title: Effect of different culture systems on adipocyte differentiation-related protein (ADRP) in bovine embryos.
abstract: Bovine embryos cultured in serum-containing media abnormally accumulate lipid droplets, compared to their in vivo counterparts. The objective of this study was to investigate the effect of different culture systems on the mRNA expression and on the quantification and localisation of adipocyte differentiation-related protein (ADRP), a protein associated with lipid accumulation in bovine blastocysts. Two experiments were independently performed for ADRP mRNA expression analysis. In experiment A, blastocysts were produced in modified synthetic oviduct fluid (mSOF)+10% foetal calf serum (FCS), in coculture (bovine oviduct epithelial cells, Boec) and in ewe oviducts, whereas in experiment B, they were produced in mSOF+10μM docosahexaenoic acid (DHA) and in vivo. Control groups were also performed. ADRP mRNA expression was downregulated in the Boec, ewe oviduct and in vivo groups compared to the 10% FCS or DHA groups, respectively. Moreover, the expression of this protein was downregulated in the Boec group compared to the control group (P<0.05). A third experiment (experiment C) was performed to quantify and localise ADRP protein. Boec, in vivo and control groups were tested. After immunofluorescence staining followed by confocal microscopy analysis, embryonic ADRP was clearly localised around lipid droplets, indicating that ADRP is also a lipid droplet coat protein in bovine embryos. In conclusion, our results demonstrate that bovine embryos at the blastocyst stage expressed ADRP mRNA and protein, and that the embryonic culture system modified this expression.
authors.affiliation: INRA, UMR85 Physiologie de la Reproduction et des Comportements, F-37380 Nouzilly, France; CNRS, UMR7247 Physiologie de la Reproduction et des Comportements, F-37380 Nouzilly, France; Université François Rabelais de Tours, F-37041 Tours, France; IFCE, F-37380 Nouzilly, France.
authors.forename: A
authors.initials: A
authors.lastname: Al Darwich
authors.affiliation: INRA, UMR85 Physiologie de la Reproduction et des Comportements, F-37380 Nouzilly, France; CNRS, UMR7247 Physiologie de la Reproduction et des Comportements, F-37380 Nouzilly, France; Université François Rabelais de Tours, F-37041 Tours, France; IFCE, F-37380 Nouzilly, France.
authors.forename: C
authors.initials: C
authors.lastname: Perreau
authors.affiliation: INRA, UMR85 Physiologie de la Reproduction et des Comportements, F-37380 Nouzilly, France; CNRS, UMR7247 Physiologie de la Reproduction et des Comportements, F-37380 Nouzilly, France; Université François Rabelais de Tours, F-37041 Tours, France; IFCE, F-37380 Nouzilly, France.
authors.forename: G
authors.initials: G
authors.lastname: Tsikis
authors.affiliation: INRA, UMR85 Physiologie de la Reproduction et des Comportements, F-37380 Nouzilly, France; CNRS, UMR7247 Physiologie de la Reproduction et des Comportements, F-37380 Nouzilly, France; Université François Rabelais de Tours, F-37041 Tours, France; IFCE, F-37380 Nouzilly, France.
authors.forename: E
authors.initials: E
authors.lastname: Coudert
authors.affiliation: INRA, UMR85 Physiologie de la Reproduction et des Comportements, F-37380 Nouzilly, France; CNRS, UMR7247 Physiologie de la Reproduction et des Comportements, F-37380 Nouzilly, France; Université François Rabelais de Tours, F-37041 Tours, France; IFCE, F-37380 Nouzilly, France.
authors.forename: J L
authors.initials: JL
authors.lastname: Touzé
authors.affiliation: INRA, UEPAO, F-37380 Nouzilly, France.
authors.forename: E
authors.initials: E
authors.lastname: Briant
authors.affiliation: Université de Liège, Faculté de Médecine Vétérinaire, Physiologie de la Reproduction, B4000 Liège, Belgium.
authors.forename: J F
authors.initials: JF
authors.lastname: Beckers
authors.affiliation: INRA, UMR85 Physiologie de la Reproduction et des Comportements, F-37380 Nouzilly, France; CNRS, UMR7247 Physiologie de la Reproduction et des Comportements, F-37380 Nouzilly, France; Université François Rabelais de Tours, F-37041 Tours, France; IFCE, F-37380 Nouzilly, France.
authors.forename: P
authors.initials: P
authors.lastname: Mermillod
authors.affiliation: INRA, UMR85 Physiologie de la Reproduction et des Comportements, F-37380 Nouzilly, France; CNRS, UMR7247 Physiologie de la Reproduction et des Comportements, F-37380 Nouzilly, France; Université François Rabelais de Tours, F-37041 Tours, France; IFCE, F-37380 Nouzilly, France. Electronic address: Florence.guignot@tours.inra.fr.
authors.forename: F
authors.initials: F
authors.lastname: Guignot
mesh_terms: D000818:Animals; D002417:Cattle / Q000196:embryology*; D018920:Coculture Techniques / Q000662:veterinary; D046149:Embryo Culture Techniques / Q000662:veterinary*; D004622:Embryo, Mammalian / Q000378:metabolism*; D005307:Fertilization in Vitro; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D000070780:Perilipin-2; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012756:Sheep
chemical_list: D008565:Membrane Proteins; D000070780:Perilipin-2; D012333:RNA, Messenger
keywords: ADRP quantification and localisation; Culture environment; Lipid droplet