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1.
Braz. j. med. biol. res ; 42(2): 155-163, Feb. 2009. ilus
Artigo em Inglês | LILACS | ID: lil-506881

RESUMO

Vacuolar H+-ATPase is a large multi-subunit protein that mediates ATP-driven vectorial H+ transport across the membranes. It is widely distributed and present in virtually all eukaryotic cells in intracellular membranes or in the plasma membrane of specialized cells. In subcellular organelles, ATPase is responsible for the acidification of the vesicular interior, which requires an intraorganellar acidic pH to maintain optimal enzyme activity. Control of vacuolar H+-ATPase depends on the potential difference across the membrane in which the proton ATPase is inserted. Since the transport performed by H+-ATPase is electrogenic, translocation of H+-ions across the membranes by the pump creates a lumen-positive voltage in the absence of a neutralizing current, generating an electrochemical potential gradient that limits the activity of H+-ATPase. In many intracellular organelles and cell plasma membranes, this potential difference established by the ATPase gradient is normally dissipated by a parallel and passive Cl- movement, which provides an electric shunt compensating for the positive charge transferred by the pump. The underlying mechanisms for the differences in the requirement for chloride by different tissues have not yet been adequately identified, and there is still some controversy as to the molecular identity of the associated Cl--conducting proteins. Several candidates have been identified: the ClC family members, which may or may not mediate nCl-/H+ exchange, and the cystic fibrosis transmembrane conductance regulator. In this review, we discuss some tissues where the association between H+-ATPase and chloride channels has been demonstrated and plays a relevant physiologic role.


Assuntos
Animais , Membrana Celular/metabolismo , Canais de Cloreto/metabolismo , ATPases Vacuolares Próton-Translocadoras/metabolismo , Osso e Ossos/enzimologia , Sistema Nervoso Central/enzimologia , Canais de Cloreto/fisiologia , Rim/enzimologia , Fígado/enzimologia , ATPases Vacuolares Próton-Translocadoras/fisiologia
2.
Braz. j. med. biol. res ; 40(5): 687-696, May 2007. graf, tab
Artigo em Inglês | LILACS | ID: lil-449084

RESUMO

The interaction between H+ extrusion via H+-ATPase and Cl- conductance was studied in the C11 clone of MDCK cells, akin to the intercalated cells of the collecting duct. Cell pH (pHi) was measured by fluorescence microscopy using the fluorescein-derived probe BCECF-AM. Control recovery rate measured after a 20 mM NH4Cl acid pulse was 0.136 ± 0.008 pH units/min (dpHi/dt) in Na+ Ringer and 0.032 ± 0.003 in the absence of Na+ (0 Na+). With 0 Na+ plus the Cl- channel inhibitor NPPB (10 æM), recovery was reduced to 0.014 ± 0.001 dpHi/dt. 8-Br-cAMP, known to activate CFTR Cl- channels, increased dpHi/dt in 0 Na+ to 0.061 ± 0.009 and also in the presence of 46 nM concanamycin and 50 æM Schering 28080. Since it is thought that the Cl- dependence of H+-ATPase might be due to its electrogenic nature and the establishment of a +PD (potential difference) across the cell membrane, the effect of 10 æM valinomycin at high (100 mM) K+ was tested in our cells. In Na+ Ringer, dpHi/dt was increased, but no effect was detected in 0 Na+ Ringer in the presence of NPPB, indicating that in intact C11 cells the effect of blocking Cl- channels on dpHi/dt was not due to an adverse electrical gradient. The effect of 100 æM ATP was studied in 0 Na+ Ringer solution; this treatment caused a significant inhibition of dpHi/dt, reversed by 50 æM Bapta. We have shown that H+-ATPase present in MDCK C11 cells depends on Cl- ions and their channels, being regulated by cAMP and ATP, but not by the electrical gradient established by electrogenic H+ transport.


Assuntos
Animais , Canais de Cloreto/metabolismo , ATPases Translocadoras de Prótons/metabolismo , Linhagem Celular , Células Clonais , Fluoresceínas , Corantes Fluorescentes , Concentração de Íons de Hidrogênio , Microscopia de Fluorescência
3.
Braz. j. med. biol. res ; 37(10): 1481-1489, Oct. 2004. tab, graf
Artigo em Inglês | LILACS | ID: lil-383041

RESUMO

In the present study we determined the effect of chronic diet supplementation with n-3 PUFA on renal function of healthy and cachectic subjects by providing fish oil (1 g/kg body weight) to female rats throughout pregnancy and lactation and then to their offspring post-weaning and examined its effect on renal function parameters during their adulthood. The animals were divided into four groups of 5-10 rats in each group: control, control supplemented with fish oil (P), cachectic Walker 256 tumor-bearing (W), and W supplemented with fish oil (WP). Food intake was significantly lower in the W group compared to control (12.66 ± 4.24 vs 25.30 ± 1.07 g/day). Treatment with fish oil significantly reversed this reduction (22.70 ± 2.94 g/day). Tumor growth rate was markedly reduced in the P group (16.41 ± 2.09 for WP vs 24.06 ± 2.64 g for W). WP group showed a significant increase in mean glomerular filtration rate compared to P and control (1.520 ± 0.214 ml min-1 kg body weight-1; P < 0.05). Tumor-bearing groups had low urine osmolality compared to control rats. The fractional sodium excretion decreased in the W group compared to control (0.43 ± 0.16 vs 2.99 ± 0.87 percent; P < 0.05), and partially recovered in the WP group (0.90 ± 0.20 percent). In summary, the chronic supplementation with fish oil used in this study increased the amount of fat in the diet by only 0.1 percent, but caused remarkable changes in tumor growth rate and cachexia, also showing a renoprotective function.


Assuntos
Animais , Masculino , Feminino , Gravidez , Ratos , Caquexia , Carcinoma 256 de Walker , Suplementos Nutricionais , Óleos de Peixe , Hipolipemiantes , Rim , Peso Corporal , Taxa de Filtração Glomerular , Ratos Wistar , Sódio
4.
ARBS annu. rev. biomed. sci ; 2: 5-37, 2000. ilus, graf
Artigo em Inglês | LILACS | ID: lil-296331

RESUMO

Cell homeostasis of H+ ions has been an object of wide interest in the last two decades, which has led to extended knowledge about a considerable number of membrane transport mechanisms responsible for keeping cell pH within physiological limits. Among these mechanisms the most important are Na+/H+ exchange, the vacuolar H+-ATPase, the H+-K+-ATPase, Cl-/HCO3 - exchange and Na+/HCO3 - cotransport. The present review covers both cellular function and molecular aspects of these transporters, starting from a discussion of the methods used for the determination of cell pH and epithelial H+ transport, and analysing their molecular constitution, cloning and known isoforms, as well as their functional role in the maintenance of cell pH and epithelial transport.


Assuntos
Animais , Trocadores de Sódio-Hidrogênio/fisiologia , Transporte Biológico , ATPase Trocadora de Hidrogênio-Potássio , Homeostase/fisiologia , Concentração de Íons de Hidrogênio , ATPases Translocadoras de Prótons , ATPase Trocadora de Sódio-Potássio/fisiologia
5.
Braz. j. med. biol. res ; 30(4): 479-86, Apr. 1997. graf
Artigo em Inglês | LILACS | ID: lil-191386

RESUMO

The present paper reviews work from our laboratories evaluating the importance of adrenal cortical hormones in acidification by proximal and cortical distal tubules. Proximal acidification was determined by stationary microperfusion, and measurement of bicarbonate reabsorption using luminal pH determination was performed with H+ -ion-sensitive microelectrodes. Rats were adrenalectomized (ADX) 48 h before the experiments, and corticosteroids (aldosterone(A), corticosterone(B), and 18-OH corticosterone (18-OH-B)) were injected intramuscularly 100 and 40 min before the experiments. In ADX rats stationary pH increased significantly to 7.03 as compared to sham-operated rats (6.78). Bicarbonate reabsorption decreased from 2.65 + 0.18 in sham-operated rats to 0.50 + 0.07 mmol cm-2 S(-1) after ADX. The administration of the three hormones stimulated proximal tubule acidification, reaching, however, only 47.2 per cent of the sham values in aldosterone-treated rats. Distal nephron acidification was studied by measuring urine minus blood pCO2 differences (U-B pCO2) in bicarbonate-loaded rats treated as above. This pCO2 difference is used as a measure of the distal nephron ability to secrete H+ ions into an alkaline urine. U-B pCO2 decreased significantly from 39.9 + 1.26 to 11.9 + 1.99 mmHg in ADX rats. When corticosteroids were given to ADX rats before the experiment, U-B pCO2 increased significantly, but reached control levels only when aldosterone (two 3-mug doses per rat) plus corticosterone (220 mug) were given together. In order to control for the effect of aldosterone on distal transepithelial potential difference one group of rats was treated with amiloride, which blocks distal sodium channels. Amiloride-treated rats still showed a significant reduction in U-B pCO2 after ADX. Only corticosterone and 18-OH-B but not aldosterone increased U-B pCO2 back to the levels of sham-operated rats. These results show that corticosteroids stimulate renal tubule acidification both in proximal and distal nephrons and provide some clues about the mechanism of action of these steroids.


Assuntos
Ratos , Animais , Aldosterona/metabolismo , Bicarbonatos/metabolismo , Pressão Sanguínea/fisiologia , Corticosterona/metabolismo , Néfrons/metabolismo , Potássio/metabolismo , Sódio/metabolismo , Corticosteroides/fisiologia , Adrenalectomia , Ratos Wistar
6.
Braz. j. med. biol. res ; 27(4): 831-850, Apr. 1994.
Artigo em Inglês | LILACS | ID: lil-319822

RESUMO

The present paper reviews mechanisms by which the kidney controls systemic acid-base balance, with emphasis on the role of the distal nephron, and particularly of the cortical distal tubule. These mechanisms are essentially based on H-ion transport along the whole nephron. In proximal tubule cells, approximately 80 of H-ion secretion is mediated by Na+/H+ exchange, and 20 by H(+)-ATPase. In the distal nephron, acid-base transport mechanisms are located mainly in intercalated cells. H-ion secretion is effected by vacuolar H(+)-ATPase in alpha-intercalated cells and, in K-depleted animals, also by the gastric type H/K ATPase. In animals in alkalosis, beta-intercalated cells secrete bicarbonate by an apical Cl-/HCO3- exchanger, while a basolateral H-ATPase transfers H-ions into the interstitium. In cortical distal tubule, these mechanisms have been shown to be present in the intercalated cells of the connecting segment and of the initial collecting duct (the late distal tubule of micropuncture experiments). In the convoluted distal tubule (early distal tubule), most H-ion secretion occurs by means of the Na+/H+ exchanger. These data show that the distal nephron, including the cortical distal tubule, is a nephron segment responsible for a sizeable portion of bicarbonate reabsorption and titratable acid generation, as well as for bicarbonate secretion under appropriate metabolic conditions, being therefore the site of fine regulation of renal mechanisms that maintain acid-base homeostasis.


Assuntos
Animais , Ratos , Equilíbrio Ácido-Base/fisiologia , Túbulos Renais Distais/fisiologia , Trocadores de Sódio-Hidrogênio/fisiologia , ATPases Translocadoras de Prótons/fisiologia , Bicarbonatos , Concentração de Íons de Hidrogênio , Néfrons/fisiologia , Ratos Brattleboro , Ratos Wistar
7.
Braz. j. med. biol. res ; 26(7): 773-7, Jul. 1993. tab, graf
Artigo em Inglês | LILACS | ID: lil-148731

RESUMO

To evaluate the relative importance of the V-type H(+)-ATPase in proximal bicarbonate reabsorption in vivo, proximal tubules of male and female Wistar rats (180 to 260 g) were perfused with bicarbonate-Ringer solution with and without the addition of 2 microM bafilomycin A1. Bafilomycin significantly increased stationary pH from 6.75 +/- 0.05 (N = 39) to 6.86 +/- 0.03 (N = 82), the stationary concentration of bicarbonate from 5.24 +/- 0.62 to 6.33 +/- 0.46 mM and the half-time of acidification from 3.72 +/- 0.22 to 4.65 +/- 0.25 s, and significantly decreased net bicarbonate reabsorption from 3.17 +/- 0.21 to 2.55 +/- 0.15 nmol s-1 cm-2, that is, by 20 per cent . Since bafilomycin is considered to be a specific inhibitor for V-type H(+)-ATPase, these data establish 1) the existence of this type of transport in the rat proximal tubule and 2) that approximately a fifth of the total proximal bicarbonate reabsorption is due to this mechanism of transport


Assuntos
Animais , Masculino , Feminino , Ratos , Antibacterianos/farmacologia , Bicarbonatos/farmacocinética , Túbulos Renais Proximais/metabolismo , Absorção , Transporte Biológico Ativo , Concentração de Íons de Hidrogênio , Eletrofisiologia , Potenciais da Membrana/efeitos dos fármacos , Perfusão , ATPases Translocadoras de Prótons/antagonistas & inibidores , ATPases Translocadoras de Prótons/metabolismo , Ratos Wistar , Túbulos Renais Proximais/enzimologia
8.
Braz. j. med. biol. res ; 26(7): 779-94, Jul. 1993. tab, graf
Artigo em Inglês | LILACS | ID: lil-148732

RESUMO

1. The cortical distal tubule of the rat kidney participates in the regulation of acid-base balance, showing bicarbonate reabsorption, secretion or absence of transport under different experimental conditions. In the present study, we measured differences in transepithelial pH using double ion-exchange resin/reference microelectrodes in control and alkalotic (chronic plus acute) male Wistar rats and in alkalotic rats receiving a K+ supplement in diet and infusion. 2. pH was measured in the tubule lumen during stationary microperfusion with 25 mM bicarbonate Ringer solution, and in peritubular vessels next to the perfused tubules. 3. Differences in transepithelial pH were 0.70 +/- 0.12 (N = 16) pH units in early distal tubules (ED) and 1.03 +/- 0.050 (N = 15) in late distal tubules LD) of control rats, 0.22 +/- 0.056 (N = 17) in ED and 0.25 +/- 0.050 (N = 20) in LD of alkalotic rats, and -0.02 +/- 0.039 (N = 24) in ED and -0.02 +/- 0.040 (N = 24) in LD of K(+)-supplemented alkalotic rats. 4. In control rats, the transepithelial potential difference (PD) (-8.9 +/- 1.45 mV (N = 16) in ED and -32.7 +/- 2.99 mV (N = 15) in LD) was not large enough to explain transepithelial H+ and HCO3- gradients, suggesting the presence of an active transport mechanism responsible for their maintenance. 5. The present data show that the cortical distal tubule is able to establish transepithelial pH (HCO3-) differences, that these differences are reduced by alkalosis and abolished by alkalosis plus K+ supplementation, and that, although inversion of pH gradients (evidence for bicarbonate secretion) was observed in individual tubules, this inversion was not significant in the groups studied


Assuntos
Animais , Masculino , Ratos , Alcalose/metabolismo , Túbulos Renais Distais/metabolismo , Bicarbonatos/metabolismo , Transporte Biológico Ativo , Eletrofisiologia , Epitélio/metabolismo , Equilíbrio Ácido-Base , Concentração de Íons de Hidrogênio , Microeletrodos , Perfusão , Potássio/metabolismo , Ratos Wistar
9.
Braz. j. med. biol. res ; 24(10): 1063-6, 1991. tab
Artigo em Inglês | LILACS | ID: lil-102090

RESUMO

To evaluate the effects of parathyroid hormone (PTH) on urinary acidification parameters, thyroparathyroidectomy was performed in normal (TPTX) and in calcium-supplemented rats (TPTX+Ca2**). Both groups were supplemented with thyroxin. Glomerular filtration rate (GFR) fell from 7.79 ñ 0.33 in the control group (C) to 4.88 ñ 0.26 ml min**-1Kg**-1 in TPTX, while net acid excretion fell from 5.65 ñ 0.22 in C to 3.76 ñ 0.26 µmol min**1Kg in TPTX. Kinetic dat of urinary acidification obtained by microperfusion techniques in proximal tubules showed that the half-time of acidification (t/2) rose from 4.75 ñ 0.24 s in C to 8.97 ñ 0.64s in TPTX and persisted elevated in TPTx +Ca**2+ (7.40 ñ 0.43s); in the latter group, stationary pH was not significantly different from that of the control group. Bicarbonate reabsorption (J**HCO3) fell from 2.18 ñ 0.15 in C to 0.823 ñ 0.082 in TPTX and was 1.53 ñ 0.073 nmol s**-1 cm**-2 in TPTX+Ca**2+. These suggest that normal pH gradients depend on normal calcium levels, but acidification half-times are dependent on PTH, which also contributes keeping glomerular hemodynamics and acidification rates at normal levels


Assuntos
Animais , Masculino , Ratos , Acidose/etiologia , Bicarbonatos/urina , Rim/fisiopatologia , Hormônio Paratireóideo/farmacologia , Ratos Endogâmicos BB
10.
Braz. j. med. biol. res ; 23(11): 1195-9, 1990. tab, ilus
Artigo em Inglês | LILACS | ID: lil-91626

RESUMO

The mechanism of proximal tubule potassium reabsorption was studied by stopped-flow microperfusion and determination of potassium activities by ion-sensitive resin microelectrodes. The proximal tubule was unable to establish transepithelial potassium gradients. perfusion with 20 mM K+ turned the lumen 3 mV more negative, an effect abolished by Ba2+. the half-time for K+ activities to reach their stationary level after perfusion with 1 mMK+ was significantly increased by Ba2+ from 4.25 ñ 0.14sto 11.0 ñ 1.71s, and aftr perfusion with 20 mMK+, from 5.43 ñ 0.20 to 12.53 ñ 0.90s. These data indicate that a significant fraction of potassium is transferred across proximal tubule spithelium by a transcellular, K+-channel-dependent route


Assuntos
Ratos , Animais , Potássio/farmacocinética , Túbulos Renais Proximais/metabolismo , Bário/farmacologia , Transporte Biológico , Canais de Potássio/fisiologia , Epitélio/metabolismo , Perfusão , Ratos Endogâmicos , Túbulos Renais Proximais
11.
Braz. j. med. biol. res ; 21(5): 1065-8, 1988. ilus
Artigo em Inglês | LILACS | ID: lil-63614

RESUMO

In order to study the mechanism of H-ion secretion in cortical distal tubules of the rat kedney, the luminal pH and transepithelial potential difference (PD) were measured with double-barrelled, pH-sensitive, resin/reference microelectrodes. perfusion of peritubular capillaries with low-sodium solutions increased luminal pH by 0.28 ñ 0.024 units. Perfusion of the lumen with 1 m§ amiloride increased luminal pH by 0.67 ñ 0.01 units. These changes could not be atributed to modification of transepithelial PD. We conclude that early distal acidification is sodium-dependent, possibly owing to the presence of Na+/H+ exchange


Assuntos
Ratos , Animais , Amilorida , Concentração de Íons de Hidrogênio , Sódio/metabolismo , Túbulos Renais Distais/fisiologia
16.
Arq. bras. cardiol ; 36(supl.1): 13-7, 1981.
Artigo em Português | LILACS | ID: lil-3342

RESUMO

Foi estudado o efeito do verapamil, um antagonista do calcio, sobre a funcao renal, empregando tecnicas de "clearance". Na primeira parte do trabalho, o verapamil foi infundido na arteria renal, na dose de 0,0025 mg/min obtendo-se acentuada queda da pressao arterial, de 116 a 54 mmHg, e significante aumento na excrecao fracional de calcio, de 0,464% para 5,37. Este aumento ocorreu independente de variacoes na carga filtrada e evidenciou um efeito da droga modificando as forcas de Starling no intersticio do tubulo proximal e diminuindo em consequencia a reabsorcao de eletrolitos e agua. Na segunda parte do trabalho, a droga foi infundida sistematicamente em dose similar. Houve reducao da pressao arterial media, com diminuicao significante do fluxo plasmatico renal (de 15,9 ml/min/Kg para 5,53) e do ritmo de filtracao glomerular (de 5,63 ml/min/Kg para 2,71). A vasoconstricao renal provocada pelo sistema simpatico, como resposta a queda da pressao arterial, desviaria o fluxo sanguineo para outras zonas mais vitais do organismo. A reabsorcao de eletrolitos e agua nao se modificou como consequencia provavel do efeito conjunto e oposto do aumento da fracao de filtracao e da queda do fluxo plasmatico pos-glomerular


Assuntos
Pressão Arterial , Cálcio , Taxa de Filtração Glomerular , Verapamil
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