What is the clinical evidence from this study?

Study design: Other. Intervention: Two-stage MS/MS spectral matching method. Primary outcome: Identification and comparison of ganglioside species and lipid forms.

February 10, 2022Open Access

Analysis and Comparison of Mouse and Human Brain Gangliosides via Two-Stage Matching of MS/MS Spectra

FHFanran HuangUniversity of Florida LBLaura S. BaileyUniversity of Florida TGTianqi GaoChangchun University of Chinese Medicine

Key Result

A novel two-stage MS/MS spectral matching method successfully characterized endogenous gangliosides, revealing that mouse brains predominantly contain 36-carbon ceramides while human brains predominantly contain 38-carbon ceramides.

Structured PICO

Population

15 human postmortem brain samples and mouse brain tissues used to validate a new MS/MS spectral matching method for ganglioside analysis.

Intervention

Two-stage matching of MS/MS spectra (carbohydrate and glycolipid product ions) coupled with LC-MS/MS

Outcome

Identification and characterization of endogenous ganglioside species and their lipid formssurrogate

A novel two-stage MS/MS spectral matching method enables detailed profiling of ganglioside species and their lipid forms in complex biological tissues like human and mouse brains.

Limitations

Absolute GSL quantitation would not be accurate without labeled GSL internal standards.
Lack of an extensive database of reference MS/MS spectra for GSL identification.
Human brain samples were derived from the dorsolateral prefrontal cortex and may not reflect whole human brain expression levels.
Did not try to identify GT1b, another major brain ganglioside, as the library did not contain its reference MS/MS spectrum.
GSL epimers are still difficult to identify.

Abstract

Image: see text Glycosphingolipids (GSLs), including gangliosides, are essential components of the cell membrane. Because of their vital biological functions, a facile method for the analysis and comparison of GSLs in biological issues is desired. To this end, a new method for GSL analysis was developed based on two-stage matching of the carbohydrate and glycolipid product ions of experimental and reference MS/MS spectra of GSLs. The applicability of this method to the analysis of gangliosides in biological tissues was verified using human plasma and mouse brains spiked with standards. The method was then used to characterize endogenous gangliosides in mouse and human brains. It was shown that each endogenous ganglioside species had varied lipid forms and that mouse and human brains had different compositions of ganglioside species and lipid forms. Moreover, a 36-carbon ceramide is found to represent the major lipid form for mouse brain gangliosides, while the major lipid form for most human brain gangliosides is a 38-carbon ceramide. This study has verified that the two-stage MS/MS spectral matching method could be used to study gangliosides or GSLs and their lipid forms in complex biological samples, thereby having a broad application.

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