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Expression of neutral glycosphingolipids and gangliosides in human skeletal and heart muscle determined by indirect immunofluorescence staining

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Abstract

The expression of neutral glycosphingolipids and gangliosides has been studied in human skeletal and heart muscle using indirect immunofluorescence microscopy. Transversal and longitudinal cryosections were immunostained with specific monoclonal and polyclonal antibodies against the neutral glycosphingolipids lactosylceramide, globoside, Forssman glycosphingolipid, gangliotetraosylceramide, lacto-N-neotetraosylceramide and against the gangliosides GM3(Neu5Ac) and GM1(Neu5Ac). To confirm the lipid nature of positive staining, control sections were treated with methanol and chloroform:methanol (1:1) before immunostaining. These controls were found to be either negative or strongly reduced in fluorescence intensity, suggesting that lipid bound oligosaccharides were detected. In human skeletal muscle, lactosylceramide was found to be the main neutral glycosphinogolipid. Globoside was moderately expressed, lacto-N-neotetraosylceramide and gangliotetraosylceramide were minimally expressed and Forssman glycosphingolipid was not detected in human skeletal muscle. The intensities of the immunohistological stains of GM3 and GM1 correlated to the fact that GM3 is the major ganglioside in skeletal muscle whereas GM1 is expressed only weakly. In human heart muscle globoside was the major neutral glycosphingolipid. Lactosylceramide and lacto-N-neotetraosylceramide were moderately expressed, gangliotetraosylceramide was weakly expressed and the Forssman glycosphingolipid was not expressed at all in cardiac muscle. GM3 and GM1 were detected with almost identical intensity. All glycosphingolipids were present in plasma membranes as well as at the intracellular level.

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Author information

Authors and Affiliations

  1. Department of Chemistry and Biochemistry, School of Medicine, University of Zagreb, Šalata 3b, 41000, Zagreb, Croatia

    Melita Čačić & Ivan Kračun

  2. Institute for Cell Culture Technology, University of Bielefeld, 33501, Bielefeld, Germany

    Johannes Müthing

  3. Clinic for Poultry of the Hannover School of Veterinary Medicine, 30559, Hannover, Germany

    Ulrich Neumann

  4. Institute for Developmental Biology, University of Bielefeld, 33501, Bielefeld, Germany

    Sabine Weber-Schürholz

Authors
  1. Melita Čačić
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  2. Johannes Müthing
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  3. Ivan Kračun
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  4. Ulrich Neumann
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  5. Sabine Weber-Schürholz
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Additional information

Abbreviations used: BSA, bovine serum albumin; DAPI, 4′,6-diamidine-2-phenylindole-dihydrochloride; DTAF, fluorescein isothiocyanate derivative; GSL(s), glycosphingolipid(s); Neu5Ac,N-acetylneuraminic acid [50]; PBS, phosphate buffered saline. The designation of the following glycosphingolipids follows the IUPAC-IUB recommendations [51] and the nomenclature of Svennerholm [52]. Lactosylceramide or LacCer, Galβ1-4Glcβ1-1Cer; gangliotriaosylceramide or GgOse3Cer, GalNAcβ1-4Galβ1-4Glcβ1-1Cer; globotriaosylceramide or GbOse3Cer, Galαl-4Galβl-4Glcβl-1Cer; gangliotetraosylceramide or GgOse4Cer, Galβ1-3GalNAcβ1-4Galβ1-4Glcβ1-1Cer; globotetraosylceramide or GbOse4Cer, GalNAcβ1-3Galα1-4Galβ1-4Glcβ1-1Cer; lacto-N-neotetraosylceramide or nLcOse4Cer, Galβ1-4GlcNAcβ1-3Galβ1-4Glcβ1-1Cer; Forssman GSL or GbOse3Cer, GalNAcα1-3GalNAcβ1-3Galα1-4Galβ1-4Gleβ1-1Cer; GM3, II3Neu5Ac-LacCer; GM2, II3Neu5Ac-GgOse3Cer; GM1, II3Neu5Ac-GgOse4Cer; GD3 II3(Neu5Ac)2-LacCer; GD2, II3(Neu5Ac)2-GgOse3Cer; GD1a, IV3Neu5Ac, II3Neu5Ac-GgOse4Cer; GD1b, II3(Neu5Ac)2-GgOse4Cer.

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Čačić, M., Müthing, J., Kračun, I. et al. Expression of neutral glycosphingolipids and gangliosides in human skeletal and heart muscle determined by indirect immunofluorescence staining. Glycoconjugate J 11, 477–485 (1994). https://doi.org/10.1007/BF00731284

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  • DOI: https://doi.org/10.1007/BF00731284

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