Analysis and characterisation of bovine oocyte and embryo biomarkers by matrix-assisted desorption ionisation mass spectrometry imaging

Autor(es)

Goncalves, Roseli F. ^[1] ; Ferreira, Monica S. ^[2] ; de Oliveira, Diogo N. ^[2] ; Canevarolo, Rafael ^[3] ; Achilles, Marcos A. ^[4] ; D'Ercole, Daniela L. ^[4] ; Bols, Peter E. ^[5] ; Visintin, Jose A. ^[1] ; Killian, Gary J. ^[6] ; Catharino, Rodrigo R. ^[2]

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Dept Anim Reprod, Coll Vet Med & Anim Sci, BR-05508270 Sao Paulo, SP - Brazil

^[2] Univ Estadual Campinas, Innovare Biomarkers Lab, Med & Expt Surg Nucleus, BR-13083877 Campinas, SP - Brazil

^[3] Natl Energy & Mat Res Ctr, Brazilian Biosci Natl Lab, BR-13083877 Campinas, SP - Brazil

^[4] Achilles Genet Ltda, BR-17400000 Garca, SP - Brazil

^[5] Univ Antwerp, Lab Vet Physiol & Biochem, Fac Pharmaceut Biomed & Vet Sci, Dept Vet Sci, B-2610 Antwerp - Belgium

^[6] Penn State Univ, Coll Agr Sci, Dept Anim Sci, University Pk, PA 16802 - USA

• Tipo de documento: Artigo Científico
• Fonte: REPRODUCTION FERTILITY AND DEVELOPMENT; v. 28, n. 3, p. 293-301, 2016.
• Citações Web of Science: 5

Resumo

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. (AU)

• Processo FAPESP: 11/18085-7 - Análise do perfil protéico e lipídico de embriões pré-implantacionais bovinos obtidos por fecundação in vitro, transferência de embriões e por transferência nuclear de células somáticas pela técnica de espectrometria de massas
• Beneficiário: Roseli Fernandes Gonçalves
• Linha de fomento: Bolsas no Brasil - Apoio a Jovens Pesquisadores
• Processo FAPESP: 10/01077-9 - Análise do perfil protéico e lipídico de embriões pré-implantacionais bovinos obtidos por fecundação in vitro, transferência de embriões e por transferência nuclear de células somáticas pela técnica de espectrometria de massas
• Beneficiário: Roseli Fernandes Gonçalves
• Linha de fomento: Auxílio à Pesquisa - Apoio a Jovens Pesquisadores