Effect of PUFA on embryo cryoresistance, gene expression and AMPKalpha phosphorylation in IVF-derived bovine embryos.
The objectives of the present study were to evaluate the effect of conjugated linoleic acid (CLA t10, c12, C18:2), linolenic acid (C18:3) and docosahexaenoic acid (DHA, C22:6) supplementation on in vitro bovine embryo development, embryo survival after cryopreservation, gene expression and AMPKalpha phosphorylation. Control groups with modified synthetic oviduct fluid (mSOF)+/-100microM beta-mercaptoethanol (beta-ME) were performed. The effects of co-culture with bovine oviduct epithelial cell (Boec) monolayers, serum supplementation and embryo development in the ewe oviduct, on gene expression were also examined. Experiments 1 and 2: a lower d 7 embryo survival was found with 100microM C22:6 and 100microM C18:2 supplementation compared to 1microM C22:6 and 100microM beta-ME supplementation (P<0.05). C18:3 supplementation had no effect on d 7 embryo survival, but 100microM C18:3 increased d 8 embryo survival compared to 100microM beta-ME supplementation (P<0.05). Experiments 3 and 4: stearoyl-CoA desaturase 1 (SCD1) and sterol regulatory element-binding transcription factor 1 (SREBP1) mRNA decreased after 10microM C22:6 supplementation compared to all other supplementations (P<0.05). A lower fatty acid desaturase 2 (FADS2) transcript level was found with 100microM C18:2, 10microM C22:6 and 10microM C18:3 supplementations compared to groups without fatty acid supplementation (P<0.05). Acetyl-CoA-carboxylase (ACC), fatty acid synthase (FAS), adipose differentiation-related protein (ADRP), acyl-CoA synthetase long-chain family member 1 (ACSL1), diacylglycerol O-acyltransferase 1 (DGAT1), carnitin palmitoyltransferase-II (CPT-II) mRNAs expression and AMPKalpha phosphorylation were not modified with PUFA supplementation. Experiment 5: SCD1 and FAS mRNA decrease in Boec group compared to serum supplementation, as SCD1 mRNA in ewe oviduct group (P<0.05). In conclusion, this study showed that a PUFA supplementation with C18:2, C18:3 or C22:6 in bovine culture development for 6 days and co-culture with Boec down-regulate mRNA expression of proteins involved in lipid metabolism in d 7-8 embryo (SCD1 and FADS2 desaturases), probably through SREBP1 mRNA regulation after 10microM C22:6 supplementation, indicating a modification of saturated/unsaturated fatty acid balance in bovine blastocyst.
Al Darwich
Abdulrahman
A
INRA, UMR85 Physiologie de la Reproduction et des Comportements, F-37380 Nouzilly, France.
Perreau
Christine
C
Petit
Marie Hélène
MH
Papillier
Pascal
P
Dupont
Joëlle
J
Guillaume
Daniel
D
Mermillod
Pascal
P
Guignot
Florence
F
D055372:AMP-Activated Protein Kinases / Q000235:genetics*; D000818:Animals; D002417:Cattle; D015925:Cryopreservation; D004622:Embryo, Mammalian / Q000378:metabolism; D005231:Fatty Acids, Unsaturated / Q000494:pharmacology*; D005260:Female; D005307:Fertilization in Vitro; D015870:Gene Expression / Q000187:drug effects*; D010766:Phosphorylation / Q000187:drug effects; D012333:RNA, Messenger / Q000378:metabolism; D051780:Sterol Regulatory Element Binding Protein 1 / Q000235:genetics / Q000378:metabolism
D005231:Fatty Acids, Unsaturated; D012333:RNA, Messenger; D051780:Sterol Regulatory Element Binding Protein 1; D055372:AMP-Activated Protein Kinases