title: Glutathione synthesis during in vitro maturation of buffalo (Bubalus bubalis) oocytes: effects of cysteamine on embryo development.
abstract: It was demonstrated that cysteamine supplementation during in vitro maturation (IVM) improves embryo development by increasing glutathione (GSH) synthesis in several species. An improved developmental competence of oocytes matured in the presence of cysteamine was also recorded in buffalo species. The purpose of this work was to investigate (1) if glutathione is de novo synthesized during in vitro maturation of buffalo oocytes, (2) if cysteamine improves buffalo embryo development via an increase in GSH synthesis, and (3) if the inhibition of glutathione synthesis by buthionine sulfoximide (BSO), in the presence or absence of cysteamine, affects subsequent embryo development and GSH synthesis.Cumulus-oocytes complexes (COCs), recovered from slaughtered animals, were matured in vitro in TCM199+10% fetal calf serum (FCS), 0.5 microg/ml FSH, 5 microg/ml LH and 1 microg/ml 17-beta-estradiol in the absence or presence of cysteamine (50 microM), with or without 5mM BSO. Glutathione content was measured by high-performance liquid chromatography (HPLC) and fluorimetric analysis in immature oocytes and in oocytes matured in the different experimental conditions. In a second experiment, the mature oocytes were in vitro fertilized and cultured for 7 days in order to assess development to blastocysts (BLs). It was demonstrated that buffalo oocytes synthesize glutathione during in vitro maturation and that cysteamine increases glutathione synthesis. Furthermore, the promoting effects of cysteamine on embryo development and GSH synthesis were neutralized by buthionine sulfoximide. These results indicate that glutathione plays a critical role on buffalo embryo development.
authors.affiliation: DISCIZIA, Faculty of Veterinary Medicine, Federico II University, 80137 Naples, Italy. bgasparr@unina.it
authors.forename: Bianca
authors.initials: B
authors.lastname: Gasparrini
authors.forename: Hichem
authors.initials: H
authors.lastname: Sayoud
authors.forename: Gianluca
authors.initials: G
authors.lastname: Neglia
authors.forename: Daniel Gustavo de
authors.initials: DG
authors.lastname: Matos
authors.forename: Isabelle
authors.initials: I
authors.lastname: Donnay
authors.forename: Luigi
authors.initials: L
authors.lastname: Zicarelli
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000502:physiology; D002020:Buffaloes / Q000196:embryology* / Q000378:metabolism; D019328:Buthionine Sulfoximine / Q000494:pharmacology; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D003543:Cysteamine / Q000494:pharmacology*; D005314:Embryonic and Fetal Development / Q000187:drug effects*; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary; D005978:Glutathione / Q000096:biosynthesis*; D009865:Oocytes / Q000187:drug effects / Q000502:physiology*; D006080:Ovarian Follicle / Q000166:cytology
chemical_list: D019328:Buthionine Sulfoximine; D003543:Cysteamine; D005978:Glutathione