Polarized Sorting of Aquaporins 5 and 8 in Stable MDCK-II Transfectants

Wellner, Robert B.; Baum, Bruce J.

doi:10.1006/bbrc.2001.5287

Cited by 20 publications

(17 citation statements)

References 33 publications

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“…This localization is different from that found in submandibular gland acinar cells [149]. The stable expression of AQP8 cDNA in MDCK cells indicates basolateral expression of the protein in cultured cells [146].…”

Section: Epithelial Cell Models For Studying Subcellular Localizationcontrasting

confidence: 75%

“…Wellner and Baum reported that AQP5 is constitutively expressed at the apical membrane of MDCK cells [146], which resembles localization in vivo. The same authors made a series of protein chimeras with other known aquaporins to identify the sorting signals responsible for AQP5 apical localization [147,148].…”

Section: Epithelial Cell Models For Studying Subcellular Localizationmentioning

confidence: 85%

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Cell culture models and animal models for studying the patho-physiological role of renal aquaporins

Tamma

Procino

Svelto

et al. 2011

Cell. Mol. Life Sci.

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Aquaporins (AQPs) are key players regulating urinary-concentrating ability. To date, eight aquaporins have been characterized and localized along the nephron, namely, AQP1 located in the proximal tubule, thin descending limb of Henle, and vasa recta; AQP2, AQP3 and AQP4 in collecting duct principal cells; AQP5 in intercalated cell type B; AQP6 in intercalated cells type A in the papilla; AQP7, AQP8 and AQP11 in the proximal tubule. AQP2, whose expression and cellular distribution is dependent on vasopressin stimulation, is involved in hereditary and acquired diseases affecting urine-concentrating mechanisms. Due to the lack of selective aquaporin inhibitors, the patho-physiological role of renal aquaporins has not yet been completely clarified, and despite extensive studies, several questions remain unanswered. Until the recent and large-scale development of genetic manipulation technology, which has led to the generation of transgenic mice models, our knowledge on renal aquaporin regulation was mainly based on in vitro studies with suitable renal cell models. Transgenic and knockout technology approaches are providing pivotal information on the role of aquaporins in health and disease. The main goal of this review is to update and summarize what we can learn from cell and animal models that will shed more light on our understanding of aquaporin-dependent renal water regulation.

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Section: Epithelial Cell Models For Studying Subcellular Localizationcontrasting

confidence: 75%

Section: Epithelial Cell Models For Studying Subcellular Localizationmentioning

confidence: 85%

Cell culture models and animal models for studying the patho-physiological role of renal aquaporins

Tamma

Procino

Svelto

et al. 2011

Cell. Mol. Life Sci.

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“…AQP5 and AQP8 expressed in MDCK-II cells by genetransfection were shown to be sorted to apical and basolateral membranes, respectively; the sorting of AQP5 to apical membranes appears to require a recognition of sorting machinery [37]. AQP8 apparently can be sorted to either apical or basolateral membrane depending upon the cell type.…”

Section: Discussionmentioning

confidence: 99%

Divergent expression and localization of aquaporin 5, an exocrine-type water channel, in the submandibular gland of Sprague-Dawley rats

Murdiastuti

Miki

Yao

et al. 2002

Pfl�gers Archiv European Journal of Physiology

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By Western blot analysis, the expression level of aquaporin (AQP) 5 in the submandibular gland (SMG) was found to be different among individual rats of the Sprague-Dawley (SD) strain. Such differences were observed for AQP5 but not for AQP1 and consequently the SD strain was divided into two groups, one expressing a high level of AQP5 and the other a low one. The difference in average intensity of expression between the two groups was more than twofold. Immunohistochemical analysis of the SMG demonstrated that the AQP5 protein was localized in the basal and apical/lateral plasma membrane of acinar cells in rats expressing the high level of AQP5. In the rat expressing the low level, however, this channel protein was localized strongly in the apical/lateral plasma membrane, but only very weakly in the basal membrane of the acinar cells. Such a diverse localization of AQP5 was confirmed by Western blotting as well. Breeding between brother and sister was repeated for two times within high expressers and low expressers to obtain the third generation progenies (F2); the AQP5 level of the SMG in the third generation (F2 rats) from high expressers was significantly higher than the F2 from low expressers. Our present study suggests the existence of genetic variation in the expression of a water channel protein, AQP5, in rats.

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“…The transfection medium was removed after 5.5 h of incubation and replaced with fresh DMEM containing 10% FBS without antibiotics. After cultivation for 24 h following transfection, in which time point MDCK II cells had been cultured for 3 days in total (22), polycarbonate membranes containing cell monolayers were washed with phosphate -buffered saline (PBS) 3 times, fixed with 3% paraformaldehyde for 20 min, washed, and incubated with 50 mM NH4Cl for 15 min. For biotinylation, cells were washed, and blocked with 1% BSA (fraction V) in PBS for 1 h at room tenperature and reacted with cell-impermeable biotinylation reagent (2 mM sulfo-NHS-LC-biotin) in PBS containing 1 mM MgCl2, 0.1 mM CaCl2, pH 7.4 at room temperature for 40 min.…”

Section: Cell Culture Transient Transfection Cell Surface Biotinylamentioning

confidence: 99%

“…Thus trafficking and/or translocation of particular proteins, including AQP5 protein (22), toward apical or basolateral membranes can be studied in this model system. MDCK-II cells were transfected with plasmids generating chimeric proteins for GFP-AQP5(wt) or GFP-AQP5 proteins with mutations in PKA consensus sequence, and cytoplasmic or apical-membrane localization of these gene products was analyzed (Figs.…”

Section: Aqp5 Expression In Mdck-ii Cellsmentioning

confidence: 99%

Trafficking of GFP-AQP5 chimeric proteins conferred with unphosphorylated amino acids at their PKA-target motif (152SRRTS) in MDCK-II cells

et al. 2009

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Polarized Sorting of Aquaporins 5 and 8 in Stable MDCK-II Transfectants

Cited by 20 publications

References 33 publications

Cell culture models and animal models for studying the patho-physiological role of renal aquaporins

Cell culture models and animal models for studying the patho-physiological role of renal aquaporins

Divergent expression and localization of aquaporin 5, an exocrine-type water channel, in the submandibular gland of Sprague-Dawley rats

Trafficking of GFP-AQP5 chimeric proteins conferred with unphosphorylated amino acids at their PKA-target motif (152SRRTS) in MDCK-II cells

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