Quantification of galactosylsphingosine in the twitcher mouse using electrospray ionization-tandem mass spectrometry

Peter C Sharp,Phillip D Whitfield,Rosanne Taylor,Peter Meikle

doi:10.1016/s0022-2275(20)31539-x

Peter C Sharp, Phillip D Whitfield + Show 2 more

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https://doi.org/10.1016/s0022-2275(20)31539-x

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Journal: Journal of Lipid Research	Publication Date: Dec 1, 2001
Citations: 54	License type: cc-by

Affiliation: South Australia Pathology

Abstract

Globoid cell leukodystrophy (Krabbe disease) is an autosomal recessive inherited neurodegenerative disorder caused by the deficiency of the lysosomal enzyme beta-galactosylceramidase. The pathogenesis of the disorder has been proposed to arise from the accumulation of the cytotoxic metabolite galactosylsphingosine (psychosine). The twitcher mouse is a naturally occurring murine model of globoid cell leukodystrophy. We have developed a rapid, sensitive, and specific mass spectrometric method for determining the galactosylsphingosine concentration in the tissues of twitcher mice. Galactosylsphingosine is extracted from the tissues in methanol, isolated using strong cation-exchange and C18 solid-phase extraction chromatography, and then directly analyzed using electrospray ionization-tandem mass spectrometry. A lactosylsphingosine internal standard has been employed for quantification. The assay demonstrated significant accumulation of galactosylsphingosine in the brain, spinal cord, and kidney of twitcher mice. It is anticipated that this method may be of use in the monitoring of experimental therapies for globoid cell leukodystrophy.

Highlights

Globoid cell leukodystrophy (Krabbe disease) is an autosomal recessive disorder caused by the deficiency of the lysosomal enzyme ␤-galactosylceramidase (EC 3.2.1.46) [1]
Journal of Lipid Research Volume 42, 2001 the pathogenesis of the disease has been proposed to arise from the storage of a cytotoxic metabolite, galactosylsphingosine, which is a substrate for the missing enzyme [3]
Galactosylsphingosine and the internal standard (ISTD) were characterized by means of a precursor ion scan for m/z 282

Summary

Introduction

Globoid cell leukodystrophy (Krabbe disease) is an autosomal recessive inherited neurodegenerative disorder caused by the deficiency of the lysosomal enzyme ␤-galactosylceramidase. Globoid cell leukodystrophy (Krabbe disease) is an autosomal recessive disorder caused by the deficiency of the lysosomal enzyme ␤-galactosylceramidase (EC 3.2.1.46) [1]. In globoid cell leukodystrophy galactosylceramide, the primary natural substrate of the deficient enzyme does not accumulate in the tissues of affected individuals. We describe the development of a rapid, sensitive, and specific method using electrospray ionization-tandem mass spectrometry (ESI-MS/MS) to characterize and quantify underivatized galactosylsphingosine from the tissues of twitcher mice

Methods

Results

Conclusion