Renal Na+-K+-Cl- cotransporter activity and vasopressin-induced trafficking are lipid raft-dependent
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Renal Na+-K+-Cl- cotransporter activity and vasopressin-induced trafficking are lipid raft-dependent. / Welker, Pia; Böhlick, Alexandra; Mutig, Kerim; Salanova, Michele; Kahl, Thomas; Schlüter, Hartmut; Blottner, Dieter; Ponce-Coria, Jose; Gamba, Gerardo; Bachmann, Sebastian.
In: AM J PHYSIOL-RENAL, Vol. 295, No. 3, 3, 09.2008, p. 789-802.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - Renal Na+-K+-Cl- cotransporter activity and vasopressin-induced trafficking are lipid raft-dependent
AU - Welker, Pia
AU - Böhlick, Alexandra
AU - Mutig, Kerim
AU - Salanova, Michele
AU - Kahl, Thomas
AU - Schlüter, Hartmut
AU - Blottner, Dieter
AU - Ponce-Coria, Jose
AU - Gamba, Gerardo
AU - Bachmann, Sebastian
PY - 2008/9
Y1 - 2008/9
N2 - Apical bumetanide-sensitive Na(+)-K(+)-2Cl(-) cotransporter (NKCC2), the kidney-specific member of a cation-chloride cotransporter superfamily, is an integral membrane protein responsible for the transepithelial reabsorption of NaCl. The role of NKCC2 is essential for renal volume regulation. Vasopressin (AVP) controls NKCC2 surface expression in cells of the thick ascending limb of the loop of Henle (TAL). We found that 40-70% of Triton X-100-insoluble NKCC2 was present in cholesterol-enriched lipid rafts (LR) in rat kidney and cultured TAL cells. The related Na(+)-Cl(-) cotransporter (NCC) from rat kidney was distributed in LR as well. NKCC2-containing LR were detected both intracellularly and in the plasma membrane. Bumetanide-sensitive transport of NKCC2 as analyzed by (86)Rb(+) influx in Xenopus laevis oocytes was markedly reduced by methyl-beta-cyclodextrin (MbetaCD)-induced cholesterol depletion. In TAL, short-term AVP application induced apical vesicular trafficking along with a shift of NKCC2 from non-raft to LR fractions. In parallel, increased colocalization of NKCC2 with the LR ganglioside GM1 and their polar translocation were assessed by confocal analysis. Apical biotinylation showed twofold increases in NKCC2 surface expression. These effects were blunted by mevalonate-lovastatin/MbetaCD-induced cholesterol deprivation. Collectively, these findings demonstrate that a pool of NKCC2 distributes in rafts. Results are consistent with a model in which LR mediate polar insertion, activity, and AVP-induced trafficking of NKCC2 in the control of transepithelial NaCl transport.
AB - Apical bumetanide-sensitive Na(+)-K(+)-2Cl(-) cotransporter (NKCC2), the kidney-specific member of a cation-chloride cotransporter superfamily, is an integral membrane protein responsible for the transepithelial reabsorption of NaCl. The role of NKCC2 is essential for renal volume regulation. Vasopressin (AVP) controls NKCC2 surface expression in cells of the thick ascending limb of the loop of Henle (TAL). We found that 40-70% of Triton X-100-insoluble NKCC2 was present in cholesterol-enriched lipid rafts (LR) in rat kidney and cultured TAL cells. The related Na(+)-Cl(-) cotransporter (NCC) from rat kidney was distributed in LR as well. NKCC2-containing LR were detected both intracellularly and in the plasma membrane. Bumetanide-sensitive transport of NKCC2 as analyzed by (86)Rb(+) influx in Xenopus laevis oocytes was markedly reduced by methyl-beta-cyclodextrin (MbetaCD)-induced cholesterol depletion. In TAL, short-term AVP application induced apical vesicular trafficking along with a shift of NKCC2 from non-raft to LR fractions. In parallel, increased colocalization of NKCC2 with the LR ganglioside GM1 and their polar translocation were assessed by confocal analysis. Apical biotinylation showed twofold increases in NKCC2 surface expression. These effects were blunted by mevalonate-lovastatin/MbetaCD-induced cholesterol deprivation. Collectively, these findings demonstrate that a pool of NKCC2 distributes in rafts. Results are consistent with a model in which LR mediate polar insertion, activity, and AVP-induced trafficking of NKCC2 in the control of transepithelial NaCl transport.
KW - Animals
KW - Arginine Vasopressin
KW - Biotinylation
KW - Cell Polarity
KW - Cells, Cultured
KW - Cholesterol
KW - Chromatography, Liquid
KW - Diabetes Insipidus, Neurogenic
KW - Kidney
KW - Loop of Henle
KW - Male
KW - Mass Spectrometry
KW - Membrane Microdomains
KW - Oocytes
KW - Rabbits
KW - Rats
KW - Rats, Long-Evans
KW - Rats, Sprague-Dawley
KW - Receptors, Vasopressin
KW - Sodium-Potassium-Chloride Symporters
KW - Solute Carrier Family 12, Member 1
KW - Xenopus
KW - Journal Article
KW - Research Support, N.I.H., Extramural
KW - Research Support, Non-U.S. Gov't
U2 - 10.1152/ajprenal.90227.2008
DO - 10.1152/ajprenal.90227.2008
M3 - SCORING: Journal article
C2 - 18579701
VL - 295
SP - 789
EP - 802
JO - AM J PHYSIOL-RENAL
JF - AM J PHYSIOL-RENAL
SN - 1931-857X
IS - 3
M1 - 3
ER -