Activity and expression of Na+/H+ exchanger isoforms in the syncytiotrophoblast of the human placenta.
The purpose of this study was to compare Na+/H+ exchanger (NHE) activity in the microvillous (MVM) and basal (BM) plasma membrane of the human placental syncytiotrophoblast and to determine the relative contribution of various NHE isoforms to this activity. Uptake of 22Na into isolated MVM vesicles in the presence of a H+ gradient, at initial rate, was four- to fivefold higher than that by BM vesicles (214+/-28 vs. 49+/-9 pmol/mg protein per 30 s, respectively, means+/-SEM, n=8, 6, P<0.001). The 22Na uptake by MVM, but not by BM, was reduced in the absence of a H+ gradient and in the presence of 500 microM amiloride. To determine the contribution of NHE1, NHE2 and NHE3 isoforms to NHE activity in MVM, we investigated the effect of amiloride analogues which show isoform selectivity. HOE 694, an analogue selective for NHE1 at low concentrations, inhibited 22Na uptake with an EC50 of 0.13+/-0.05 microM (n=6), whereas S3226, an analogue selective for NHE3 at low concentrations had an EC50 of 3.01+/-0.85 microM (n=5). To investigate this further, we measured recovery of syncytiotrophoblast intracellular pH (pHi) from an acid load using a H+-selective, fluorescent dye (BCECF) loaded into isolated intact placental fragments. This recovery was blocked in the absence of Na+ and the presence of amiloride (500 microM) and concentrations of HOE 694 and S3226 were comparable to those used in vesicle experiments. Overall these data show that under the conditions used NHE activity in the term placental syncytiotrophoblast is absent from BM. NHE activity in the MVM is attributable predominantly to NHE1.
Speake
P F
PF
Human Development and Reproductive Health Academic Group, Academic Unit of Child Health, University of Manchester, St. Mary's Hospital, Hathersage Road, Manchester, M13 OJH, UK. paul.speake@man.ac.uk
Mynett
K J
KJ
Glazier
J D
JD
Greenwood
S L
SL
Sibley
C P
CP
D027682:Cation Transport Proteins / Q000378:metabolism*; D022162:Cytoplasmic Vesicles / Q000378:metabolism; D005260:Female; D005452:Fluoresceins; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008565:Membrane Proteins / Q000378:metabolism*; D011522:Protons; D012979:Sodium Radioisotopes; D000074082:Sodium-Hydrogen Exchanger 1; D000074083:Sodium-Hydrogen Exchanger 3; D017923:Sodium-Hydrogen Exchangers / Q000378:metabolism*; D014327:Trophoblasts / Q000378:metabolism*
D027682:Cation Transport Proteins; D005452:Fluoresceins; D008565:Membrane Proteins; D011522:Protons; C490998:SLC9A1 protein, human; C081517:SLC9A2 protein, human; C000613385:SLC9A3 protein, human; D012979:Sodium Radioisotopes; D000074082:Sodium-Hydrogen Exchanger 1; D000074083:Sodium-Hydrogen Exchanger 3; D017923:Sodium-Hydrogen Exchangers; C057433:2',7'-bis-(2-carboxyethyl)-5(6)-carboxyfluorescein acetoxymethyl ester