title: Acetylcholine rescues two-cell block through activation of IP3 receptors and Ca2+/calmodulin-dependent kinase II in an ICR mouse strain.
abstract: Acetylcholine (ACh) causes early activation events in mouse oocytes, but little is known about its precise role in the early embryonic development of mice. We aimed to determine whether and how ACh is capable of rescuing two-cell block in an in vitro culture system. ACh evoked different transient Ca(2+) patterns showing a higher Ca(2+) peak in the two-cell stage embryos (two-cells) than observed in mature oocytes. In early two-cells subjected to an in vitro two-cell block, xestospongin C (Xes-C), an IP3 receptor antagonist, significantly decreased the level of the ACh-induced Ca(2+) increase. The reduction in the ACh-induced Ca(2+) increase by Xes-C in late two-cells was lower than that in early two-cells. Furthermore, KN62 and KN93, both CaMKII inhibitors, were found to reduce the magnitude of the ACh-induced Ca(2+) increase in early two-cells. The addition of ACh to the culture medium showed an ability to rescue in vitro two-cell block. However, the addition of ACh together with both Xes-C and CaMKII inhibitors or with either inhibitor separately had no effect on the rescue of two-cell block. Long-term exposure of late two-cells to ACh decreased morula and early blastocyst development and ACh had a differential effect on early and late two-cells. These results indicate that ACh likely rescues the in vitro two-cell block through activation of IP3R- and/or CaMKII-dependent signal transduction pathways.
authors.affiliation: CHO-A Biotechnology Research Institute, CHO-A Pharmaceutical Company, LTD, Seoul, 150-992, South Korea.
authors.forename: Seong-Keun
authors.initials: SK
authors.lastname: Cho
authors.forename: Sook-Young
authors.initials: SY
authors.lastname: Yoon
authors.forename: Chang-Gi
authors.initials: CG
authors.lastname: Hur
authors.forename: Hae-Young
authors.initials: HY
authors.lastname: Yang
authors.forename: Changyong
authors.initials: C
authors.lastname: Choe
authors.forename: Eun-Jin
authors.initials: EJ
authors.lastname: Kim
authors.forename: Jung Soo
authors.initials: JS
authors.lastname: Joo
authors.forename: Kee Ryeon
authors.initials: KR
authors.lastname: Kang
authors.forename: Jae-Yong
authors.initials: JY
authors.lastname: Park
authors.forename: Seong-Geun
authors.initials: SG
authors.lastname: Hong
authors.forename: Jaehee
authors.initials: J
authors.lastname: Han
authors.forename: Dawon
authors.initials: D
authors.lastname: Kang
mesh_terms: D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / Q000031:analogs & derivatives / Q000494:pharmacology; D000109:Acetylcholine / Q000494:pharmacology*; D000818:Animals; D001596:Benzylamines / Q000494:pharmacology; D020013:Calcium Signaling / Q000187:drug effects; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2 / Q000037:antagonists & inhibitors / Q000502:physiology*; D047108:Embryonic Development / Q000187:drug effects; D005260:Female; D053496:Inositol 1,4,5-Trisphosphate Receptors / Q000187:drug effects / Q000502:physiology*; D047028:Macrocyclic Compounds / Q000494:pharmacology; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D010080:Oxazoles / Q000494:pharmacology; D013449:Sulfonamides / Q000494:pharmacology; D000096926:Benzenesulfonamides
chemical_list: D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D000109:Acetylcholine; D001596:Benzylamines; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D053496:Inositol 1,4,5-Trisphosphate Receptors; D047028:Macrocyclic Compounds; D010080:Oxazoles; D013449:Sulfonamides; C511704:xestospongin C; C072105:KN 93; D000096926:Benzenesulfonamides; C063302:KN 62