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doi: 10.1242/10.1242/jcs.00053

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Journal of Cell Science 115, 3667-3674 (2002)
© 2002 The Company of Biologists Limited
DOI: 10.1242/jcs.00053

Research Article

Functional involvement of VAMP/synaptobrevin-2 in cAMP-stimulated aquaporin 2 translocation in renal collecting duct cells

Sabine Gouraud*,1, Antonia Laera*,1, Giuseppe Calamita1, Monica Carmosino1, Giuseppe Procino1, Ornella Rossetto2, Roberta Mannucci3, Walter Rosenthal4, Maria Svelto1 and Giovanna Valenti1,{ddagger}

1 Dipartimento di Fisiologia Generale ed Ambientale, University of Bari, Via Amendola 165/A, 70126 Bari, Italy
2 Centro CNR Biomembrane and Dipartimento di Scienze Biomediche, Via G. Colombo 3, 35100 Padova, Italy
3 Sezione di Medicina Interna ed Oncologia, Dipartimento di Medicina Clinica e Sperimentale, Università degli Studi, Policlinico Monteluce, 06100 Perugia, Italy
4 Forschungsinstitut für Molekulare Pharmakologie, 13125 Berlin, Germany

{ddagger} Author for correspondence (e-mail: g.valenti{at}biologia.uniba.it)

Accepted 10 July 2002

The involvement of soluble N-ethylmaleimide sensitive factor-attachment protein receptor (SNARE) proteins in the cAMP-induced exocytosis of aquaporin 2 (AQP2)-containing vesicles was investigated in AQP2-transfected renal CD8 cells. RT-PCR and western blot analysis confirmed the presence of the SNARE homologs VAMP/synaptobrevin-2, syntaxin-1, syntaxin-4 and SNAP-23 in CD8 cells. Tetanus neurotoxin (TeNT) was efficient in cleaving synaptobrevin-like protein both in vitro and in intact CD8 cells incubated with the toxin. TeNT treatment in intact CD8 cells completely abolished cAMP-stimulated AQP2 targeting to the plasma membrane, as assessed by quantification of cell-surface immunoreactivity to an anti-AQP2 antibody raised against a peptide reproducing the extracellular AQP2 C-loop.

These results represent the first evidence for the functional involvement of VAMP-2 in cAMP-induced AQP2 exocytosis in renal cells.

Key words: Aquaporin, SNARE, Tetanus toxin, Trafficking, Exocytosis


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© The Company of Biologists Ltd 2002