Intermediate conductance Ca2+-activated K+ channels modulate human placental trophoblast syncytialization

Regulation of human placental syncytiotrophoblast renewal by cytotrophoblast migration, aggregation/fusion and differentiation is essential for successful pregnancy. In several tissues, these events are regulated by intermediate conductance Ca2+-activated K+ channels (IKCa), in part through their ab...

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Bibliographic Details
Published in:PloS one 2014-03, Vol.9 (3), p.e90961-e90961
Main Authors: Díaz, Paula, Wood, Amber M, Sibley, Colin P, Greenwood, Susan L
Format: Article
Language:English
Subjects:
Activation
Analysis of Variance
Antibodies
Benzimidazoles - pharmacology
Biological Transport - drug effects
Biology
Calcium (extracellular)
Calcium channels
Calcium conductance
Cell culture
Cell Culture Techniques
Cell Fusion
Cell size
Channel gating
Channels
Charybdotoxin
Chorionic gonadotropin
Chorionic Gonadotropin - secretion
Cytoplasm
Efflux
Female
Fluorescent Antibody Technique
Gonadotropins
Homeostasis
Humans
Immunofluorescence
In vivo methods and tests
Linear Models
Medicine
Nuclei
Nuclei (cytology)
Pituitary (anterior)
Placenta
Placenta - cytology
Potassium channels
Potassium channels (calcium-gated)
Potassium Channels, Calcium-Activated - drug effects
Potassium Channels, Calcium-Activated - metabolism
Potassium conductance
Preeclampsia
Pregnancy
Pyrazoles - pharmacology
Resistance
Rubidium Radioisotopes - metabolism
Syncytia
Tissues
Trophoblasts - cytology
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Summary:Regulation of human placental syncytiotrophoblast renewal by cytotrophoblast migration, aggregation/fusion and differentiation is essential for successful pregnancy. In several tissues, these events are regulated by intermediate conductance Ca2+-activated K+ channels (IKCa), in part through their ability to regulate cell volume. We used cytotrophoblasts in primary culture to test the hypotheses that IKCa participate in the formation of multinucleated syncytiotrophoblast and in syncytiotrophoblast volume homeostasis. Cytotrophoblasts were isolated from normal term placentas and cultured for 66 h. This preparation recreates syncytiotrophoblast formation in vivo, as mononucleate cells (15 h) fuse into multinucleate syncytia (66 h) concomitant with elevated secretion of human chorionic gonadotropin (hCG). Cells were treated with the IKCa inhibitor TRAM-34 (10 µM) or activator DCEBIO (100 µM). Culture medium was collected to measure hCG secretion and cells fixed for immunofluorescence with anti-IKCa and anti-desmoplakin antibodies to assess IKCa expression and multinucleation respectively. K+ channel activity was assessed by measuring 86Rb efflux at 66 h. IKCa immunostaining was evident in nucleus, cytoplasm and surface of mono- and multinucleate cells. DCEBIO increased 86Rb efflux 8.3-fold above control and this was inhibited by TRAM-34 (85%; p
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0090961