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H+ Exchange (h+ + exchange)
Selected AbstractsProtective Effect of HOE642, a Selective Blocker of Na+ -H+ Exchange, Against the Development of Rigor Contracture in Rat Ventricular MyocytesEXPERIMENTAL PHYSIOLOGY, Issue 1 2000Marisol Ruiz-Meana The objective of this study was to investigate the effect of Na+ -H+ exchange (NHE) and HCO3, -Na+ symport inhibition on the development of rigor contracture. Freshly isolated adult rat cardiomyocytes were subjected to 60 min metabolic inhibition (MI) and 5 min re-energization (Rx). The effects of perfusion of HCO3, or HCO3, -free buffer with or without the NHE inhibitor HOE642 (7 ,M) were investigated during MI and Rx. In HCO3, -free conditions, HOE642 reduced the percentage of cells developing rigor during MI from 79 ± 1% to 40 ± 4% (P < 0.001) without modifying the time at which rigor appeared. This resulted in a 30% reduction of hypercontracture during Rx (P < 0.01). The presence of HCO3, abolished the protective effect of HOE642 against rigor. Cells that had developed rigor underwent hypercontracture during Rx independently of treatment allocation. Ratiofluorescence measurement demonstrated that the rise in cytosolic Ca2+ (fura-2) occurred only after the onset of rigor, and was not influenced by HOE642. NHE inhibition did not modify Na+ rise (SBFI) during MI, but exaggerated the initial fall of intracellular pH (BCEFC). In conclusion, HOE642 has a protective effect against rigor during energy deprivation, but only when HCO3, -dependent transporters are inhibited. This effect is independent of changes in cytosolic Na+ or Ca2+ concentrations. [source] Cell shrinkage evoked by Ca2+ -free solution in rat alveolar type II cells: Ca2+ regulation of Na+,H+ exchangeEXPERIMENTAL PHYSIOLOGY, Issue 2 2005Hitoshi Murao The effects of intracellular Ca2+ concentration, [Ca2+]i, on the volume of rat alveolar type II cells (AT-II cells) were examined. Perfusion with a Ca2+ -free solution induced shrinkage of the AT-II cell volume in the absence or presence of amiloride (1 ,m, an inhibitor of Na+ channels); however, it did not in the presence of 5-(N -methyl- N -isobutyl)-amiloride (MIA, an inhibitor of Na+,H+ exchange). MIA decreased the volume of AT-II cells. Inhibitors of Cl,,HCO3, exchange, 4,4,-diisothiocyanostilbene-2,2,-disulfonic acid (DIDS) and 4-acetamido-4,-isothiocyanatostilbene-2,2,-disulfonic acid (SITS) also decreased the volume of AT-II cells. This indicates that the cell shrinkage induced by a Ca2+ -free solution is caused by a decrease in NaCl influx via Na+,H+ exchange and Cl,,HCO3, exchange. Addition of ionomycin (1 ,m), in contrast, induced cell swelling when AT-II cells were pretreated with quinine and amiloride. This swelling of the AT-II cells is not detected in the presence of MIA. Intracellular pH (pHi) measurements demonstrated that the Ca2+ -free solution or MIA decreases pHi, and that ionomycin increases it. Ionomycin stimulated the pHi recovery after an acid loading (NH4+ pulse method), which was not noted in MIA-treated AT-II cells. Ionomycin increased [Ca2+]i in fura-2-loaded AT-II cells. In conclusion, the Na+,H+ exchange activities of AT-II cells, which maintain the volume and pHi, are regulated by [Ca2+]i. [source] Protective Effect of HOE642, a Selective Blocker of Na+ -H+ Exchange, Against the Development of Rigor Contracture in Rat Ventricular MyocytesEXPERIMENTAL PHYSIOLOGY, Issue 1 2000Marisol Ruiz-Meana The objective of this study was to investigate the effect of Na+ -H+ exchange (NHE) and HCO3, -Na+ symport inhibition on the development of rigor contracture. Freshly isolated adult rat cardiomyocytes were subjected to 60 min metabolic inhibition (MI) and 5 min re-energization (Rx). The effects of perfusion of HCO3, or HCO3, -free buffer with or without the NHE inhibitor HOE642 (7 ,M) were investigated during MI and Rx. In HCO3, -free conditions, HOE642 reduced the percentage of cells developing rigor during MI from 79 ± 1% to 40 ± 4% (P < 0.001) without modifying the time at which rigor appeared. This resulted in a 30% reduction of hypercontracture during Rx (P < 0.01). The presence of HCO3, abolished the protective effect of HOE642 against rigor. Cells that had developed rigor underwent hypercontracture during Rx independently of treatment allocation. Ratiofluorescence measurement demonstrated that the rise in cytosolic Ca2+ (fura-2) occurred only after the onset of rigor, and was not influenced by HOE642. NHE inhibition did not modify Na+ rise (SBFI) during MI, but exaggerated the initial fall of intracellular pH (BCEFC). In conclusion, HOE642 has a protective effect against rigor during energy deprivation, but only when HCO3, -dependent transporters are inhibited. This effect is independent of changes in cytosolic Na+ or Ca2+ concentrations. [source] Modulation of Na+ -H+ exchange isoforms NHE1 and NHE3 by insulin-like growth factor-1 in isolated bovine articular chondrocytesJOURNAL OF ORTHOPAEDIC RESEARCH, Issue 11 2008Amanda L. Tattersall Abstract Incubation with serum modulates the transporters that regulate intracellular pH (pHi) in articular chondrocytes, upregulating acid extrusion by Na+ -H+ exchange (NHE). There is stimulation of NHE1, together with induction of NHE3 activity. These isoforms exhibit differential responses to components of mechanical load experienced by chondrocytes during joint loading. The identity of the component(s) of serum responsible is unknown. A possibility, however, is insulin-like growth factor-1 (IGF-1), present in normal cartilage and found at enhanced levels in osteoarthritic tissue. In the present study, the effects of IGF-1 on pHi regulation have been characterized using fluorescence measurements of bovine articular chondrocytes, and the sensitivity of pHi regulation to hyperosmotic shock and raised hydrostatic pressure determined. For cells isolated in the absence of IGF-1, pHi recovery following acidification was predominantly mediated by NHE1. Recovery was enhanced when cells were incubated for 18 h with 20 ng mL,1 IGF; this effect represented increased acid extrusion by NHE1, supplemented by NHE3 activity. NHE3 activity was not detected in IGF-1-treated cells that had been incubated with the protein synthesis inhibitor cycloheximide, although NHE1 activity was unaffected. In the absence of IGF-1, suspension in hyperosmotic solutions or raised hydrostatic pressure enhanced pHi recovery of acidified cells. This response was missing in cells incubated with IGF-1. Unresponsiveness to hyperosmotic shock represented inhibition of NHE3 activity, and was prevented using the protein kinase A inhibitor KT5720. For raised hydrostatic pressure, a decrease in NHE1 activity was responsible, and was prevented by the protein kinase C inhibitor chelerythrine. © 2008 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 26:1428,1433, 2008 [source] Voltage-dependent and -independent titration of specific residues accounts for complex gating of a ClC chloride channel by extracellular protonsTHE JOURNAL OF PHYSIOLOGY, Issue 7 2009María Isabel Niemeyer The ClC transport protein family comprises both Cl, ion channel and H+/Cl, and H+/NO3, exchanger members. Structural studies on a bacterial ClC transporter reveal a pore obstructed at its external opening by a glutamate side-chain which acts as a gate for Cl, passage and in addition serves as a staging post for H+ exchange. This same conserved glutamate acts as a gate to regulate Cl, flow in ClC channels. The activity of ClC-2, a genuine Cl, channel, has a biphasic response to extracellular pH with activation by moderate acidification followed by abrupt channel closure at pH values lower than ,7. We have now investigated the molecular basis of this complex gating behaviour. First, we identify a sensor that couples extracellular acidification to complete closure of the channel. This is extracellularly-facing histidine 532 at the N-terminus of transmembrane helix Q whose neutralisation leads to channel closure in a cooperative manner. We go on to show that acidification-dependent activation of ClC-2 is voltage dependent and probably mediated by protonation of pore gate glutamate 207. Intracellular Cl, acts as a voltage-independent modulator, as though regulating the pKa of the protonatable residue. Our results suggest that voltage dependence of ClC-2 is given by hyperpolarisation-dependent penetration of protons from the extracellular side to neutralise the glutamate gate deep within the channel, which allows Cl, efflux. This is reminiscent of a partial exchanger cycle, suggesting that the ClC-2 channel evolved from its transporter counterparts. [source] Basolateral anion transport mechanisms underlying fluid secretion by mouse, rat and guinea-pig pancreatic ductsTHE JOURNAL OF PHYSIOLOGY, Issue 2 2004M. Paz Fernández-Salazar Fluid secretion by interlobular pancreatic ducts was determined by using video microscopy to measure the rate of swelling of isolated duct segments that had sealed following overnight culture. The aim was to compare the HCO3, requirement for secretin-evoked secretion in mouse, rat and guinea-pig pancreas. In mouse and rat ducts, fluid secretion could be evoked by 10 nm secretin and 5 ,m forskolin in the absence of extracellular HCO3,. In guinea-pig ducts, however, fluid secretion was totally dependent on HCO3,. Forskolin-stimulated fluid secretion by mouse and rat ducts in the absence of HCO3, was dependent on extracellular Cl, and was completely inhibited by bumetanide (30 ,m). It was therefore probably mediated by a basolateral Na+,K+,2Cl, cotransporter. In the presence of HCO3,, forskolin-stimulated fluid secretion was reduced ,40% by bumetanide, ,50% by inhibitors of basolateral HCO3, uptake (3 ,m EIPA and 500 ,m H2DIDS), and was totally abolished by simultaneous application of all three inhibitors. We conclude that the driving force for secretin-evoked fluid secretion by mouse and rat ducts is provided by parallel basolateral mechanisms: Na+,H+ exchange and Na+,HCO3, cotransport mediating HCO3, uptake, and Na+,K+,2Cl, cotransport mediating Cl, uptake. The absence or inactivity of the Cl, uptake pathway in the guinea-pig pancreatic ducts may help to account for the much higher concentrations of HCO3, secreted in this species. [source] | ||||||||||