Histamine treatment induces rearrangements of orthogonal arrays of particles (OAPs) in human AQP4-expressing gastric cells.
To test the involvement of the water channel aquaporin (AQP)-4 in gastric acid physiology, the human gastric cell line (HGT)-1 was stably transfected with rat AQP4. AQP4 was immunolocalized to the basolateral membrane of transfected HGT-1 cells, like in native parietal cells. Expression of AQP4 in transfected cells increased the osmotic water permeability coefficient (Pf) from 2.02 +/- 0.3 x 10-4 to 16.37 +/- 0.5 x 10-4 cm/s at 20 degrees C. Freeze-fracture EM showed distinct orthogonal arrays of particles (OAPs), the morphological signature of AQP4, on the plasma membrane of AQP4-expressing cells. Quantitative morphometry showed that the density of OAPs was 2.5 +/- 0.3% under basal condition and decreased by 50% to 1.2 +/- 0.3% after 20 min of histamine stimulation, mainly due to a significant decrease of the OAPs number. Concomitantly, Pf decreased by approximately 35% in 20-min histamine-stimulated cells. Both Pf and OAPs density were not modified after 10 min of histamine exposure, time at which the maximal hormonal response is observed. Cell surface biotinylation experiments confirmed that AQP4 is internalized after 20 min of histamine exposure, which may account for the downregulation of water transport. This is the first evidence for short term rearrangement of OAPs in an established AQP4-expressing cell line.
Carmosino
M
M
Dipartimento di Fisiologia Generale ed Ambientale, Università degli Studi, 70126 Bari, Italy.
Procino
G
G
Nicchia
G P
GP
Mannucci
R
R
Verbavatz
J M
JM
Gobin
R
R
Svelto
M
M
Valenti
G
G
D000818:Animals; D051401:Aquaporin 4; D020346:Aquaporins / Q000378:metabolism*; D002460:Cell Line; D005576:Colforsin / Q000494:pharmacology; D019281:Dimerization; D004847:Epithelial Cells / Q000737:chemistry / Q000648:ultrastructure; D005614:Freeze Fracturing; D006632:Histamine / Q000494:pharmacology*; D006801:Humans; D008854:Microscopy, Electron; D009995:Osmosis / Q000187:drug effects; D010316:Particle Size; D051381:Rats; D013270:Stomach / Q000166:cytology*; D014162:Transfection
C496030:AQP4 protein, human; C496032:Aqp4 protein, rat; D051401:Aquaporin 4; D020346:Aquaporins; D005576:Colforsin; D006632:Histamine