논문 상세보기
pp.191-200
체외 배양 과정 중에 나타나는 생쥐 초기 2-세포 배의 "in vitro 2-cell block" 현상은 세포내 농도 변화와 밀접한 관련이 있다. 다양한 종류의 세포에서 acetylcholine은 세포막에 존재하는 muscarnic acetylcholine receptor를 통해 세포내 농도 증가를 유도한다. 본 실험에서는 생쥐 "in vitro 2-cell block" 현상에 있어서 ACh의 영향을 알아보기 위해 세포 내 농도 조절 물질을 처리한 후,
Many studies have shown that the development of mouse early 2-cell embryos in vitro is related with the intracellular changes. In ICR strain mouse, the development of embryos arrests at early 2-cell stage, but the arrested early 2-cell embryos can be rescued by the addition of -related materials. Acetylcholine (ACh) increases intracellular concentration ([]i) via the mAChR-PLC-IP3 pathway in mouse oocytes. We examined whether ACh rescues 2-cell block in mouse. In early 2-cell embryos, ACh increased []i in a dose-dependent manner (p<0.001), and had an effect on rescue of 2-cell block and embryonic development. To identify the signal pathway involved in ACh-induced rescue of 2-cell block, we first applied an agonist of ACh receptor (AChR). Like ACh, carbachol increased intracellular concentration ([]i) and atropine, an antagonist of ACh receptor, blocked the ACh-induced increase. In -free medium, ACh also increased []i, indicating that increased by ACh is mainly released from the intracellular store. The ACh-induced increase was blocked by PLC inhibitor (U73122), ryanodine receptor (RyR) antagonist (dantrolene), and CaM KII inhibitor (KN-93), but not by IP3R antagonists (xestospongin C). These results show that ACh increases intracellular concentration via mAChR/PLC/RyR, and this contributes to the rescue of 2-cell block.
