title: Fatty acid synthesis and oxidation in cumulus cells support oocyte maturation in bovine.
abstract: Oocyte meiotic maturation requires energy from various substrates including glucose, amino acids, and lipids. Mitochondrial fatty acid (FA) β-oxidation (FAO) in the oocyte is required for meiotic maturation, which is accompanied by differential expression of numerous genes involved in FAs metabolism in surrounding cumulus cells (CCs) in vivo. The objective was to elucidate components involved in FAs metabolism in CCs during oocyte maturation. Twenty-seven genes related to lipogenesis, lipolysis, FA transport, and FAO were chosen from comparative transcriptome analysis of bovine CCs before and after maturation in vivo. Using real-time PCR, 22 were significantly upregulated at different times of in vitro maturation (IVM) in relation to oocyte meiosis progression from germinal vesicle breakdown to metaphase-II. Proteins FA synthase, acetyl-coenzyme-A carboxylase, carnitine palmitoyltransferase, perilipin 2, and FA binding protein 3 were detected by Western blot and immunolocalized to CCs and oocyte cytoplasm, with FA binding protein 3 concentrated around oocyte chromatin. By mass spectrometry, CCs lipid profiling was shown to be different before and after IVM. FAO inhibitors etomoxir and mildronate dose-dependently decreased the oocyte maturation rate in vitro. In terms of viability, cumulus enclosed oocytes were more sensitive to etomoxir than denuded oocytes. In CCs, etomoxir (150 μM) led to downregulation of lipogenesis genes and upregulated lipolysis and FAO genes. Moreover, the number of lipid droplets decreased, whereas several lipid species were more abundant compared with nontreated CCs after IVM. In conclusion, FAs metabolism in CCs is important to maintain metabolic homeostasis and may influence meiosis progression and survival of enclosed oocytes.
authors: Institut National de la Recherche Agronomique Unité Mixte de Recherche (UMR) 85, Centre National de la Recherche Scientifique UMR 7247, Université François Rabelais de Tours, and L'Institut français du cheval et de l'équitation, Physiologie de la Reproduction et des Comportements (L.S.-L., D.B., S.E., V.M., V.L., A.D., P.P., P.M., S.U.), F-37380 Nouzilly, France; Electron Microscopy Department (R.U.), AN Belozersky Institute of Physical and Chemical Biology, Moscow State University, 119899 Moscow, Russia; and Laboratoire Biologie Cellulaire et Microscopie Electronique (R.U.), Faculté de Médecine, Université François Rabelais, Tours, 37000 France.
authors: Laura
authors: L
authors: Sanchez-Lazo
authors: Daphné
authors: D
authors: Brisard
authors: Sébastien
authors: S
authors: Elis
authors: Virginie
authors: V
authors: Maillard
authors: Rustem
authors: R
authors: Uzbekov
authors: Valérie
authors: V
authors: Labas
authors: Alice
authors: A
authors: Desmarchais
authors: Pascal
authors: P
authors: Papillier
authors: Philippe
authors: P
authors: Monget
authors: Svetlana
authors: S
authors: Uzbekova
mesh_terms: D021581:Active Transport, Cell Nucleus; D000818:Animals; D002417:Cattle; D002470:Cell Survival; D016000:Cluster Analysis; D054885:Cumulus Cells / Q000378:metabolism*; D005227:Fatty Acids / Q000096:biosynthesis / Q000737:chemistry*; D005260:Female; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D050155:Lipogenesis / Q000235:genetics; D008928:Mitochondria / Q000378:metabolism; D009865:Oocytes / Q000166:cytology*; D009866:Oogenesis / Q000502:physiology; D010053:Ovary / Q000378:metabolism; D010100:Oxygen / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D005227:Fatty Acids; D010100:Oxygen