title: Analysis and characterisation of bovine oocyte and embryo biomarkers by matrix-assisted desorption ionisation mass spectrometry imaging.
abstract: In the field of 'single cell analysis', many classical strategies like immunofluorescence and electron microscopy are the primary techniques of choice. However, these methodologies are time consuming and do not permit direct identification of specific molecular classes, such as lipids. In the present study, a novel mass spectrometry-based analytical approach was applied to bovine oocytes and embryos. This new metabolomics-based application uses mass spectrometry imaging (MSI), efficient data processing and multivariate data analysis. Metabolic fingerprinting (MF) was applied to the analysis of unfertilised oocytes, 2-, 4- and 8-cell embryos and blastocysts. A semiquantitative strategy for sphingomyelin [SM (16:0)+Na](+) (m/z 725) and phosphatidylcholine [PC (32:0)+Na](+) (m/z 756) was developed, showing that lipid concentration was useful for selecting the best metabolic biomarkers. This study demonstrates that a combination of MF, MSI features and chemometric analysis can be applied to discriminate cell stages, characterising specific biomarkers and relating them to developmental pathways. This information furthers our understanding of fertilisation and preimplantation events during bovine embryo development.
authors.affiliation: Department of Animal Reproduction, College of Veterinary Medicine and Animal Science, São Paulo University, Av. Prof. Dr. Orlando Marques de Paiva, 87 - Cidade Universitária, 05508-270, São Paulo, SP, Brazil.
authors.forename: Roseli F
authors.initials: RF
authors.lastname: Gonçalves
authors.affiliation: Innovare Biomarkers Laboratory, Medicine and Experimental Surgery Nucleus, University of Campinas, Rua Cinco de Junho, 350 - Barão Geraldo, 13083-877, Campinas, SP, Brazil.
authors.forename: Mónica S
authors.initials: MS
authors.lastname: Ferreira
authors.affiliation: Innovare Biomarkers Laboratory, Medicine and Experimental Surgery Nucleus, University of Campinas, Rua Cinco de Junho, 350 - Barão Geraldo, 13083-877, Campinas, SP, Brazil.
authors.forename: Diogo N
authors.initials: DN
authors.lastname: de Oliveira
authors.affiliation: Brazilian Biosciences National Laboratory, National Energy and Material Research Center, Post Office box: 6192, 13083-877, Campinas, SP, Brazil.
authors.forename: Rafael
authors.initials: R
authors.lastname: Canevarolo
authors.affiliation: Achilles Genetics Ltda, Rua Padre de Toledo Leite, 20 - Centro, 17400-000, Garça, SP, Brazil.
authors.forename: Marcos A
authors.initials: MA
authors.lastname: Achilles
authors.affiliation: Achilles Genetics Ltda, Rua Padre de Toledo Leite, 20 - Centro, 17400-000, Garça, SP, Brazil.
authors.forename: Daniela L
authors.initials: DL
authors.lastname: D'Ercole
authors.affiliation: Laboratory for Veterinary Physiology and Biochemistry, Department of Veterinary Sciences, Faculty of Pharmaceutical, Biomedical and Veterinary Sciences, University of Antwerp, Universiteitsplein 1 Gebouw U 0.09, B-2610, Wilrijk, Belgium.
authors.forename: Peter E
authors.initials: PE
authors.lastname: Bols
authors.affiliation: Department of Animal Reproduction, College of Veterinary Medicine and Animal Science, São Paulo University, Av. Prof. Dr. Orlando Marques de Paiva, 87 - Cidade Universitária, 05508-270, São Paulo, SP, Brazil.
authors.forename: Jose A
authors.initials: JA
authors.lastname: Visintin
authors.affiliation: Department of Animal Science, College of Agricultural Sciences, The Pennsylvania State University, 324 Henning Building University Park, PA, 16802, USA.
authors.forename: Gary J
authors.initials: GJ
authors.lastname: Killian
authors.affiliation: Innovare Biomarkers Laboratory, Medicine and Experimental Surgery Nucleus, University of Campinas, Rua Cinco de Junho, 350 - Barão Geraldo, 13083-877, Campinas, SP, Brazil.
authors.forename: Rodrigo R
authors.initials: RR
authors.lastname: Catharino
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D001755:Blastocyst / Q000378:metabolism*; D002417:Cattle; D046149:Embryo Culture Techniques; D047108:Embryonic Development; D005260:Female; D005307:Fertilization in Vitro; D059471:In Vitro Oocyte Maturation Techniques; D055432:Metabolomics / Q000379:methods*; D015999:Multivariate Analysis; D009865:Oocytes / Q000378:metabolism*; D010713:Phosphatidylcholines / Q000378:metabolism; D011247:Pregnancy; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013109:Sphingomyelins / Q000378:metabolism; D013997:Time Factors
chemical_list: D015415:Biomarkers; D010713:Phosphatidylcholines; D013109:Sphingomyelins