Effect of addition of FSH, LH and proteasome inhibitor MG132 to in vitro maturation medium on the developmental competence of yak (Bos grunniens) oocytes

Reproductive Biology and Endocrinology

Yong Wang¹  Li Wang²  Jian Li¹  Jin-Cheng Zhong¹  Xian-Rong Xiong²  Hui-Ran Niu²  Xiang-Dong Zi²  Xiao Xiao² 

[1] College of Tibetan Plateau Research, Southwest University for Nationalities, Chengdu 610041, People’s Republic of China;College of Life Science and Technology, Southwest University for Nationalities, Chengdu 610041, People’s Republic of China

关键词: Early development;    MG132;    FSH;    Yak;    IVF;   

Others  :  804389
DOI  :  10.1186/1477-7827-12-30

 received in 2014-02-20, accepted in 2014-04-14,  发布年份 2014

【 摘 要 】

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.

【 授权许可】

   
2014 Xiao et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

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