title: Effect of addition of FSH, LH and proteasome inhibitor MG132 to in vitro maturation medium on the developmental competence of yak (Bos grunniens) oocytes.
abstract: BACKGROUND
  The competence for embryonic development after IVF is low in the yak, therefore, we investigated the effects of supplementation of FSH, LH and the proteasome inhibitor MG132 in IVM media on yak oocyte competence for development after IVF.
  
  
  METHODS
  In Experiment 1, yak cumulus-oocyte complexes (COCs) were in vitro matured (IVM) in TCM-199 with 20% fetal calf serum (FCS), 1 microg/mL estradiol-17beta, and different combinations of LH (50 or 100 IU/mL) and FSH (0, 1, 5, 10 microg/mL) at 38.6 degrees C, 5% CO2 in air for 24 h. Matured oocytes were exposed to frozen-thawed, heparin-capacitated yak sperm. Presumptive zygotes were cultured in SOF medium containing 6 mg/ml BSA, 0.5 mg/mL myoinositol, 3% (v/v) essential amino acids, 1% nonessential amino acids and 100 μg/mL L-glutamine (48 h, 38.5 degrees C, 5% CO2, 5% O2, and 90% N2). In Experiment 2, cumulus cells were collected at the end of IVM to determine FSHR and LHR mRNA expression by real-time PCR. In Experiment 3 and 4, COCs were cultured in the presence or absence of the proteasomal inhibitor MG132 from either 0-6 h or 18-24 h after initiation of maturation.
  
  
  RESULTS
  The optimum concentration of FSH and LH in IVM media was 5 microg/mL FSH and 50 IU/mL LH which resulted in the greatest cleavage (79.1%) and blastocyst rates (16.1%). Both FSHR and LHR mRNA were detected in yak cumulus cells after IVM. Treatment with MG132 early in maturation reduced (P<0.05) cleavage and blastocyst rates. Conversely, treatment with MG132 late in maturation improved (P<0.05) blastocyst rate. Optimal results with MG132 were achieved at a concentration of 10 microM.
  
  
  CONCLUSIONS
  An optimum concentration of FSH and LH in IVM medium, and treatment with MG132 late in maturation can improve yak oocytes competence for development after IVF.
authors.forename: Xiao
authors.initials: X
authors.lastname: Xiao
authors.affiliation: College of Life Science and Technology, Southwest University for Nationalities, Chengdu 610041, People's Republic of China. zixd2000@yahoo.com.
authors.forename: Xiang-Dong
authors.initials: XD
authors.lastname: Zi
authors.forename: Hui-Ran
authors.initials: HR
authors.lastname: Niu
authors.forename: Xian-Rong
authors.initials: XR
authors.lastname: Xiong
authors.forename: Jin-Cheng
authors.initials: JC
authors.lastname: Zhong
authors.forename: Jian
authors.initials: J
authors.lastname: Li
authors.forename: Li
authors.initials: L
authors.lastname: Wang
authors.forename: Yong
authors.initials: Y
authors.lastname: Wang
mesh_terms: D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D003470:Culture Media; D005260:Female; D005640:Follicle Stimulating Hormone / Q000008:administration & dosage*; D007976:Leupeptins / Q000008:administration & dosage*; D007986:Luteinizing Hormone / Q000008:administration & dosage*; D009865:Oocytes / Q000187:drug effects* / Q000254:growth & development*; D061988:Proteasome Inhibitors / Q000008:administration & dosage*
chemical_list: D003470:Culture Media; D007976:Leupeptins; D061988:Proteasome Inhibitors; D007986:Luteinizing Hormone; D005640:Follicle Stimulating Hormone; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde
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