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Selective Extraction and Effective Separation of Galactosylsphingosine (Psychosine) and Glucosylsphingosine from Other Glycosphingolipids in Pathological Tissue Samples

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Abstract

To facilitate the study of the chemical pathology of galactosylsphingosine (psychosine, GalSph) in Krabbe disease and glucosylsphingosine (GlcSph) in Gaucher disease, we have devised a facile method for the effective separation of these two glycosylsphingosines from other glycosphingolipids (GSLs) in Krabbe brain and Gaucher spleen samples. The procedure involves the use of acetone to selectively extract GalSph and GlcSph, respectively, from Krabbe brain and Gaucher spleen samples. Since acetone does not extract other GSLs except modest amounts of galactosylceramide, sulfatide, and glucosylceramide, the positively charged GalSph or GlcSph in the acetone extract can be readily separated from other GSLs by batchwise cation-exchange chromatography using a Waters Accell Plus CM Cartridge. GalSph or GlcSph enriched by this simple procedure can be readily analyzed by thin-layer chromatography or high-performance liquid chromatography.

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Acknowledgments

This study was supported by NIH grant R01 NS09626 (to YT Li), NIH grant RR02512 (to ME Haskins), NIH grant NCRR R24RR022826 and Louisiana Gene Therapy Research Consortium Tulane University (to B. Bunnell). The human brain samples from Krabbe patients and the age matched control subjects were obtained from the NICHD Brain and Tissue Bank for Developmental Disorders at the University of Maryland, Baltimore, MD. MALDI-MS analyses were performed at the NRPGM Core Facilities for Proteomics and Glycomics, Institute of Biological Chemistry, Academia Sinica, Taiwan, supported by an NSC grant NSC98-3112-B-001-023.

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Author notes

  1. Wayne R. Buck

    Present address: Cellular Molecular & Exploratory Toxicology, Abbott Laboratories, Abbott Park, North Chicago, IL, USA

Authors and Affiliations

  1. Department of Biochemistry, Tulane University School of Medicine, 1430 Tulane Avenue, SL-43, New Orleans, LA, 70112, USA

    Yu-Teh Li & Su-Chen Li

  2. Department of Pathology, Tulane National Primate Research Center, Covington, LA, USA

    Wayne R. Buck

  3. Department of Pathobiology, University of Pennsylvania School of Veterinary Medicine, Philadelphia, PA, USA

    Mark E. Haskins

  4. Institute of Biological Chemistry, Academia Sinica, Taipei, Taiwan

    Sz-Wei Wu & Kay-Hooi Khoo

  5. Section on Molecular Neurogenetics, Medical Genetics Branch, National Genome Research Institute, National Institute of Health, Bethesda, MD, USA

    Ellen Sidransky

  6. Center for Gene Therapy, Tulane University School of Medicine, New Orleans, LA, USA

    Bruce A. Bunnell

  7. Division of Gene Therapy, Tulane National Primate Research Center, Covington, LA, USA

    Bruce A. Bunnell

  8. Department of Pharmacology, Tulane University School of Medicine, New Orleans, LA, USA

    Bruce A. Bunnell

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  1. Yu-Teh Li
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  2. Su-Chen Li
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  7. Ellen Sidransky
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  8. Bruce A. Bunnell
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Correspondence to Yu-Teh Li.

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Special Issue: In honor of Dr. Robert Yu.

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Li, YT., Li, SC., Buck, W.R. et al. Selective Extraction and Effective Separation of Galactosylsphingosine (Psychosine) and Glucosylsphingosine from Other Glycosphingolipids in Pathological Tissue Samples. Neurochem Res 36, 1612–1622 (2011). https://doi.org/10.1007/s11064-010-0348-3

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  • DOI: https://doi.org/10.1007/s11064-010-0348-3

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