Obox4 critically regulates cAMP-dependent meiotic arrest and MI-MII transition in oocytes.
Extra follicular oocytes spontaneously resume meiosis in vitro, but the intact germinal vesicle (GV) is retained if the oocytes are cultured in medium containing phosphodiesterase (PDE) inhibitors or cAMP analogues. On the basis of our finding that Obox4 is prominently expressed in oocytes, the present study was conducted to determine the functional role of the homeodomain-containing factor Obox4 during in vitro oocyte maturation. After microinjection of Obox4 dsRNA into the cytoplasm of GV oocytes cultured in M16 medium, oocytes were arrested at metaphase I (MI, 77.7%) and metaphase II (MII, 22.3%). Surprisingly, however, 89% of Obox4 RNAi-treated oocytes resumed meiosis and developed to MI and MII when cultured in medium containing 0.2 mM 3-isobutyl-1-methyl-xanthine (IBMX), in which untreated oocytes maintain intact GVs. Spindles were aberrant, and chromosomes were severely aggregated with decreased MPF and MAP kinase activities in arrested MI oocytes after exposure to Obox4 RNAi. Oocytes overexpressing Obox4 retained intact GVs when cultured in M16 medium. Taken together, for the first time to our knowledge, these findings indicate that Obox4 plays a key role in the cAMP-dependent signaling cascades that maintain GV arrest. Oocytes not expressing Obox4 failed to maintain intact GVs in IBMX-supplemented medium, while GVs remained intact when oocytes were kept in plain medium and overexpressing Obox4, suggesting that Obox4 plays a critical role in cAMP-dependent cascade for maintaining intact GVs.
Lee
Hyun-Seo
HS
Department of Biomedical Science, College of Life Science, CHA University, 606-13 Yeoksam-1- dong, Gangnam-gu, Seoul 135-081, Korea.
Kim
Eun-Young
EY
Kim
Kyeoung-Hwa
KH
Moon
Jisook
J
Park
Kyung-Soon
KS
Kim
Kwang-Soo
KS
Lee
Kyung-Ah
KA
D000818:Animals; D002875:Chromosomes; D000242:Cyclic AMP; D005260:Female; D018398:Homeodomain Proteins / Q000502:physiology*; D008540:Meiosis; D000090204:Mesothelin; D008677:Metaphase; D051379:Mice; D009865:Oocytes / Q000166:cytology*; D015398:Signal Transduction; D008941:Spindle Apparatus
D018398:Homeodomain Proteins; C000715528:Msln protein, mouse; C499222:Obox4 protein, mouse; D000242:Cyclic AMP; D000090204:Mesothelin