Expression of calcitonin receptors in mouse preimplantation embryos and their function in the regulation of blastocyst differentiation by calcitonin.
Calcitonin secretion in the pregnant uterus is tightly regulated by the ovarian hormones, estrogen and progesterone, which limit its expression to a brief period preceding blastocyst implantation. The binding of calcitonin to a G protein-coupled receptor activates adenylate cyclase and elevates cytosolic Ca2+ levels. The acceleration of preimplantation embryonic development that is known to occur upon elevation of intracellular Ca2+ prompted an investigation into calcitonin regulation of blastocyst differentiation. Using reverse transcription and the polymerase chain reaction to estimate the relative abundance of calcitonin receptor mRNA, a 25-fold accumulation of the splice variant, CR-1a, was observed in embryos between the 1-cell and 8-cell stages. Cytosolic free Ca2+ levels were rapidly elevated in embryos at the 4-cell to blastocyst stages after exposure to 10 nM calcitonin. Blastocysts treated for 30 minutes with 10 nM calcitonin differentiated in vitro at an accelerated rate, as assessed by the translocation of alpha5beta1 integrin to the apical surface of trophoblast cells, the corresponding elevation of fibronectin-binding activity and the timing of trophoblast cell migration. Chelation of cytosolic free Ca2+ with BAPTA-AM, but not inhibition of protein kinase A activity by H-89, attenuated the effects of calcitonin on blastocyst development. These findings support the concept that calcitonin secretion within the progesterone-primed uterus and the coordinate expression of CR-1a by preimplantation embryos regulates blastocyst differentiation through receptor-mediated Ca2+ signaling.
Wang
J
J
Department of Obstetrics and Gynecology and Department of Anatomy and Cell Biology, C. S. Mott Center for Human Growth and Development, Wayne State University School of Medicine, Detroit, MI 48201-1415, USA.
Rout
U K
UK
Bagchi
I C
IC
Armant
D R
DR
D000818:Animals; D001755:Blastocyst / Q000187:drug effects / Q000378:metabolism*; D002116:Calcitonin / Q000378:metabolism / Q000494:pharmacology / Q000502:physiology*; D002118:Calcium / Q000378:metabolism; D002448:Cell Adhesion; D002614:Chelating Agents / Q000494:pharmacology; D004533:Egtazic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D047108:Embryonic Development; D005314:Embryonic and Fetal Development; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D007546:Isoquinolines / Q000494:pharmacology; D051379:Mice; D018613:Microscopy, Confocal; D011247:Pregnancy; D047428:Protein Kinase Inhibitors; D012333:RNA, Messenger / Q000378:metabolism; D018003:Receptors, Calcitonin / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects; D013449:Sulfonamides; D014599:Uterus / Q000254:growth & development / Q000378:metabolism
D002614:Chelating Agents; D004791:Enzyme Inhibitors; D007546:Isoquinolines; D047428:Protein Kinase Inhibitors; D012333:RNA, Messenger; D018003:Receptors, Calcitonin; D013449:Sulfonamides; C070379:1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester; D004533:Egtazic Acid; D002116:Calcitonin; C063509:N-(2-(4-bromocinnamylamino)ethyl)-5-isoquinolinesulfonamide; D002118:Calcium