Store-operated Ca2+ entry in first trimester and term human placenta.
We have examined whether store-operated Ca2+ entry, a common pathway for Ca2+ entry in non-excitable tissue, is apparent in the syncytiotrophoblast of both first trimester and term human placenta. Expression of transient receptor potential (TRPC) homologues, a family of channels thought to be involved in store-operated Ca2+ entry, was also studied at the mRNA and protein levels. [Ca2+]i in syncytiotrophoblast of first trimester and term placental villous fragments was measured by microfluorimetry using the Ca2+-sensitive dye fura-2. Store-operated Ca2+ entry was stimulated using 1 microM thapsigargin in Ca(2+)-free Tyrode buffer (no added Ca2+ + 1 mM EGTA) followed by superfusion with control (Ca2+-containing) buffer. In term fragments, this protocol resulted in a rapid increase in [Ca2+]i, which was inhibited in the presence of 150 microM GdCl3, 200 microM NiCl2, 200 microM CoCl2 or 30 microM SKF96365 but was unaffected by addition of 10 microM nifedipine. It was not possible to stimulate such a rise in [Ca2+]i in first trimester fragments. Messenger RNA encoding TRPC1, TRPC3, TRPC4, TRPC5 and TRPC6 was identified in both first trimester and term placentas. From Western blotting, TRPC3 and TRPC6 proteins were detected in term, but not in first trimester, placentas, while TRPC1 protein was not detected. By immunocytochemistry, TRPC3 and TRPC4 were localised to cytotrophoblast cells in first trimester placentas and to the syncytiotrophoblast in term placentas. TRPC6 staining was present in the syncytiotrophoblast of both first trimester and term placenta, but the intensity was much greater in the latter. We propose that store-operated Ca2+ entry may be an important route for Ca2+ entry into the syncytiotrophoblast of term, but not first trimester placentas, and that in human placenta TRPC channels may underlie this entry mechanism.
Clarson
L H
LH
Academic Unit of Child Health, University of Manchester, St Mary's Hospital, Hathersage Road, Manchester M13 0JH, UK. l.clarson@rowett.ac.uk
Roberts
V H J
VH
Hamark
B
B
Elliott
A C
AC
Powell
T
T
D000328:Adult; D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism*; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D003035:Cobalt / Q000494:pharmacology; D003592:Cytophotometry; D017931:DNA Primers; D004075:Diglycerides / Q000494:pharmacology; D005260:Female; D005470:Fluorometry; D006801:Humans; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D008564:Membrane Potentials / Q000502:physiology; D009532:Nickel / Q000494:pharmacology; D010920:Placenta / Q000166:cytology / Q000378:metabolism*; D011247:Pregnancy; D011261:Pregnancy Trimester, First / Q000378:metabolism*; D011263:Pregnancy Trimester, Third / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013268:Stimulation, Chemical; D014327:Trophoblasts / Q000378:metabolism*
D017931:DNA Primers; D004075:Diglycerides; D012333:RNA, Messenger; D003035:Cobalt; C022838:nickel chloride; D009532:Nickel; C038237:1-oleoyl-2-acetylglycerol; C018021:cobaltous chloride; D002118:Calcium