Gangliosides are known to be important in many biological processes. However, details concerning the exact function of these glycosphingolipids in cell physiology are poorly understood. In this study, the role of gangliosides present on the surface of rodent mast cells in maintaining cell structure was examined using RBL-2H3 mast cells and two mutant cell lines (E5 and D1) deficient in the gangliosides, GM1 and the α-galactosyl derivatives of the ganglioside GD1b. The two deficient cell lines were morphologically different from each other as well as from the parental RBL-2H3 cells. Actin filaments in RBL-2H3 and E5 cells were under the plasma membrane following the spindle shape of the cells, whereas in D1 cells, they were concentrated in large membrane ruffles. Microtubules in RBL-2H3 and E5 cells radiated from the centrosome and were organized into long, straight bundles. The bundles in D1 cells were thicker and organized circumferentially under the plasma membrane. The endoplasmic reticulum, the Golgi complex, and the secretory granule matrix were also altered in the mutant cell lines. These results suggest that the mast cell- specific α-galactosyl derivatives of ganglioside GD1b and GM1 are important in maintaining normal cell morphology.
BadizadeganKWheelerHEFujinagaYLencerWI (2004) Trafficking of cholera toxin-ganglioside GM1 complex into Golgi and induction of toxicity depend on actin cytoskeleton. Am J Physiol Cell Physiol, 287:C1453–1462.
3.
BarsumianELIserskyCPetrinoMGSiraganianRP (1981) IgE-induced histamine release from rat basophilic leukemia cell lines: isolation of releasing and nonreleasing clones. Eur J Immunol, 11: 317–323.
ChengPCCherukuriADystraMMalapatiSSproulTChenMRPierceSK (2001) Floating the raft hypothesis: the roles of lipid rafts in B cell antigen receptor function. Semin Immunol, 13: 107–114.
9.
ChichiliGRRodgersW (2007) Clustering of membrane raft proteins by the actin cytoskeleton. J Biol Chem, 282:36682–36691.
10.
ChichiliGRRodgersW (2009) Cytoskeleton-membrane interactions in membrane raft structure. Cell Mol Life Sci, 66:2319–2328.
11.
CondeCCáceresA (2009) Microtubule assembly, organization and dynamics in axons and dendrites. Nat Rev Neurosci, 10: 319–332.
12.
CrespoPMZuritaARGiraudoCGMaccioniJFDaniottiJL (2004) Ganglioside glycosyltransferases and newly synthesized gangliosides are excluded from detergent-insoluble complexes of Golgi membranes. Biochem J, 377:561–568.
13.
de GraffenriedCLBertozziCR (2004) The roles of enzyme localization and complex formation in glycan assembly within the Golgi apparatus. Curr Opin Cell Biol, 16:356–363.
FieldKAHolowkaDBairdB (1995) FcyRI-mediated recruitment of p53/56Lyn to detergent-resistant membrane domains accompanies cellular signaling. Proc Natl Acad Sci USA, 92:9201–9205.
16.
FrigeriLApgarJR (1999) The role of actin microfilaments in the down-regulation of the degranulation response in RBL-2H3 mast cells. J Immunol, 162:2243–2250.
17.
FujitaAChengJFujimotoT (2009) Segregation of GM1 and GM3 clusters in the cell membrane depends on the intact actin cytoskeleton. Biochim Biophys Acta, 1791:388–396.
18.
GillardBKThurmonLTHarrelRGCapetanakiYSaitoMYuRKMarcusDM (1994) Biosynthesis of glycosphingolipids is reduced in the absence of vimentin intermediate filament network. J Cell Sci, 107:3545–3555.
19.
GorsiBStringerSE (2007) Tinkering with heparan sulfate sulfation to steer development. Trends Cell Biol, 17:173–177.
20.
GuoNHerGRReinholdVNBrennamMJSiraganianRPGinsburgV (1989) Monoclonal antibody AA4 which inhibits binding of IgE to high affinity receptors on rat basophilic leukemia cells, binds to novel α-galactosyl derivates of ganglioside GD1b. J Biol Chem, 264:13267–13272.
21.
GuptaNDeFrancoAL (2003) Visualizing lipid raft dynamics and early signaling events during antigen receptor-mediated B-lymphocyte activation. Mol Biol Cell, 14:432–444.
22.
HakomoriS (1981) Glycosphingolipids in cellular interaction, differentiation, and oncogenesis. Annu Rev Biochem, 50:733–764.
23.
HakomoriS (1996) Tumor malignancy defined by aberrant glycosylation and sphingo(glyco)lipid metabolism. Cancer Res, 56: 5309–5318.
24.
HakomoriS (2000) Traveling for the glycosphingolipid path. Glycoconj J, 17:627–647.
25.
HatayamaTTsujiokaKWakatsukiTKitamuraTImaharaH (1992) Effects of low culture temperature on the induction of hsp70 mRNA and the accumulation of hsp70 and hsp105 in mouse FM3A cells. J Biochem, 111:484–490.
26.
HolowkaDBairdB (2001) FcyRI as a paradigm for a lipid raft-dependent receptor in hematopoietic cells. Semin Immunol, 13: 99–105.
27.
HolowkaDSheetsEDBairdB (2000) Interactions between FcyRI and lipid raft components are regulated by the actin cytoskeleton. J Cell Sci, 113:1009–1019.
28.
IkamiTIshidaHKisoM (2000) Synthesis and biological activity of glycolipids, with a focus on gangliosides and sulfatide analogs. Methods Enzymol, 311:547–568.
29.
IlschnerSBrandtR (1996) The transition of microglia to a ramified phenotype is associated with the formation of stable acetylated and detyrosinated microtubules. Glia, 18:129–140.
30.
JordanSRodgersW (2003) T cell glycolipid-enriched membrane domains are constitutively assembled as membrane patches that translocate to immune synapses. J Immunol, 171:78–87.
31.
JouILeeJHParkSYYoonHJJoeEHParkEJ (2006) Gangliosides trigger inflammatory responses via TLR4 in brain glia. Am J Pathol, 168:1619–1630.
32.
KarlssonKA (1989) Animal glycosphingolipids as membrane attachment sites for bacteria. Annu Rev Biochem, 58:309–350.
33.
KatoNNakanishiMHirashimaN (2003) Cholesterol depletion inhibits store-operated calcium currents and exocytotic membrane fusion in RBL-2H3 cells. Biochemistry, 42:11808–11814.
KuznetsovSALangfordGMWeissDG (1992) Actin-dependent organele movement in squid axoplasm. Nature, 356:722–725.
36.
KwikJBoyleSFooksmanDMargolisLSheetzMPEdidinM (2003) Membrane cholesterol, lateral mobility, and the phosphatidylinositol 4,5-biphosphate-dependent organization of cell actin. Proc Natl Acad Sci USA, 100:13964–13969.
37.
LennePFWawrezinieckLConchonaudFWurtzOBonedAGuoXJRigneaultH (2006) Dynamic molecular confinement in the plasma membrane by microdomains and the cytoskeleton meshwork. EMBO J, 25:3245–3256.
38.
LevitanIGoochK (2007) Lipid rafts in membrane-cytoskeleton interactions and control of cellular biomechanics: actions of oxLDL. Antioxid Redox Signal, 9:1519–1534.
39.
LinJShawAS (2005) Getting downstream without a raft. Cell, 121: 815–816.
40.
Meivar-LevyISabanayHBershadskyADFutermanAH (1997) The role of sphingolipids in the maintenance of fibroblast morphology. J Biol Chem, 17:1558–1564.
41.
MiceliMCMoranMChungCDPatelVPLowTZinnantiW (2001) Co-stimulation and counter-stimulation: lipid raft clustering controls TCR signaling and functional outcomes. Semin Immunol, 13:115–128.
42.
MitchellJSBrownWSWoodsideDGVanderslicePMcIntyreBW (2009) Clustering T-cell GM1 lipid rafts increases cellular resistance to shear on fibronectin through changes in integrin affinity and cytoskeletal dynamics. Immunol Cell Biol, 87:324–336.
43.
MoranMMiceliMC (1998) Engagement of GPI-linked CD48 contributes to TCR signals and cytoskeletal reorganization: a role for lipid rafts in T cell activation. Immunity, 9:787–796.
44.
NaderHBBuonassisiVColburnPDietrichCP (1989) Heparin stimulates the synthesis and modifies the sulfation pattern of heparan sulfate proteoglycan from endothelial cells. J Cell Physiol, 140:305–310.
45.
NakamuraNLoweMLevineTPRabouilleCWarrenG (1997) The vesicle docking protein p115 binds GM130, a cis-Golgi matrix protein, in a mitotically regulated manner. Cell, 89:445–455.
46.
NeblTPestonjamaspKNLeszykJDCrowlwyJLOhSWLunaEJ (2002) Proteomic analysis of a detergent-resistant membrane skeleton from neutrophil plasma membranes. J Biol Chem, 277:43399–43409.
47.
NgMMChangFBurgessDR (2005) Movement of membrane domains and requirement of membrane signaling molecules for cytokinesis. Dev Cell, 9:781–790.
48.
OakleyCEOakleyBR (1989) Identification of gamma-tubulin, a new member of the tubulin superfamily encoded by mipA gene of Aspergillus nidulans. Nature, 338:662–664.
49.
OliverCSaharaNKitaniSRobbinsARMertzLMSiraganianRP (1992) Binding of monoclonal antybody AA4 to gangliosides on rat basophilic leukemia cells produces changes similar to those seen with Fc epsilon receptor activation. J Cell Biol, 116:635–646.
50.
PalestiniPPittoMTedeschiGFerrarettoAParentiMBrunnerJMasseriniM (2000) Tubulin anchoring to glycolipid-enriched, detergent-resistant domains of the neuronal plasma membrane. J Biol Chem, 276:9978–9985.
51.
ParkJYKimHYJouIParkSM (2008) GM1 induces p38 and microtubule dependent ramification of rat primary microglia in vitro. Brain Res, 1244:13–23.
52.
PfeifferJRSeagraveJCDavisBHDeaninGGOliverJM (1985) Membrane and cytoskeletal changes associated with IgE-mediated serotonin release from rat basophilic leukemia cells. J Cell Biol, 101:2145–2155.
53.
PikeLJ (2003) Lipid rafts: bringing order to chaos. J Lipid Res, 44:655–667.
54.
PikeLJ (2009) The challenge of lipid rafts. J Lipid Res, 50(suppl): 323–328.
55.
ReynoldsES (1963) The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J Cell Biol, 17:208–212.
56.
RiveroSCardenasJBornensMRiosRM (2009) Microtubule nucleation at cis-side of the Golgi apparatus requires AKAP450 and GM130. EMBO J, 28:1016–1028.
57.
RodgersWGlaserM (1993) Distributions of proteins and lipids in the erythrocyte membrane. Biochemistry, 32:12591–12598.
58.
SaharaNSiraganianRPOliverC (1990) Morphological changes induced by the calcium ionophore A23187 in rat basophilic leukemia (2H3) cells. J Histochem Cytochem, 38:975–983.
59.
SheetsEDHolowkaDBairdB (1999) Membrane organization in immunoglobulin E receptor signaling. Curr Opin Chem Biol, 3: 95–99.
60.
SilbertJESugumaranG (2002) Biosynthesis of chondroitin/dermatan sulfate. IUBMB Life, 54:177–186.
61.
Silveira e SouzaAMMMazucatoVMde CastroROMatioliFCiancagliniPPaulinoTPJamurMC (2008) The alpha-galactosyl derivatives of ganglioside GD1b are essential for the organization of lipid rafts in RBL-2H3 mast cells. Exp Cell Res, 13:2515–2528.
62.
SimonsKToomreD (2000) Lipid rafts and signal transduction. Nat Rev Mol Cell Biol, 1:31–39.
63.
SonninoSMauriLChigornoVPrinettiA (2007) Gangliosides as components of lipid membrane domains. Glycobiology, 17: 1R–13R.
64.
SperoDABrowningETRoisenFJ (1986) Modification of micro-tubule-microfilament interactions during neurite outgrowth. Ann N Y Acad Sci, 466:849–853.
65.
SperoDARoisenFJ (1984) Ganglioside-mediated enhancement of the cytoskeletal organization and activity in neuro-2a neuroblastoma cells. Brain Res, 315:37–48.
66.
StephanVSeibtADukanovicDSkasaMSwaimWDBerensteinEHSiraganianRP (1997) Anti-ganglioside monoclonal antibody AA4 selectively inhibits IgE-induced signal transduction pathways in rat basophilic leukemia cells. Mol Immunol, 34:227–235.
67.
StrackeMLBascianoLKSiraganianRP (1987) Binding properties and histamine release in variants of rat basophilic leukemia cells with changes in the IgE receptor. Immunol Lett, 14:287–292.
68.
SwaimWDMinoguchiKOliverCHamawyMMKiharaHStephanVBerensteinEH (1994) The anti-ganglioside monoclonal antibody AA4 induces protein tyrosine phosphorylations, but not degranulation, in rat basophilic leukemia cells. J Biol Chem, 269: 19466–19473.
69.
ThybergJMoskalewskiS (1999) Role of microtubules in the organization of the Golgi complex. Exp Cell Res, 246:263–279.
70.
TrindadeESOliverCJamurMCFrancoCRCRochaHAOBoucasRIJarrougeTR (2008) The binding of heparin to the extracellular matrix of endothelial cells up-regulates the synthesis of an antithrombotic heparan sulfate proteoglycan. J Cell Physiol, 217:328–337.
71.
ValensinSPaccaniSRUlivieriCMercatiDPaciniSPatrussiLHirstT (2002) F-actin dynamics control segregation of the TCR signaling cascade to clustered lipid rafts. Eur J Immunol, 32: 435–444.
72.
WolfAAJoblingMGSaslowskyDEKernEDrakeKRKenworthyAKHolmesRK (2008) Attenuated endocytosis and toxicity of a mutant cholera toxin with decreased ability to cluster ganglioside GM1 molecules. Infect Immun, 76:1476–1484.
73.
XavierRBrennanTLiQMcCormackCSeedB (1998) Membrane compartmentation is required for efficient T cell activation. Immunity, 8:723–732.
74.
YamashitaTWadaRSasakiTDengCBierfreundUSandhoffKProiaRL (1999) A vital role for glycosphingolipid synthesis during development and differentiation. Proc Natl Acad Sci USA, 96:9142–9147.
75.
YatesAJWarnerJKStockSMMcQuarrieIG (1989) Ganglioside synthesis and transport in regenerating sensory neurons of the rat sciatic nerve. Brain Res, 479:277–282.
