@article{oai:niigata-u.repo.nii.ac.jp:00028381, author = {Ueno, Sayaka and Niimura, Sueo}, issue = {2}, journal = {新潟大学農学部研究報告, 新潟大学農学部研究報告}, month = {Mar}, note = {The size of perivitelline space was measured in mouse oocytes cultured without cumulus cells (DOs) and compared with that in oocytes cultured with cumulus cells. In addition, the size of perivitelline space of DOs cultured in the medium containing 4-methylumbelliferone (MU), an inhibitor of hyaluronan synthase, was observed to examine whether oocytes are involved in the synthesis and secretion of hyaluronan which plays a role in the enlargement of perivitelline space. Of oocytes cultured with and without cumulus cells for 14hrs, 96.7% (29/30) and 93.5% (29/31) held 1st polar bodies in their perivitelline space, respectively. The percentages of oocytes with 1st polar bodies in the perivitelline space (maturation rates) did not differ between those cultured with and without cumulus cells. The mean size of perivitelline space in oocytes cultured with cumulus cells (5.40μm) did not differ from that (5.08μm) of DOs. When DOs were cultured in the medium containing 0.25mM MU, 88.2% of the oocytes held 1st polar bodies in the perivitelline space, showing no difference from the percentage (85.7%) of control DOs cultured in the medium without MU. The mean size of perivitelline space in DOs treated with 0.25mM MU was 3.58μm, which was significantly smaller than 4.65μm in the control DOs. When DOs were cultured in the media containing 0.25mM MU and hyaluronan at 0.05, 0.25 or 0.50 mg/ml, 86.9 to 92.8% of the DOs held 1st polar bodies in the perivitelline space, showing no difference from the percentage (88.2%) of the DOs cultured in the medium with only 0.25mM MU. The mean sizes of perivitelline space in DOs cultured in the media containing 0.25mM MU and hyaluronan at 0.05 or 0.50mg/ml were 4.24 and 4.29μm, respectively, showing no difference from the size (3.85μm) of the DOs cultured in the medium containing only MU. However, the mean size of perivitelline space in DOs cultured in the medium containing 0.25mM MU and 0.25mg/ml hyaluronan was 4.67μm, which was significantly larger than that of the DOs cultured in the medium containing only MU. These findings strongly suggested that hyaluronan involved in the enlargement of perivitelline space of oocytes is synthesized and secreted by the oocytes themselves., 培養したマウスの裸化卵母細胞(DO)について、囲卵腔の大きさを計測して卵丘卵母細胞複合体(COC)の卵母細胞のものと比較するとともに、ヒアルロナン合成を阻害する作用のある4-メチルウンベリフェロン(MU)で処置したDOの囲卵腔の大きさを観察し、囲卵腔の拡大に役割を果たすヒアルロナンの合成と分泌に卵母細胞が関与しているのかどうかを検討した。採取したDOとCOCを14時間培養したところ、卵母細胞の成熟率は93.5および96.7%であり、両者で有意な差はなかった。DOの囲卵腔は5.08μm あり、COCにおける卵母細胞の5.40μmとの間に有意な差はなかった。また、0.25mMのMUを含むTYH 液で14時間培養したDOの成熟率は88.2%であり、対照のDOの85.7%と有意な差はなかったが、0.25mMのMUを含む培養液で成熟させたDOの囲卵腔は3.58μmであり、対照のDOの4.65μmに比べて有意に小さかった。さらに、0.25mMのMUとともに1ml中に0.05、0.25あるいは0.5mg のヒアルロナンを含む培養液で14時間培養したDOの成熟率は86.9ないし92.8%であり、0.25mMのMUのみで処置したDOの成熟率(88.2%)と相違なかった。一方囲卵腔は、MU のみで処置したDOでは3.85μmあり、MUとともに0.05あるいは0.5mg/mlのヒアルロナンを添加した培養液で培養したDOの4.24および4.29μmとの間に有意差はなかったが、MUとともに0.25mg/mlのヒアルロナンを添加した培養液で培養したDOでは有意に大きく、4.67μmになった。以上の結果から、囲卵腔の拡大に役割を果たすヒアルロナンは卵母細胞自身が合成・分泌していることが推察された。}, pages = {127--133}, title = {Comparison of the Size of Perivitelline Space in Mouse Oocytes Cultured with and without Cumulus Cells and Possible Hyaluronan Production in Oocytes}, volume = {61}, year = {2009} }