Abstract
The two clonal murine muscle cell lines G7 and G8, originally derived from the M114 line [20], represent unique models for comparative studies of myogenesis. Glycolipid synthesis was examined during differentiation using [3H]-galactose and [3H]-glucosamine as precursors. Upon G7 contact glucosylceramide labeling increased and nLcOse5Cer labeling stopped. During membrane fusion, glucosylceramide labeling stopped and lactosylceramide became the major synthetic product. G8 cells presented a different pattern, with increased labeling of GbOse3Cer during myogenesis. The major ganglioside synthesized by both myoblasts was GM3, and more complex structures were observed following completion of myotube formation. Total glycopeptide labeling increased when G8 myoblasts fused and remained elevated in myotubes, whereas no differences during fusion of G7 cells were noted. Upon comparison of the two clonal lines, the only consistent observation was a significant increase in the synthesis of total gangliosides and neutral glycolipid during cell contact and membrane fusion (p < 0.02). The results suggest that changes in the synthesis of specific glycolipid structures during myogenesis are unique to each muscle cell line examined. However, transient increases in synthesis of total myoblast gangliosides and neutral glycolipids may be a more general phenomenon, possibly by curbing proliferation or by altering myoblast membrane fluidity characteristics during differentiation.
Similar content being viewed by others
Abbreviations
- MG6:
-
VI3NeuAc-V4Gal-IV3GlcNAc-nLcOse4Cer
- TLC:
-
thin-layer chromatography
- HPTLC:
-
high performance thin-layer chromatography
- Gal:
-
galactose
- GlcNH:
-
glucosamine
- PBS:
-
phosphate buffered saline
- CK:
-
creatine kinase
References
Barbosa MLF, Pinto da Silva P: Restriction of glycolipids to the outer half of a plasma membrane: Concanavalin A labeling of membrane halves in Acanthamoeba castellanii. Cell 33: 959–966, 1983
Hakomori S-I: Glycosphingolipids in cellular interactions, differentiation and oncogenesis. Ann Rev Biochem 50: 733–764, 1981
Chien J-L, Hogan EL: Glycosphingolipids of skeletal muscle. ACS Symp Ser 128: 135–148, 1980
Chien J-L, Hogan EL: Novel pentahexosyl ganglioside of the globo series purified from chicken muscle. J Biol Chem 258: 10727–10730, 1983
Nakamura K, Nagashima M, Sekine M, Igarashi M, Ariga T, Atsumi T, Miyatake T, Suzuki A, Yamakawa T: Gangliosides of hog skeletal muscle. Biochim Biophys Acta 752: 291–300, 1983
Svennerholm L, Bruce A, Mansson J-E, Rynmark B-M, Vanier M-T: Sphingolipids of human skeletal muscle. Biochim Biophys Acta 280: 626–636, 1972
Saito M, Rosenberg A: Glycolipids and their developmental patterns in chick thigh and leg muscle. J. Lipid Res 23: 3–8, 1982
Clark GF, Smith PB: Studies on glycoconjugate metabolism in developing skeletal muscle membranes. Biochim Biophys Acta 755: 5664–64, 1983
Max SR, Nelson PG, Brady RO: The effect of denervation on the composition or muscle gangliosides. J. Neurochem 17: 1517–1520, 1970
Higatsberger MR, Auff E: Gangliosides in rabbit and human skeletal muscle with denervation atrophy. J. Neurol 231:79–82, 1984
Inestrosa NC: Differentiation of skeletal muscle cells in culture. Cell Struct Func 7: 91–109, 1982
Wakelam MJO: The fusion of myoblasts. Biochem J 228: 1–12, 1985
Allen RE: Muscle cell culture as a tool in animal growth research. Fed Proc 46: 290–294, 1987
Yasin R. Walsh FS, Landon DN, Thompson EJ: New approaches to the study of human dystrophic muscle cells in culture. J Neurol Sci 58: 315–334, 1983
Dubois C, Hauttecoeur B, Coulon-Morclec M-J, Montarras D, Rampini C, Fiszman MY: Changes in ganglioside metabolism during in vitro differentiation of quail embryo myoblasts. Dev Biol 105: 509–517, 1984
McEvoy FA, Ellis DE: Glycolipids and myoblast differentiation. Biochem Soc Trans 5: 1719–1721, 1977
Whatley R, Ng SK-C, Rogers J, McMurray WC, Sanwal BD: Developmental changes in gangliosides during myogenesis of a rat myoblast cell line and its resistant varients. Biochem Biophys Res Comm 70: 180–185, 1976
Leskawa KC, Erwin RE, Buse PE, Hogan EL: Glycosphingolipid biosynthesis during myogenesis of rat L6 cells in vitro. Molec Cell Biochem 83: 47–54, 1988
Nakamura K, Ariga T, Yahagi T, Miyatake Y, Suzuki A, Yamakawa T: Interspecies comparison of muscle gangliosides by two-dimensional thin-layer chromatography. J Biochem 94: 1359–1365, 1983
Christian CN, Nelson PG, Peacock J, Nirenberg M: Synapse formation between two clonal cell lines. Science 196: 995–998, 1977
Tettamanti G, Bonali F, Marchesini S, Zambotti V: A new procedure for the extraction, purification and fractionation of brain gangliosides. Biochim Biophys Acta 296: 160–170, 1973
Leskawa KC, Buse PE, Hogan EL, Garvin AJ: Glycosphingolipids of an alveolar rhabdomyosarcoma: a case study. Neurochem Path 2: 19–29, 1984
Williams MA, McCluer RH: The use of Sep-Pak C18 cartridges during the isolation of gangliosides. J Neurochem 35:266–269, 1980
Ledeen RW, Yu RK: Gangliosides: structure, isolation and analysis. Meth Enzymol 83: 139–191, 1982
Laskey RA, Mills AD: Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem 56: 335–341, 1975
Yates AJ, Thompson D: An improved assay of gangliosides separated by thin-layer chromatography. J Lipid Res 18: 660–663, 1979
Bailey RW, Bourne EI: Color reactions given by sugars and diphenylamine-aniline spray reagents on paper chromatograms. J Chromatog 4: 206–213, 1960
Leskawa KC, Dasgupta SV, Chien J-L, Hogan EL: A simplified procedure for the preparation of high specific activity tritiated GMl ganglioside and other glycosphingolipids. Analyt Biochem 140: 172–177, 1984
Krusius T, Finne J, Karrainen J, Jarnefelt J: Neutral and acidic glycopeptides in adult and developing rat brain. Biochim Biophys Acta 365: 80–92, 1974
Margolis RU: Acid mucopolysaccharides and properties of bovine whole brain, white matter and myelin. Biochim Biophys Acta 141: 91–102, 1967
Schauer R: Characterization of sialic acids. Meth Enzymol 50:64–89, 1978
Leskawa KC, Agranoff BW: Properties of an endogenous, membrane associated sialidase activity (N-acetylneuraminidase) of the goldfish visual system. Neurochem Res 8: 97–109, 1983
Konat G, Hogan EL, Leskawa KC, Gaunt G, Singh I: Abnormal glycosylation of myelin-associated glycoprotein in quaking mouse brain. Neurochem Int 10: 555–558, 1987
Ghebregzabher M, Rufini S, Monalsi B, Lato M: Thin-layer chromatography of carbohydrates. J Chromatog 127: 133–162, 1976
Kohama K, Ozawa E: A statistical method to compare the degree of muscle cell multiplication in different culture dishes. Develop Growth Differen 19: 139–148, 1977
Hughes BP: A method for the estimation of serum creatine kinase and its use in comparing creatine kinase and aldolase activity in normal and pathologic sera. Clin Chem Acta 7: 597–603, 1962
Peterson GL: A simplification of the protein assay method of Lowry et al. which is more generally applicable. Analyt Biochem 83: 346–356, 1977
Turner DC, Maier V, Eppenberger HM: Creatine kinase and aldolase isozyme transitions in cultures of chick skeletal muscle cells. Dev Biol 37: 63–89, 1974
Shainberg A, Yagil G, Yaffe D: Control of myogenesis in vitro by Ca++ concentration in nutritional medium. Exp Cell Res. 58: 163–167, 1970
Den H, Malinzak DA, Keating HJ, Rosenberg A: Influence of concanavalin A, wheat germ agglutinin and soybean agglutinin on the fusion of myoblasts in vitro. J Cell Biol 67: 826–834, 1975
Sandra A, Leon MA, Przybylski RJ: Suppresion of myoblast fusion by concanavalin A: possible involvement of membrane fluidity. J Cell Sci 28: 251–263, 1977
Gartner TK, Podleski TR: Evidence that a membrane bound lectin mediates the fusion of L6 myoblasts. Biochem Biophys Res Comm 70: 1142–1149, 1975
Gilfix BM, Sanwal BD: Inhibition of myoblast fusion by tunicamycin and pantomycin. Biochem Biophys Res Comm 96: 1184–1191, 1980
Olden K, Bernard BA, White SL, Parent JB: Function of the carbohydrate moieties of glycoproteins. J Cell Biochem 18:313–335, 1982
Cates GA, Kaur H, Sanwal BD: Inhibition of fusion of skeletal myoblasts by tunicamycin and its reversal by N-acetylglucosamine. Can J Biochem 62: 28–35, 1984
Spearman MA, Jamieson JC, Wright JA: Studies on the effect of glycoprotein processing inhibitors on fusion of L6 myoblast cell lines. Exp Cell Res 168: 116–126, 1987
Den H: Effect of monensin on myoblast fusion. Biochem Biophys Res Comm 126: 313–319, 1985
Yusef HKM, Pohlentz G, Sandhoff K: Tunicamycin inhibits ganglioside biosynthesis in rat liver Golgi apparatus by blocking sugar nucleotide transport across the membrane vesicles. Proc Natl Acad Sci, USA 80: 7075–7079, 1983
Guarnaccia SP, Shaper JH, Schnaar RL: Tunicamycin inhibits ganglioside biosynthesis in neuronal cells. Proc Natl Acad Sci, USA 80: 1551–1555, 1983
Saito M, Rosenberg A: Action of monensin, a monovalent cationophore, on cultured human fibroblasts: evidence that it induces high cellular accumulation of glucosyl- and lacto sylceramide (gluco- and lactocerebroside). Biochem 23: 1043–1046, 1984
Leskawa KC, Erwin RE, Hogan EL: Phospholipid biosynthesis during normal and dystrophic avian muscle cell differentiation. Life Sci 38: 147–153, 1986
Wakelam MJO: Stimulated inositol phospholipid breakdown and myoblast fusion. Biochem Soc Trans 14: 253–256, 1986
Elson HF, Yguerabide J: Membrane dynamics of differentiating cultured embryonic chick skeletal muscle cells by fluorescent microscopy techniques. J Supramol Struct 12: 47–61, 1979
Prives J, Shinitzky M: Increased membrane fluidity precedes fusion of muscle cells. Nature 268: 761–763, 1977
Herman BA, Fernandez SM: Changes in membrane dynamics associated with myogenic cell fusion. J Cell Physiol 94:253–264, 1978
Lee PM, Ketis NV, Barber KR, Grant CMW: Ganglioside headgroup dynamics. Biochim Biophys Acta 601: 302–314, 1980
Ollman M, Galla H-J: Ganglioside headgroups decrease lipid order in reconstituted phosphatidylcholine liposomes. FEBS Lett 179: 173–176, 1985
Thompson TE, Tillack TW: Organization of glycosphingolipids in bilayers and plasma membranes of mammalian cells. Ann Rev Biophys Chem 14: 361–386, 1985
Roisen FJ, Bird JWC, Ochoa GS, St.John AC, Yorke G: Ganglioside-enhanced myoblast fusion in vitro. J Cell Biol 95: 367A, 1982 (Abstract)
Yurchenco PD, Ceccarini C, Atkinson PH: Labeling complex carbohydrates of animal cells with monosaccharides. Meth Enzymol 50: 175–203, 1978
Usuki S, Hoops P, Sweeley CC: Growth control of human foreskin fibroblasts and identification of extracellular sialidase activity by 2-doxy-2, 3-dehydro-N-acetylneuraminic acid. J Biol Chem 263: 10595–10599, 1988
IUPAC-IUB: Eur J Biochem 79: 11–21, 1977
Svennerholm L: Ganglioside nomenclature. Adv Exp Med Biol 125: 11, 1980
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Leskawa, K.C., Hogan, E.L. Regulation of glycolipid synthesis during differentiation of clonal murine muscle cells. Mol Cell Biochem 96, 163–173 (1990). https://doi.org/10.1007/BF00420908
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00420908