LIPIDOMIC ANALYSIS OF IN VITRO PRODUCED BOVINE EMBRYOS WITH DIFFERENT
DEVELOPMENTAL KINETICS
K. Annes, C.A. Soares, E.C. dos Santos, T. da Silva, D.L Latorre Oliveira, M.J
Sudano, A. Tata,
V.G. Santos, K.R.A. Belaz, M.N. Eberlin, M.P. Milazzotto
Centro de Ciências Naturais e Humanas, Universidade Federal do ABC; Laboratório de
Espectrometria de Massa ThoMSon, Instituto de Química, Universidade de Campinas; Laboratório
de Genética e Melhoramento Animal, Universidade Federal do Pampa; FASB, Faculdade São
Bernardo, Brazil
Lipid accumulation in IVP embryos has been related with a lower cryopreservation efficiency and
developmental potential, been an indicative of inadequate in vitro culture conditions when
compared with in vivo system. The hypothesis of this work is that embryos with different
developmental kinetics metabolize and pile up lipids differentially, which may be reflected in the
embryo viability. So, our objective was to gather quantitative (by SUDAN BLACK B staining) and
qualitative (by MALDI - Matrix Assisted Laser Desorption Ionization – Mass Spectrometry) data
about the lipids in embryos of fast (4 cells 40hpi) and slow (2 cells 40hpi) development. IVP
bovine embryos were produced and cultured individually. They were classified in fast and slow
and analyzed at cleavage, morulae and blastocyst stages. For MALDI analysis, in vivo blastocysts
were included as control group. The data were submitted to ANOVA with Tukey post test (SUDAN
BLACK) or FlexAnalysis (MALDI). There was no difference in the amount of lipids between
cleavage and morulae groups, however, slow blastocyst presented lower amounts when
compared to the fast ones. MALDI results revealed a total of 15 metabolites for all the groups. At
cleavage stage no differences were observed about the quantity of these metabolies. These
differences started to appear at morula stage and were more evident at blastocyst stage.
Fragments at m/z between 758,8 and 788,8 were the most variable among groups, while
fragments at m/z between 808,9 and 810,8 were the most preserved. These results highlight the
developmental kinetics and culture conditions having direct influence on lipid accumulation and
metabolism in IVP bovine embryos. Acknowledgements: FAPESP, CAPES and UFABC.