNAcα1-3GalNAcβ1-3Galα1-4Galβ1-4Glcβ1-1Cer;    GA1;    asialo-GM1;    Galβ1-3GalNAcβ1-4Galβ1-4Glcβ1-1Cer;    globoside;    GalNAcβ1-3Galα1-4Galβ1-4Glcβ1-1Cer;    paragloboside;    Galβ1-4GlcNAcβ1-3Galβ1-4Glcβ1-1Cer;    Oligosaccharides used were: LNFP-III;    lacto-N-fucopentaose-III;    Galβ1-4(Fucα1-3)GlcNAcβ1-3Galβ1-4Glc;    LNnT;    lacto-N-neotetraose;    Galβ1-4GlcNAcβ1-3Galβ1-4Glc2;   

DOI  :  10.1016/S0014-5793(00)02407-8

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The Lewis X (Le^x) bearing glycolipids were noticeably increased in amounts during the course of neural differentiation of P19 EC cells induced by retinoic acid (RA, all-trans form). Applying neoglycolipid technology and in situ TLC-LSIMS, the oligosaccharide chains of these scarce Le^x bearing glycolipids were partially characterized after released by endoglycoceramidase and subsequent conversion into neoglycolipids. In order to understand the enzymatic basis for the expression of Le^x bearing glycolipids, we measured glycolipid, glycoprotein and oligosaccharide fucosyltransferase (Fuc-T) activities using appropriate substrates in P19 EC cells with or without RA treatment. All three Fuc-Ts were increased after RA treatment and the highest activity was in the differentiated neural cells. We then investigated the two possible Fuc-T genes that might be responsible for these changes using RT-PCR analysis. Mouse Fuc-TIX (mFuc-TIX) transcript was detected in all cell types but it was only strongly expressed in RA-induced aggregates and neural cells. In the case of mouse Fuc-TIV (mFuc-TIV) gene, its transcript was only detectable in RA-induced aggregates and not found in either undifferentiated or RA-induced neural cells. These results strongly support that RA induces only a transient expression of the mFuc-TIV gene in cell aggregates but a more persistent expression of the mFuc-TIX gene at the transcription level throughout neural cell differentiation. The mFuc-TIX gene is probably the main cause for the increased expression of Le^x glycoconjugates during neural differentiation of P19 EC cells.

【 授权许可】

Unknown