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10.1152/ajprenal.00268.2012 http://scihub22266oqcxt.onion/10.1152/ajprenal.00268.2012 22674028!3468494!22674028     free     free     free Warning: file_get_contents(https://eutils.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&id=22674028&cmd=llinks): Failed to open stream: HTTP request failed! 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Membrane-associated aquaporin-1 facilitates osmotically driven water flux across the basolateral membrane of the thick ascending limb |pdf=|usr=}}{{22674028}} gab.com Text Membrane-associated aquaporin-1 facilitates osmotically driven water flux across the basolateral membrane of the thick ascending limb JO: Am J Physiol Renal Physiol http://www.kidney.de/pget.php?&tw=1&pmid=22674028 #FOAvip The thick ascending limb of the loop of Henle (TAL) reabsorbs approximately 30% of filtered NaCl but is impermeable to water. The observation that little water traverses the TAL indicates an absence of water channels at the apical membrane. Yet TAL cells swell when peritubular osmolality decreases indicating that water channels must be present in the basolateral side. Consequently, we hypothesized that the water channel aquaporin-1 (AQP1) facilitates water flux across the basolateral membrane of TALs. Western blotting revealed AQP1 expression in microdissected rat and mouse TALs. Double immunofluorescence showed that 95 +/- 2% of tubules positive for the TAL-specific marker Tamm-Horsfall protein were also positive for AQP1 (n = 6). RT-PCR was used to demonstrate presence of AQP1 mRNA and the TAL-specific marker NKCC2 in microdissected TALs. Cell surface biotinylation assays showed that 23 +/- 3% of the total pool of AQP1 was present at the TAL basolateral membrane (n = 7). To assess the functional importance of AQP1 in the basolateral membrane, we measured the rate of cell swelling initiated by decreasing peritubular osmolality as an indicator of water flux in microdissected TALs. Water flux was decreased by approximately 50% in Aqp1 knockout mice compared with wild-types (4.0 +/- 0.8 vs. 8.9 +/- 1.7 fluorescent U/s, P < 0.02; n = 7). Furthermore, arginine vasopressin increased TAL AQP1 expression by 135 +/- 17% (glycosylated) and 41 +/- 11% (nonglycosylated; P < 0.01; n =5). We conclude that 1) the TAL expresses AQP1, 2) approximately 23% of the total pool of AQP1 is localized to the basolateral membrane, 3) AQP1 mediates a significant portion of basolateral water flux, and 4) AQP1 is upregulated in TALs of rats infused with dDAVP. AQP1 could play an important role in regulation of TAL cell volume during changes in interstitial osmolality, such as during a high-salt diet or water deprivation. Twit Text FOAVip Membrane-associated aquaporin-1 facilitates osmotically driven water flux across the basolateral membrane of the thick ascending limb ????Am J Physiol Renal Physiol http://www.kidney.de/pget.php?&tw=1&pmid=22674028 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=22674028 #FOAvip English Wikipedia <ref>{{Cite pmid|22674028}}</ref> Membrane-associated aquaporin-1 facilitates osmotically driven water flux across the basolateral membrane of the thick ascending limb #MMPMID22674028 Cabral PD; Herrera M Am J Physiol Renal Physiol 2012[Sep]; 303 (5): F621-9 PMID22674028show ga The thick ascending limb of the loop of Henle (TAL) reabsorbs approximately 30% of filtered NaCl but is impermeable to water. The observation that little water traverses the TAL indicates an absence of water channels at the apical membrane. Yet TAL cells swell when peritubular osmolality decreases indicating that water channels must be present in the basolateral side. Consequently, we hypothesized that the water channel aquaporin-1 (AQP1) facilitates water flux across the basolateral membrane of TALs. Western blotting revealed AQP1 expression in microdissected rat and mouse TALs. Double immunofluorescence showed that 95 +/- 2% of tubules positive for the TAL-specific marker Tamm-Horsfall protein were also positive for AQP1 (n = 6). RT-PCR was used to demonstrate presence of AQP1 mRNA and the TAL-specific marker NKCC2 in microdissected TALs. Cell surface biotinylation assays showed that 23 +/- 3% of the total pool of AQP1 was present at the TAL basolateral membrane (n = 7). To assess the functional importance of AQP1 in the basolateral membrane, we measured the rate of cell swelling initiated by decreasing peritubular osmolality as an indicator of water flux in microdissected TALs. Water flux was decreased by approximately 50% in Aqp1 knockout mice compared with wild-types (4.0 +/- 0.8 vs. 8.9 +/- 1.7 fluorescent U/s, P < 0.02; n = 7). Furthermore, arginine vasopressin increased TAL AQP1 expression by 135 +/- 17% (glycosylated) and 41 +/- 11% (nonglycosylated; P < 0.01; n =5). We conclude that 1) the TAL expresses AQP1, 2) approximately 23% of the total pool of AQP1 is localized to the basolateral membrane, 3) AQP1 mediates a significant portion of basolateral water flux, and 4) AQP1 is upregulated in TALs of rats infused with dDAVP. AQP1 could play an important role in regulation of TAL cell volume during changes in interstitial osmolality, such as during a high-salt diet or water deprivation. |Animals[MESH] |Aquaporin 1/*metabolism[MESH] |Cell Membrane/metabolism[MESH] |Deamino Arginine Vasopressin/pharmacology[MESH] |Loop of Henle/drug effects/*metabolism[MESH] |Mice[MESH] |Mice, Knockout[MESH] |Rats[MESH] |Uromodulin[MESH]Membrane-associated aquaporin-1 facilitates osmotically driven water f(epmc).pdf DeepDyve Pubget Overpricing