Localization of a proton-pumping ATPase in rat kidney.

Brown, Dennis; Hirsch, Sheldon; Gluck, Stephen L.

doi:10.1172/jci113833

Cited by 318 publications

(211 citation statements)

References 51 publications

(61 reference statements)

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“…Co-localization of GFP with H ϩ -ATPase staining was also confirmed by double labeling, as shown in Figure 4F. H ϩ -ATPase was also expressed in the proximal convoluted tubule cells, where it localized to the base of the brush border as reported previously (28). Examination of serial sections revealed that proximal tubules with apical H ϩ -ATPase staining were not positive for GFP, while tubules that contained GFP positive cells were negative for H ϩ -ATPase.…”

Section: Immunohistochemical Detection Of Transgene Expressionsupporting

confidence: 85%

Gene Delivery in Renal Tubular Epithelial Cells Using Recombinant Adeno-Associated Viral Vectors

Chen¹,

Agarwal²,

Glushakova³

et al. 2003

Journal of the American Society of Nephrology

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ABSTRACT. Gene therapy has the potential to provide a therapeutic strategy for numerous renal diseases such as diabetic nephropathy, chronic rejection, Alport syndrome, polycystic kidney disease, and inherited tubular disorders. In previous studies using cationic liposomes or adenoviral or retroviral vectors to deliver genes into the kidney, transgene expression has been transient and often associated with adverse host immune responses, particularly with the use of adenoviral vectors. The unique properties of recombinant adeno-associated viral (rAAV) vectors permit long-term stable transgene expression with a relatively low host immune response. The purpose of the present study was to evaluate gene expression in the rat kidney after intrarenal arterial infusion of a rAAV (serotype 2) vector encoding green fluorescence protein (GFP) induced by a cytomegalovirus-chicken beta-actin hybrid promoter. The left kidney of experimental animals was treated with either saline or transduced with rAAV2-GFP (0.125 ml/100 g body wt, 1 × 1010/ml infectious units) through the renal artery. A time-dependent expression of GFP was observed in all kidneys injected with rAAV2-GFP, with maximal expression observed at 6 wk posttransduction. The expression of GFP was restricted to cells in the S3 segment of the proximal tubule and intercalated cells in the collecting duct, the latter identified by co-localization with H+-ATPase. No transduction was observed in the glomeruli or the intrarenal vasculature. These studies demonstrate successful transgene expression in tubular epithelial cells, specifically in the S3 segment of the proximal tubule and intercalated cells, after intrarenal administration of a rAAV vector and provide the impetus for further studies to exploit its use as a tool for gene therapy in the kidney.E-mail: agarwal@nersp.nerdc.ufl.edu

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Section: Immunohistochemical Detection Of Transgene Expressionsupporting

confidence: 85%

Gene Delivery in Renal Tubular Epithelial Cells Using Recombinant Adeno-Associated Viral Vectors

Chen¹,

Agarwal²,

Glushakova³

et al. 2003

Journal of the American Society of Nephrology

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“…The presence of N-ethylmaleimide (NEM)-sensitive vacuolar H + -ATPase in the TALs has been demonstrated first in rat kidneys (Brown et al 1988). The H + -ATPase is selectively localized to luminal membranes and submembranous vesicles.…”

Section: Relation To H + -Atpase and Na + /H + Exchangers In The Lumimentioning

confidence: 99%

Chloride-Dependent Intracellular pH Regulation via Extracellular Calcium-Sensing Receptor in the Medullary Thick Ascending Limb of the Mouse Kidney

Aslanova

Morimoto

Farajov

et al. 2006

Tohoku J. Exp. Med.

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The extracellular calcium-sensing receptor (CaSR) located in either luminal or basolateral cell membranes of various types of renal tubules including proximal tubules, Henle's loop and collecting ducts has been thought to play a fundamental role in electrolyte metabolism. To further identify the physiological roles of the CaSR, we examined the effects of Ca 2+ and calcimimetics neomycin (Neo), gentamicin and gadolinium chloride (Gd 3+ ) on the intracellular pH (pHi) of in vitro microperfused mouse medullary thick ascending limb (mTAL) cells of Henle's loop, by loading the cells with fluorescent pH indicator 2′,7′-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein and measuring the ratio of fluorescence emission at 530 nm after exciting the dye at 490 and 440 nm. In a steady-state condition in Hepes-buffered solution, the pHi in the mTALs was 7.29 ± 0.04 (n = 9). A concentration of 200 μ mol/l Neo in the basolateral side decreased the pHi after 1 min by −0.13 ± 0.02 (n = 34, p < 0.0001). The other calcimimetics showed similar effects on pHi, whereas none of these calcimimetics in the lumen affected pHi. Na + removal or the inhibition of Na + and proton transport with amiloride, bumetanide, or bafilomycin did not eliminate the effect of Neo on pHi. On the other hand, Cl − removal clearly eliminated the Neo-induced pHi decrease (−0.06 ± 0.01 vs −0.00 ± 0.05 in Cl − removal, n = 4, p < 0.003). Thus, we have demonstrated for the first time that the CaSR is involved in the regulation of the pHi in the mTAL and requires Cl − to exert its effect. intracellular pH; renal medulla; chloride; amiloride; BCECF

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“…Rat. Antibodies against B1 subunit isoform (29) were used followed by protein A conjugated to 8-nm colloidal gold particles as described (30). Briefly, 1-m sections picked up on nickel grids were processed with no prior etching procedure.…”

Section: Fig 1 Schematic Representation Of the Ocular Ciliary Epithmentioning

confidence: 99%

Vacuolar H ⁺ -ATPase in ocular ciliary epithelium

Wax

Saito

Tenkova

et al. 1997

Proc. Natl. Acad. Sci. U.S.A.

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The mechanisms controlling the production of aqueous humor and the regulation of intraocular pressure are poorly understood. Here, we provide evidence that a vacuolar H ؉ -ATPase (V-ATPase) in the ocular ciliary epithelium is a key component of this process. In intracellular pH (pH i ) measurements of isolated ciliary epithelium performed with 2 ,7-biscarboxyethyl-5(6)-carboxyf luorescein (BCECF), the selective V-ATPase inhibitor bafilomycin A 1 slowed the recovery of pH i in response to acute intracellular acidification, demonstrating the presence of V-ATPase in the plasma membrane. In isolated rabbit ciliary body preparations examined under voltage-clamped conditions, bafilomycin A 1 produced a concentration-dependent decrease in short-circuit current, and topical application of bafilomycin A 1 reduced intraocular pressure in rabbits, indicating an essential role of the VATPase in ciliary epithelial ion transport. Immunocytochemistry utilizing antibodies specific for the B1 isoform of the V-ATPase 56-kDa subunit revealed localization of V-ATPase in both the plasma membrane and cytoplasm of the native ciliary epithelium in both rabbit and rat eye. The regional and subcellular distribution of V-ATPase in specific regions of the ciliary process was altered profoundly by isoproterenol and phorbol esters, suggesting that change in the intracellular distribution of the enzyme is a mechanism by which drugs, hormones, and neurotransmitters modify aqueous humor production.

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Localization of a proton-pumping ATPase in rat kidney.

Cited by 318 publications

References 51 publications

Gene Delivery in Renal Tubular Epithelial Cells Using Recombinant Adeno-Associated Viral Vectors

Gene Delivery in Renal Tubular Epithelial Cells Using Recombinant Adeno-Associated Viral Vectors

Chloride-Dependent Intracellular pH Regulation via Extracellular Calcium-Sensing Receptor in the Medullary Thick Ascending Limb of the Mouse Kidney

Vacuolar H ⁺ -ATPase in ocular ciliary epithelium

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Localization of a proton-pumping ATPase in rat kidney.

Cited by 318 publications

References 51 publications

Gene Delivery in Renal Tubular Epithelial Cells Using Recombinant Adeno-Associated Viral Vectors

Gene Delivery in Renal Tubular Epithelial Cells Using Recombinant Adeno-Associated Viral Vectors

Chloride-Dependent Intracellular pH Regulation via Extracellular Calcium-Sensing Receptor in the Medullary Thick Ascending Limb of the Mouse Kidney

Vacuolar H + -ATPase in ocular ciliary epithelium

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Vacuolar H ⁺ -ATPase in ocular ciliary epithelium