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Published 17 September 2001. doi:10.1083/jcb.200103010
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© The Rockefeller University Press, 0021-9525/2001/9/1235 $5.00
The Journal of Cell Biology, Volume 154, Number 6, September 17, 2001 1235-1244

Article

 Histamine treatment induces rearrangements of orthogonal arrays of particles (OAPs) in human AQP4-expressing gastric cells

Monica Carmosino1, Giuseppe Procino1, Grazia Paola Nicchia1, Roberta Mannucci2, Jean-Marc Verbavatz3, Renèe Gobin3, Maria Svelto1 and Giovanna Valenti1

1 Dipartimento di Fisiologia Generale ed Ambientale, Università degli Studi, 70126 Bari, Italy
2 Sezione di Medicina Interna ed Oncologia, Dipartimento di Medicina Clinica e Sperimentale, Università degli Studi, 06100 Perugia, Italy
3 Service de Biologie Cellulaire, Centre d'Etudes de Saclay, F-91191 Gif sur Yvette Cedex, France

Address correspondence to Giovanna Valenti, Dipartimento de Fisiologia Generale ed Ambientale, Via Amendola 165/A, 70126 Bari, Italy. Tel.: 39-080-544-3444. Fax: 39-080-544-3388. E-mail: g.valenti{at}biologia.uniba.it

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°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 ~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.

Key Words: aquaporin; stomach; water channel; histamine; H+/K+-ATPase


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