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Early stage fetal neocortex exhibits a complex ganglioside profile as revealed by high resolution tandem mass spectrometry

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Abstract

In this study we report on the first mass spectrometric (MS) investigation of gangliosides and preliminary assessment of the expression and structure in normal fetal neocortex in early developmental stages: 14th (Neo14) and 16th (Neo16) gestational weeks. Ganglioside analysis was carried out using a hybrid quadrupole time-of-flight (QTOF) MS with direct sample infusion by nanoelectrospray ionization (nanoESI) in the negative ion mode. Under optimized conditions a large number of glycoforms i.e. 75 in Neo14 and 71 in Neo16 mixtures were identified. The ganglioside species were found characterized by a high diversity of the ceramide constitution, an elevated sialylation degree (up to pentasialylated gangliosides-GP1) and sugar cores modified by fucosylation (Fuc) and acetylation (O-Ac). Direct comparison between Neo14 and Neo16 revealed a prominent expression of monosialylated structures in the Neo16 as well as the presence of a larger number of polysialylated species in Neo14 which constitutes a clear marker of rapid development-dependant changes in the sialylation. Also the MS screening results highlighted that presumably O-acetylation process occurs faster than fucosylation. CID MS/MS under variable collision energy applied for the first time for structural analysis of a fucosylated pentasialylated species induced an efficient fragmentation with generation of ions supporting Fuc-GP1d isomer in early stage fetal brain neocortex.

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Abbreviations

Ac:

Acetyl/acetylation

Cer:

Ceramide

gw:

Gestational weeks

CID:

Collision-induced dissociation

Neu5Ac:

N-acetyl neuraminic acid

NanoESI:

Nanoelectrospray ionization

MS:

Mass spectrometry

MS/MS:

Tandem mass spectrometry

TIC:

Total ion chromatogram

QTOF MS:

Quadrupole time-of-flight mass spectrometer/spectrometry

LM:

Low mass resolution

HM:

High mass resolution

LacCer:

Galβ4Glcβ1Cer

GM3:

II3-α-Neu5Ac-LacCer

GD3:

II3-α-(Neu5Ac)2-LacCer

GT3:

II3-α-(Neu5Ac)3-LacCer

GM2:

II3-α-Neu5Ac-Gg3Cer

GD2:

II3-α-(Neu5Ac)2-Gg3Cer

GM1a or GM1:

II3-α-Neu5Ac-Gg4Cer

GM1b:

IV3-α-Neu5Ac-Gg4Cer

GalNAc-GM1b:

IV3-α-Neu5Ac-Gg5Cer

GD1a:

IV3-α-Neu5Ac,II3-α-Neu5Ac-Gg4Cer

GD1b:

II3-α-(Neu5Ac)2-Gg4Cer

GT1b:

IV3-α-Neu5Ac,II3-α-(Neu5Ac)2-Gg4Cer

GQ1b:

IV3-α-(Neu5Ac)2,II3-α-(Neu5Ac)2-Gg4Cer

nLM1 or 3′-nLM1:

IV3-α-Neu5Ac-nLc4Cer

LM1 or 3′-isoLM1:

IV3-α-Neu5Ac-Lc4Cer

nLD1:

disialo-nLc4Cer

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Acknowledgements

This work was supported by the European Social Fund, through the project POSDRU 107/1.5/S/78702, EU FP7 MARIE CURIE-PIRSES-GA-2010-269256 and by the Romanian National Authority for Scientific Research through the projects PN-II-ID-PCE-2011-3-0047 and PN-II-PCCA-2011-142.

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Authors and Affiliations

  1. Department of Neonatology, University of Medicine and Pharmacy “Victor Babes”, Eftimie Murgu Square 2, 300041, Timisoara, Romania

    Roxana M. Ghiulai & Constantin Ilie

  2. Department of Chemistry and Biology, “Aurel Vlaicu” University, Revolutiei Blvd. 77, 310130, Arad, Romania

    Mirela Sarbu & Alina D. Zamfir

  3. Department of Physics, West University Timisoara, Vasile Parvan Blvd. 4, 300223, Timisoara, Romania

    Mirela Sarbu

  4. Department of Chemistry and Biochemistry, University of Zagreb Medical School, 10000, Zagreb, Croatia

    Željka Vukelić

  5. Mass Spectrometry Laboratory, National Institute for Research and Development in Electrochemistry and Condensed Matter, Plautius Andronescu 1, 300224, Timisoara, Romania

    Alina D. Zamfir

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  1. Roxana M. Ghiulai
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  2. Mirela Sarbu
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  3. Željka Vukelić
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  4. Constantin Ilie
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  5. Alina D. Zamfir
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Correspondence to Alina D. Zamfir.

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Roxana M. Ghiulai and Mirela Sarbu have equal contribution.

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Ghiulai, R.M., Sarbu, M., Vukelić, Ž. et al. Early stage fetal neocortex exhibits a complex ganglioside profile as revealed by high resolution tandem mass spectrometry. Glycoconj J 31, 231–245 (2014). https://doi.org/10.1007/s10719-014-9517-y

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