Effect of the meiotic inhibitor cilostamide on resumption of meiosis and cytoskeletal distribution in buffalo oocytes.
Improving the quality of in vitro maturated buffalo oocytes is essential for embryo production. We report here the effects on microtubules and microfilaments in oocytes and embryo development that result from treating buffalo oocytes with the phosphodiesterase 3 (PDE3) inhibitor cilostamide. Addition of 20μM or 50μM cilostamide for 24h during in vitro maturation showed no differences in the percentage of oocytes arrested at the germinal vesicle (GV) stage. When 20μM cilostamide was added to the pre-maturation culture for 6h, 12h or 24h and continued for another 24h without cilostamide, oocytes resumed meiosis, but with significantly lower (P<0.01) maturation rates in the 24h group than that in the other two groups. During oocyte maturation in vitro, no microtubules were detected before GV breakdown (GVBD). After GVBD, microtubules combined with condensed chromatin to form the meiotic metaphase spindle. Microfilaments covered a thick area around the cellular cortex and overlying chromosomes. Cilostamide had no effects on microtubules and microfilaments in metaphase II oocytes, and there were no significant differences in the rates of cleavage, blastocyst formation and number of blastocyst cells between oocytes treated pre-maturation with inhibitor for 6h and those of the control group (P>0.05). In summary, cilostamide reversibly arrested the resumption of meiosis without any adverse impact on the dynamic changes in microtubules and microfilaments in buffalo oocytes and their in vitro developmental capacity.
State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresource, Guangxi High Education Laboratory for Animal Reproduction and Biotechnology, Guangxi University, Nanning, Guangxi 530004, China; Department of Reproductive Medicine Center, Dongguan Third People's Hospital, Dongguan, Guangdong 523326, China.
Qing-Yang
QY
Li
State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresource, Guangxi High Education Laboratory for Animal Reproduction and Biotechnology, Guangxi University, Nanning, Guangxi 530004, China.
Juan
J
Lou
State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresource, Guangxi High Education Laboratory for Animal Reproduction and Biotechnology, Guangxi University, Nanning, Guangxi 530004, China.
Xiao-Gan
XG
Yang
State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresource, Guangxi High Education Laboratory for Animal Reproduction and Biotechnology, Guangxi University, Nanning, Guangxi 530004, China.
Yang-Qing
YQ
Lu
State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresource, Guangxi High Education Laboratory for Animal Reproduction and Biotechnology, Guangxi University, Nanning, Guangxi 530004, China. Electronic address: shengshenglu@qq.com.
Sheng-Sheng
SS
Lu
State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresource, Guangxi High Education Laboratory for Animal Reproduction and Biotechnology, Guangxi University, Nanning, Guangxi 530004, China. Electronic address: khlu@gxu.edu.cn.
Ke-Huan
KH
Lu
D008841:Actin Cytoskeleton / Q000187:drug effects; D000818:Animals; D002020:Buffaloes / Q000502:physiology*; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D002478:Cells, Cultured; D003599:Cytoskeleton / Q000187:drug effects* / Q000502:physiology; D005260:Female; D008540:Meiosis / Q000187:drug effects*; D008870:Microtubules / Q000187:drug effects; D009865:Oocytes / Q000187:drug effects* / Q000502:physiology; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology*; D015363:Quinolones / Q000494:pharmacology*; D013997:Time Factors
D010726:Phosphodiesterase Inhibitors; D015363:Quinolones; C021294:cilostamide
Buffalo oocyte; Meiosis; Microfilaments; Microtubules; PDE3 inhibitor