Effect of dynein inhibitor on mouse oocyte in vitro maturation and its cyclin B1 mRNA level.
OBJECTIVE
To evaluate the effect of dynein inhibitor on mouse oocyte in vitro maturation and its cyclin B1 transcription level.


METHODS
Immature mouse oocytes were cultured in vitro with a known dynein ATPase activity inhibitor-sodium orthovanadate (SOV) to detect the changes of maturation rate, and semi-quantitative RT-PCR and single cell RT-PCR were performed to detect the changes of cyclin B1 mRNA level.


RESULTS
In dose-dependent experiments, the maturation rates of oocytes were significantly different between 5 micromol/L SOV and control groups (P < 0.05), and decreased with SOV increasing doses. However, the elevation of cyclin B1 mRNA level of immatured oocytes cultured for 12 h depended on SOV concentrations ranging from 50 to 500 micromol/L. In incontinuity exposed SOV experiments, the maturation rates of oocytes markedly reduced after the first incubation with 400 micromol/L SOV at least for 1 h and were first cultured in SOV-free medium for 4 h or 8 h before exposure to SOV (P < 0.05). In time-course experiment, the opposite changes of cyclin B1 mRNA level in oocytes between SOV and control groups were observed.


CONCLUSIONS
Dynein inhibitor might delay oocytes meiosis process, and cause ectopic expression of cyclin B1 in oocytes. Most Oocytes incubated with SOV blocked at germinal vesicles (GV) stage or M I to anaphase transition due to dynein dysfunction and ectopic transcription level of cyclin B1.
Wang
Xiao-Mei
XM
Department of Cell Biology and Genetics, Research Center for Reproductive Medicine, Shantou University Medical College, Shantou 515041, Guangdong, China. xmwang67@yahoo.com.cn
Huang
Tian-Hua
TH
Xie
Qing-Dong
QD
Zhang
Qing-Jian
QJ
Ruan
Ye
Y
D000818:Animals; D002478:Cells, Cultured; D019926:Cyclin B / Q000235:genetics* / Q000378:metabolism; D056744:Cyclin B1; D004398:Dyneins / Q000037:antagonists & inhibitors; D005260:Female; D005786:Gene Expression Regulation; D008540:Meiosis / Q000187:drug effects*; D051379:Mice; D008807:Mice, Inbred BALB C; D009865:Oocytes / Q000187:drug effects* / Q000254:growth & development / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D014638:Vanadates / Q000494:pharmacology*
C534700:Ccnb1 protein, mouse; D019926:Cyclin B; D056744:Cyclin B1; D012333:RNA, Messenger; D014638:Vanadates; D004398:Dyneins