title: Vesicle-associated Membrane Protein 3 (VAMP3) Mediates Constitutive Trafficking of the Renal Co-transporter NKCC2 in Thick Ascending Limbs: ROLE IN RENAL FUNCTION AND BLOOD PRESSURE.
abstract: Renal cells of the thick ascending limb (TAL) reabsorb NaCl via the apical Na+/K+/2Cl-co-transporter NKCC2. Trafficking of NKCC2 to the apical surface regulates NKCC2-mediated NaCl absorption and blood pressure. The molecular mechanisms by which NKCC2 reaches the apical surface and their role in renal function and maintenance of blood pressure are poorly characterized. Here we report that NKCC2 interacts with the vesicle fusion protein VAMP3, and they co-localize at the TAL apical surface. We observed that silencing VAMP3 in vivo blocks constitutive NKCC2 exocytic delivery, decreasing the amount of NKCC2 at the TAL apical surface. VAMP3 is not required for cAMP-stimulated NKCC2 exocytic delivery. Additionally, genetic deletion of VAMP3 in mice decreased total expression of NKCC2 in the TAL and lowered blood pressure. Consistent with these results, urinary excretion of water and electrolytes was higher in VAMP3 knock-out mice, which produced more diluted urine. We conclude that VAMP3 interacts with NKCC2 and mediates its constitutive exocytic delivery to the apical surface. Additionally, VAMP3 is required for normal NKCC2 expression, renal function, and blood pressure.
authors.affiliation: From the Hypertension and Vascular Research Division, Department of Internal Medicine, Henry Ford Hospital, Detroit, Michigan 48202, the Department of Physiology, Wayne State University, Detroit, Michigan 48202, and.
authors.forename: Paulo S
authors.initials: PS
authors.lastname: Caceres
authors.affiliation: From the Hypertension and Vascular Research Division, Department of Internal Medicine, Henry Ford Hospital, Detroit, Michigan 48202.
authors.forename: Mariela
authors.initials: M
authors.lastname: Mendez
authors.affiliation: From the Hypertension and Vascular Research Division, Department of Internal Medicine, Henry Ford Hospital, Detroit, Michigan 48202, the Interim Translational Research Institute, Academic Health System, Hamad Medical Corporation, 16060 Doha, Qatar.
authors.forename: Mohammed Z
authors.initials: MZ
authors.lastname: Haque
authors.affiliation: From the Hypertension and Vascular Research Division, Department of Internal Medicine, Henry Ford Hospital, Detroit, Michigan 48202, the Department of Physiology, Wayne State University, Detroit, Michigan 48202, and portiz1@hfhs.org.
authors.forename: Pablo A
authors.initials: PA
authors.lastname: Ortiz
mesh_terms: D000818:Animals; D001794:Blood Pressure / Q000502:physiology*; D000242:Cyclic AMP / Q000378:metabolism; D005089:Exocytosis / Q000502:physiology; D005786:Gene Expression Regulation / Q000502:physiology; D007668:Kidney / Q000378:metabolism*; D008297:Male; D051379:Mice; D021381:Protein Transport / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley; D012965:Sodium Chloride / Q000378:metabolism; D064466:Solute Carrier Family 12, Member 1 / Q000235:genetics / Q000378:metabolism*; D051040:Vesicle-Associated Membrane Protein 3 / Q000235:genetics / Q000378:metabolism*
chemical_list: C578715:Slc12a1 protein, mouse; C578716:Slc12a1 protein, rat; D064466:Solute Carrier Family 12, Member 1; D051040:Vesicle-Associated Membrane Protein 3; C494953:vesicle-associated membrane protein 3, mouse; C494951:vesicle-associated membrane protein 3, rat; D012965:Sodium Chloride; D000242:Cyclic AMP
keywords: Na-K-Cl cotransporter (NKCC); SNARE proteins; apical surface; blood pressure; epithelial cell; exocytosis; kidney; membrane transport; renal physiology; urine excretion