Intact cell MALDI-TOF mass spectrometry on single bovine oocyte and follicular cells combined with top-down proteomics: A novel approach to characterise markers of oocyte maturation

Autor: Laura Bouguereau, Audrey Gargaros, Aurélie Marestaing, Valérie Labas, Ana-Paula Teixeira-Gomes, Lucie Spina, Svetlana Uzbekova
Přispěvatelé: Physiologie de la reproduction et des comportements [Nouzilly] (PRC), Centre National de la Recherche Scientifique (CNRS)-Université de Tours-Institut Français du Cheval et de l'Equitation [Saumur]-Institut National de la Recherche Agronomique (INRA), Plateforme d'Analyse Intégrative des Biomolécules, Institut National de la Recherche Agronomique (INRA), Infectiologie Animale et Santé Publique - IASP (Nouzilly, France), Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Recherche Agronomique (INRA), European Regional Development Fund (ERDF), Val-de-Loire Region general Committee, the French National Institute for Agricultural Research (INRA), French National Institute of Health and Medical Research (INSERM) under the SMHART (LTQ Velos Orbitrap high resolution mass spectrometer acquisition, no. 3069), InnovoMass (ICM-MS) projects, INRA, French National Research Agency., Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours-Centre National de la Recherche Scientifique (CNRS), Infectiologie et Santé Publique (UMR ISP), Institut National de la Recherche Agronomique (INRA)-Université de Tours, Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS), Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur] (IFCE)-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS), Institut National de la Recherche Agronomique (INRA)-Université de Tours (UT), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS)
Jazyk: angličtina
Rok vydání: 2017
Předmět:
0301 basic medicine
Proteomics
bovin
cumulus cells
mass spectrum analysis
Peptide
Meiotic maturation
MALDI-TOF mass spectrometry
Top-down proteomics
oocyte
bovine
01 natural sciences
Biochemistry
Ubiquitin
Ovarian Follicle
spectrométrie de masse
Follicular phase
Biologie de la reproduction
maturation de l'ovocyte
chemistry.chemical_classification
Reproductive Biology
ovocyte
Cell biology
Meiosis
medicine.anatomical_structure
Female
Single-Cell Analysis
follicule
Autre (Sciences du Vivant)
Proteases
[SDV.OT]Life Sciences [q-bio]/Other [q-bio.OT]
Biophysics
Biology
Mass spectrometry
reproduction
03 medical and health sciences
Histone H2B
medicine
Animals
Granulosa Cells
010401 analytical chemistry
[SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology
Oocyte
Molecular biology
0104 chemical sciences
030104 developmental biology
chemistry
Spectrometry
Mass
Matrix-Assisted Laser Desorption-Ionization
biology.protein
Oocytes
Cattle
Biomarkers
Zdroj: Journal of Proteomics
Journal of Proteomics, Elsevier, 2017, pp.1-54. ⟨10.1016/j.jprot.2017.03.027⟩
Journal of Proteomics, Elsevier, 2017, 175, pp.56-74. ⟨10.1016/j.jprot.2017.03.027⟩
Journal of Proteomics (175), 56-74. (2018)
Journal of Proteomics, 2017, 175, pp.56-74. ⟨10.1016/j.jprot.2017.03.027⟩
ISSN: 1874-3919
1876-7737
DOI: 10.1016/j.jprot.2017.03.027⟩
Popis: International audience; Intact cell MALDI-TOF mass spectrometry (ICM-MS) was adapted to bovine follicular cells from individual ovarian follicles to obtain the protein/peptide signatures (< 17 kDa) of single oocytes, cumulus cells (CC) and granulosa cells (GC), which shared a total of 439 peaks. By comparing the ICM-MS profiles of single oocytes with CC before and after in vitro maturation (IVM), 71 different peaks were characterised, and their relative abundance was found to vary depending on the stage of oocyte meiotic maturation. To identify these endogenous biomolecules, top-down workflow using high resolution MS/MS (TD HR-MS) was performed on the protein extracts from oocytes, CC and GC. The TD HR-MS proteomic approach allowed for: (1) identification of 386 peptide/proteoforms encoded by 194 genes; and (2) characterisation of proteolysis products likely resulting from the action of kallikreins and caspases. In total, 136 peaks observed by ICM-MS were annotated by TD HR-MS (ProteomeXchange PXD004892). Among these, 16 markers of maturation were identified, including IGF2 binding protein 3 and hemoglobin B in the oocyte, thymosins beta-4 and beta-10, histone H2B and ubiquitin in CC. The combination of ICM-MS and TD HR-MS proved to be a suitable strategy to identify non-invasive markers of oocyte quality using limited biological samples. Biological significance Intact cell MALDI-TOF mass spectrometry on single oocytes and their surrounding cumulus cells, coupled to an optimised top-down HR-MS proteomic approach on ovarian follicular cells, was used to identify specific markers of oocyte meiotic maturation represented by whole low molecular weight proteins or products of degradation by specific proteases.
Databáze: OpenAIRE
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