@article{oai:niigata-u.repo.nii.ac.jp:00028480,
 author = {Takano, Hiroko and Niimura, Sueo},
 issue = {1},
 journal = {新潟大学農学部研究報告, 新潟大学農学部研究報告},
 month = {Aug},
 note = {Using the enzyme-linked immunosorbent assay and the method of reverse transcription-polymerase chain reaction, cAMP and adenylate cyclase mRNA were demonstrated in porcine oocytes with cumulus cells (COCs) and denuded oocytes (DOs) treated with olomoucine, an inhibitor of cyclin dependent kinase of maturation promoting factor, and the cAMP content and the ability to synthesize cAMP in the treated oocytes and cumulus cells were examined. The fluctuation of cAMP content and the ability to synthesize cAMP in porcine oocytes and cumulus cells cultured for various periods were also examined. The concentration of cAMP in olomoucine-treated COCs was 173 fM, which was significantly lower than 3,067 fM in non-treated COCs, whereas the concentration of cAMP in DOs treated with olomoucine (39 fM) was similar to that in non-treated DOs (24 fM). The concentration of cAMP in cultured DOs was always low and did not significantly change during maturation culture. The concentration of cAMP in cultured COCs was low until 8 hrs of culture, whereas that increased rapidly after 8 hrs of culture and reached the maximum at 12 hrs of culture. The concentration of cAMP in COCs decreased after 22 hrs of culture. The expression of adenylate cyclase mRNA was weaker in COCs and DOs treated with olomoucine than in control COCs and DOs. In cultured COCs and DOs, the expression of such the mRNA was always demonstrated and stronger in those cultured for 8 hrs than in those cultured for 0, 22, 32 and 44 hrs. From these results, it was suggested that cAMP contained in COCs is mostly derived from cumulus cells, although cAMP is synthesized in oocytes. The present findings also suggested that the olomoucine treatment inhibits cAMP synthesis in cumulus cells, which in turn results in the low cAMP level in the cytoplasm of oocytes.卵母細胞成熟促進因子の調節サブユニットであるサイクリンBキナーゼの活性化をオロモウシンで阻害したブタの卵丘卵母細胞複合体(COC)と裸化卵母細胞(DO)について、cAMPおよびcAMPの合成酵素であるアデニル酸シクラーゼのmRNAの検出をELISA法あるいはRT-PCR法を用いて行い、cAMPの含量と合成能に及ぼすオロモウシンの影響を検討した。併せて、各種時間培養したブタのCOCとDOについても、cAMPおよびアデニル酸シクラーゼmRNAの検出を行い、卵丘細胞と卵母細胞におけるcAMPの含量および合成能の変化を調べた。COCが含有するcAMPの濃度は、オロモウシンとともに22時間培養したものでは173fMであり、無処置のものの3,067fMに比べて有意に低かったが､ DOが含有するcAMPの濃度は、オロモウシン処置したもの(39 fM)と無処置のもの(24 fM)との間で相違なかった。またcAMPレベルは、DOでは常に低く、44時間までの培養の経過に伴う変化を示さなかったが、COCでは培養後12ないし22時間に著しく高くなった。一方、アデニル酸シクラーゼmRNAの発現はCOCとDOの両者で認められたが、その発現量は、COCとDOのいずれにおいても、培養後8時間で他の培養時間のものよりも多かった。また、オロモウシン処置したCOCとDOでは、アデニル酸シクラーゼmRNAの発現量は無処置のものに比べて少なかった。以上の結果から、卵母細胞と卵丘細胞の両者はcAMPの合成能を持っているが、COCが含有するcAMPのほとんどは卵丘細胞に由来することが考えられた。またオロモウシン処置は、卵丘細胞でのcAMP合成を抑制することによって、卵母細胞内のcAMPレベルを低下させることがうかがわれた。},
 pages = {15--20},
 title = {The Amount of cAMP in Porcine Oocytes Cultured with Olomoucine},
 volume = {57},
 year = {2004}
}