Changes in membrane fluidity and Na+/K(+)-ATPase activity during human trophoblast cell culture.
The human placenta plays an essential role in embryo development, in particular regulating the transport of ions, nutrients and immunoglobulins from the maternal to the fetal circulation. Trophoblast organization into a syncytial layer involves structural and functional steps that may be monitored and elucidated by in vitro studies. The structural stages by which the syncytial trophoblast is formed are not yet understood. In order to clarify the mechanism of trophoblast development, we studied the morphological characteristics of the syncytial trophoblast formation in culture and the functional changes (transport properties and membrane microviscosity) accompanying the structural modifications. By using both 5-nitroxystearate and 16-nitroxystearate as spin labels, we observed an initial increase in membrane order over 0-24 h of culture, which can be associated with two events: recovery of cell membranes from trypsin and initial aggregation of cytotrophoblasts. The similar behaviour of the order parameters determined with both probes indicates that membrane order changes both inside and in the outer part of the lipid bilayer. The subsequent decrease in membrane order observed at 36-48 h might be related to the process of cellular fusion. The increase in sodium/potassium pump activity in the first 24 h of culture might be an expression of cell recovery following trypsin treatment. The subsequent decrease might represent an adaptive mechanism by which metabolic energy is mainly used for morphogenetic changes.
Mazzanti
L
L
Institute of Biochemistry, School of Medicine, University of Ancona, Italy.
Rabini
R A
RA
Biagini
G
G
Pugnaloni
A
A
de Pirro
R
R
Faloia
E
E
Mancini
V
V
Romanini
C
C
Cester
N
N
D002478:Cells, Cultured; D003497:Cyclic N-Oxides; D004578:Electron Spin Resonance Spectroscopy; D006801:Humans; D008560:Membrane Fluidity; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism*; D013113:Spin Labels; D013997:Time Factors; D014327:Trophoblasts / Q000502:physiology* / Q000648:ultrastructure
D003497:Cyclic N-Oxides; D013113:Spin Labels; C005072:5-doxylstearic acid; C032363:16-nitroxystearic acid; D000254:Sodium-Potassium-Exchanging ATPase