ATP-stimulated Ca(2+)-activated K(+) efflux pathway and differentiation of human placental cytotrophoblast cells |
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Authors: | Clarson L H Roberts V H J Greenwood S L Elliott A C |
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Institution: | Academic Unit of Child Health, University of Manchester, St. Mary's Hospital, Manchester M13 0JH, United Kingdom. lorraine.carson@man.ac.uk |
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Abstract: | The aim of this study was to determine whether extracellular ATP (ATP](o)) stimulated a Ca(2+)-activated K(+) efflux in trophoblast cells that was dependent on extracellular Ca(2+) (Ca(2+)](o)). Cytotrophoblast cells, isolated from human placenta, were examined following 18 h (relatively undifferentiated) and 66 h (multinucleate cells) of culture. Potassium efflux was measured using (86)Rb as a trace marker. Intracellular Ca(2+) (Ca(2+)](i)) was examined by microfluorometry using fura 2. ATP](o) significantly increased (86)Rb efflux to a peak that declined to control (18-h cells) or an elevated plateau (66-h cells) and was inhibited by 100 nM charybdotoxin. Removing Ca(2+)](o) significantly reduced (86)Rb efflux in both groups as did application of 150 microM GdCl(3). ATP](o) significantly increased Ca(2+)](i) in both groups of cells. The response was reduced by removing Ca(2+)](o) and applying 150 microM GdCl(3). For both (86)Rb efflux and microfluorometry experiments, the response to ATP](o) was more dependent on Ca(2+)](o) in 66-h cells compared with 18-h cells (approximately 70% greater). Cytotrophoblast cells exhibit an ATP](o)-stimulated Ca(2+)-activated K(+) efflux. The dependency of this pathway on Ca(2+)](o) is greater in the 66-h multinucleate syncytiotrophoblast-like cells, suggesting that the mechanism for Ca(2+) entry may be altered during differentiation of trophoblast cells. |
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