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Fabry disease: renal sphingolipid distribution in the α-Gal A knockout mouse model by mass spectrometric and immunohistochemical imaging

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Abstract

Fabry disease is an X-linked lysosomal storage disease due to deficient α-galactosidase A (α-Gal A) activity and the resultant lysosomal accumulation of globotriaosylceramide (Gb3) and related lipids primarily in blood vessels, kidney, heart, and other organs. The renal distribution of stored glycolipid species in the α-Gal A knockout mouse model was compared to that in mice to assess relative distribution and absolute amounts of accumulated sphingolipid isoforms. Twenty isoforms of five sphingolipid groups were visualized by mass spectrometry imaging (MSI), and their distribution was compared with immunohistochemical (IHC) staining of Gb3, the major stored glycosphingolipid in consecutive tissue sections. Quantitative bulk lipid analysis of tissue sections was assessed by electrospray ionization with tandem mass spectrometry (ESI-MS/MS). In contrast to the findings in wild-type mice, all three analytical techniques (MSI, IHC, and ESI-MS/MS) revealed increases in Gb3 isoforms and ceramide dihexosides (composed mostly of galabiosylceramides), respectively. To our knowledge, this is the first report of the distribution of individual molecular species of Gb3 and galabiosylceramides in kidney sections in Fabry disease mouse. In addition, the spatial distribution of ceramides, ceramide monohexosides, and sphingomyelin forms in renal tissue is presented and discussed in the context of their biosynthesis.

Immunohistochemical images of a wild type (left) and Fabry mouse kidney (right)

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Authors’ contributions

The manuscript was written through contributions of all authors. All authors have given their approval to the final version of the manuscript. RJD generated the FKO mouse. RD participated in mouse colony management, sample collection, preparation, and coordination of the study. LKr and HF were responsible for the MSI data acquisition; LKu collected and evaluated the quantitative MS data. JR and BA participated in preparation of lipid samples and protein analysis. MS and MV analyzed and evaluated the MSI data. HH was responsible for the IHC and in situ studies. LKry performed the immunohistochemical analyses. JL, LKu, HH, KL, RJD, and VH wrote the paper.

Funding sources

The authors acknowledge the major direct support from the Ministry of Education, Youth and Sports of the Czech Republic (COST-CZ-LD13038, PRVOUK-P24/LF1/3, COST-CZ-LD13005 and UNCE 204011), Ministry of Health of the Czech Republic (Grant IGA MZ NT14015-3/2013), and Czech Science Foundation (P206/12/1150). Access to instrumental and other facilities was also supported by EU (COST BM1104, Operational Program Prague—Competitiveness project CZ.2.16/3.1.00/24023) and IMIC institutional research concept RVO61388971.

Conflict of interest

RJD serves as a consultant to Amicus Therapeutics and Genzyme Corp, holds shares of Amicus Therapeutics, receives grants from Genzyme Corp., and receives royalties from Genzyme Corp. and Shire HGT.

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Authors and Affiliations

  1. Institute of Inherited Metabolic Disorders, Charles University in Prague, Ke Karlovu 2, 128 08, Prague 2, Czech Republic

    Ladislav Kuchar, Robert Dobrovolny, Helena Hulkova, Jana Ledvinova, Lenka Kryspinova, Befekadu Asfaw & Jitka Rybová

  2. Institute of Microbiology, ASCR, v.v.i, Videnska 1083, 14220, Prague 4, Czech Republic

    Helena Faltyskova, Lukas Krasny, Michael Volny, Martin Strohalm, Karel Lemr & Vladimir Havlicek

  3. Institute of Chemical Technology in Prague, Technicka 5, 16228, Prague 6, Czech Republic

    Lukas Krasny

  4. Regional Centre of Advanced Technologies and Materials, Department of Analytical Chemistry, Faculty of Science, Palacky University, 17.listopadu 12, 771 46, Olomouc, Czech Republic

    Karel Lemr & Vladimir Havlicek

  5. Department of Genetics and Genomics Sciences, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA

    Robert J. Desnick

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  1. Ladislav Kuchar
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Corresponding author

Correspondence to Vladimir Havlicek.

Additional information

Published in the topical collection Mass Spectrometry Imaging with guest editors Andreas Römpp and Uwe Karst.

Ladislav Kuchar and Helena Faltyskova contributed equally to this work.

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Kuchar, L., Faltyskova, H., Krasny, L. et al. Fabry disease: renal sphingolipid distribution in the α-Gal A knockout mouse model by mass spectrometric and immunohistochemical imaging. Anal Bioanal Chem 407, 2283–2291 (2015). https://doi.org/10.1007/s00216-014-8402-7

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  • DOI: https://doi.org/10.1007/s00216-014-8402-7

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