Metabolomic analysis revealed differences between bovine cloned embryos with contrasting development abilities. 31. Colloque Scientifique de l'AET

Brochard, Vincent

Dube, Delphine

Duranthon, Véronique

Fernandez-Gonzales, A.

Gomez, E.

Laloë, Denis

Le Bourhis, Daniel

Martin, D.

Munoz, M.

Trigal, B.

English

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Metabolomic analysis revealed differences between
bovine cloned embryos with contrasting development
abilities
Denis Laloë, Daniel Le Bourhis, Vincent Brochard, A. Fernandez-Gonzales,
Delphine Dube, B. Trigal, M. Munoz, D. Martin, E. Gomez, Véronique
Duranthon
To cite this version:
Denis Laloë, Daniel Le Bourhis, Vincent Brochard, A. Fernandez-Gonzales, Delphine Dube, et al..
Metabolomic analysis revealed differences between bovine cloned embryos with contrasting devel-
opment abilities. 31. Colloque Scientifique de l’AETE, Association Européenne de Transfert Em-
bryonnaire - European Embryo Transfer Association (AETE). FRA., Sep 2015, Ghent, Belgium.
￿hal-01536465￿
31st Annual Meeting A.E.T.E. – Ghent, Belgium, 11th – 12th September 2015 140 
METABOLOMIC ANALYSIS REVEALED DIFFERENCES BETWEEN BOVINE 
CLONED EMBRYOS WITH CONTRASTING DEVELOPMENT ABILITIES 
 
LALOË D1, LE BOURHIS D2,3, BROCHARD V3, FERNANDEZ-GONZALEZ A5 , DUBE D3, 
TRIGAL B4, MUNOZ M4, MARTIN D4, GOMEZ E4, DURANTHON V3 
 
1. INRA, UMR 1313 Génétique Animale et Biologie Intégrative, F-78350 Jouy en Josas, France;  
2. Allice, Station de Phénotypage, Lieu-Dit Le Perroi 37380 Nouzilly, France. 
3. INRA, UMR1198 Biologie du Développement et Reproduction, F-78350 Jouy-en-Josas, France; 
4. Centro de Biotecnologíca Animal-SERIDA, Camino de Rioseco 1225, Gijon,  
33394 Asturias, Spain; 
5. Servicios Cientifico Tecnicos (Oviedo), Universidad de Oviedo, Oviedo, 33006 Asturias, Spain. 
 
 
Cloning by somatic cell nuclear transfer has been established in various species but its 
efficiency remains low. Efforts have been made to improve cloning efficiency mainly focusing on 
reprogramming, using donor cells sourced from different somatic tissues and   using epigenetic 
modifiers. Differences in reprogramming efficiency linked to the genotype of donor nuclei have 
been observed in mouse and bovine, but remained largely unexplained, especially in the bovine. 
This study is part of a set of analysis to understand the early differences between two types of 
bovine cloned blastocysts with different term development abilities (2.5 versus 12.5% birth rates) 
(Bui, Reproduction, 138(2), 289-99, 2009). Blastocysts were obtained by nuclear transfer of 
fibroblasts derived from ear skin of two different Holstein heifers (OV 0029 and OV5538) as 
described (Khan, PlosOne, 7(3), e34110, 2012). After activation, embryos were cultured in groups 
in SOF medium with 1% oestrus cow serum (39°C, 5%CO2, 5%O2). On Day 6 embryos were 
cultured individually for 24 hours in 12µl droplets of SOF (Minitube) plus Bovine Serum Albumin.  
On Day 7, 10µl spent culture medium (CM) and blank samples were collected and frozen until 
Fourier Transform Infrared Spectroscopy (FTIR) analysis. The metabolic fingerprint of spent CM 
of the two types of cloned blastocysts were compared. Briefly, samples (n=36 and 65, respectively 
for OV0029 and OV5538) were analyzed using a Golden-Gate ATR device mounted on a Varian 
620-IR FTIR spectrophotometer. After correction for experimental series and subtraction of values 
obtained for blank samples from experimental values, the FTIR spectra were analyzed by 
redundancy analysis (RDA), a method which combines multiple regression with Principal 
Component Analysis. The model included embryo stage at day7, embryo grade at day7 (according 
to IETS grading; only grade 1 and 2 embryos) and embryo genotype. Significance of the effect 
"embryo genotype" was addressed with permutations tests. Analysis on the whole spectra did not 
show significant effects. However, focusing on sub-regions of the spectra pointed to a significant 
"embryo genotype" effect (p=0.035) for wavelengths between 2850 and 3030cm-1. The above 
identified region of the spectra covers the CH3 and CH2 asymmetric stretching from lipids 
(Socrates, Infrared and Raman Characteristic Group Frequencies, Tables and Charts, Ed. Wiley, 
2001 ch 23). Therefore, our results suggest differences in lipid metabolism between these two types 
of clones. Further analysis is in progress to confirm this hypothesis. 
 
 
 
Supported by AGL2012-37772 and FEDER  
  


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