Differential expression of KCNQ1 K+ channel in tubular cells of frog kidney.

The aim of this study was to evaluate KCNQ1 K+ channel expression in the frog kidney of Rana esculenta. KCNQ1 K+ channel, also known as KvLQT1, is the pore forming a-subunit of the IKs K+ channel, a delayed rectifier voltage-gated K+ channel, which has an important role in water and salt transport in the kidney and gastrointestinal tract. The expression of KCNQ1 K+ channel along tubular epithelium differs from species to species. In the present study the expression of KCNQ1 K+ channel in the frog kidney has been demonstrated by immunohistochemistry. The presence of KCNQ1 K+ channel was demonstrated in the epithelial cells of distal convoluted tubule and collecting duct. However, the pattern of expression of KCNQ1 K+ channel differs between distal convoluted tubules and collecting duct. All epithelial cells of distal convoluted tubules revealed basolateral expression of KCNQ1 K+ channel. On the contrary, only the single cells of collecting duct, probably intercalated cells, showed diffuse cell surface staining with antibodies against KCNQ1 K+ channel. These findings suggest that KCNQ1 K+ channel has cell-specific roles in renal potassium ion transport.

Excitotoxicity of lathyrus sativus neurotoxin in leech retzius neurons.

The effects of Lathyrus sativus neurotoxin were studied on the cell membrane potential and cellular cation composition in Retzius nerve cells of the leech Haemopis sanguisuga, with ion-selective microelectrodes using liquid ion-exchangers. Bath application of 10(-4) mol/l Lathyrus sativus neurotoxin for 3 min depolarized the cell membrane potential and decreased the input resistance of directly polarized membrane in Retzius neurons. At the same time the cellular Na+ activity increased and cellular K+ activity decreased with slow but complete recovery, while the intracellular Ca2+ concentration was not changed. Na+-free Ringer solutions inhibited the depolarizing effect of the neurotoxin on the cell membrane potential. Zero-Ca2+ Ringer solution or Ni2+-Ringer solution had no influence on the depolarizing effect of the neurotoxin on the cell membrane potential. It is obvious that the increase in membrane conductance and depolarization of the cell membrane potential are due to an influx of Na+ into the cell accompanied by an efflux of K+ from the cell.

Effects of cumene hydroperoxide on cellular cation composition in frog kidney proximal tubular cells.

Effects of cumene hydroperoxide were studied on the peritubular membrane potential and cellular cation composition in frog kidney proximal tubular cells. After perfusion of isolated frog kidneys for 30 min with 1.3x10(-4) mol l(-1) cumene hydroperoxide Ringer solution, the peritubular membrane potential gradually declined. The ouabain-like effects were demonstrated on cell Na and K activities after 1 h of perfusion with cumene hydroperoxide. The peritubular apparent transference number for potassium was decreased. Intracellular pH was not altered in the presence of cumene hydroperoxide. Intracellular free Ca(2+) concentration increased slowly and moderately. The concentration of the malondialdehyde in the kidney homogenates, measured as an index of lipid peroxidation, was increased. A previously observable effect of cumene hydroperoxide on the peritubular membrane potential was prevented by oxygen radical scavengers.

Peritubular Na-K exchange ion pump in maleate-treated frog kidney proximal tubular cells.

1. After perfusion of isolated frog kidneys for 1 hr with 10(-3) or 10(-2) M maleate Ringer, the peritubular membrane potential gradually declined in a dose-dependent manner. 2. The ouabain-like effects of maleate on cell Na and K activities were dose-dependent and smaller than the effects of zero K or 10(-4) M ouabain. Intracellular pH was not altered in the presence of 10(-2) M maleate. 3. The driving force for Na entry into the cell was reduced, respectively, to 81.4 and 58.4% (of control) in the presence of 10(-3) and 10(-2) M maleate. 4. There was no histochemically detectable inhibition of proximal tubule Na-K ATPase activity during 3 hr of perfusion with 10(-2) M maleate.

Substrate activation of mechanosensitive, whole cell currents in renal proximal tubule.

Isolated, polarized, proximal tubule cells of Rana pipiens were voltage clamped and examined for both single-channel and whole cell currents. Barium-sensitive whole cell conductances were calculated from the difference in slopes of the current-voltage relations before and after 5 mM external barium. In 11 voltage-clamped cells with high K in the pipette (and cell), isosmotic addition of 40 mM glucose to the bathing solution increased cell volume by 23 +/- 4% within 2-3 min and increased barium-sensitive conductance by 40 +/- 10% from 0.5 to 0.7 nS (P < 0.005, with each cell as its own control). Isosmotic addition of nonmetabolizable methyl-alpha-D-glucopyranoside, which enters with Na across the apical membrane, produced a similar increase in barium-sensitive conductance (30 +/- 13%). In contrast, 3-O-methyl-D-glucopyranose, which is not cotransported with Na, did not alter either cell volume or barium-sensitive conductance. Isosmotic addition of 40 mM phenylalanine (Phe) increased cell volume by 21 +/- 3% and increased barium-sensitive conductance by 36 +/- 19% from 1.1 to 1.5 nS (P < 0.005, with each cell as its own control; n = 8). All K channels observed at the basolateral membrane of these amphibian cells were found to be activated by pipette suction (stretch) and inhibited by 5 mM external barium (outside-out patches). Hence, stretch-activated (SA) K channels must be mediating the macroscopic increase in whole cell K conductance (GK) after isosmotic addition of glucose and Phe. The process does not seem to involve changes in ATP, because Phe increased GK even more when cytosolic ATP was maintained at high levels (10(-4) M extracellular ouabain and 5 mM intracellular ATP). It is also unlikely that changes in cell pH or calcium mediate the increase in GK, because the bulk composition of the cell is "clamped" by the pipette solution in these experiments (1-micron tip patch pipettes). Consequently, the substrate-induced increase in GK probably arises from a swelling-associated deformation of the submembane cytoskeleton or a direct change in membrane tension. In either case, SA channels would play a physiological role in proximal tubule K homeostasis during sugar and amino acid reabsorption in the proximal tubule of the kidney.

Effects of acid Ca2+ Ringer on passive electrical properties and intracellular ion activities in leech Retzius neuron.

1. A significant drop in effective input resistance of the free membrane and an increase in effective coupling resistance in acid Ca2+ Ringer (complete replacement of Na+ with Ca2+, pH 4) compared to control medium has been obtained in leech Retzius neurons. 2. In neutral Ca2+ Ringer (pH 7.2), effective input resistance increased while effective coupling resistance did not change. In acid sodium, leech Ringer (pH 4) effective input resistance increased while coupling resistance decreased. 3. Ten millimolar manganese and 10 mmol tetraethylammonium did not block conductance changes obtained in acid Ca2+ Ringer. 4. Intracellular activity of Na+ decreased, cellular activity of Cl- increased and intracellular K+ activity was unchanged in both acid and neutral Ca2+ Ringer. 5. The main difference was intracellular acidification in acid Ca2+ Ringer while intracellular pH was unchanged in neutral Ca2+ Ringer. 6. We discuss the possibility that in acid Ca2+ Ringer, intracellular acidification in leech neurons may be responsible for accompanying conductive changes.

Intracellular potential and K+ activity in rat kidney proximal tubular cells in acidosis and K+ depletion.

Techniques were developed for the measurement of intracellular potentials and potassium activities in rat proximal tubule cells using double barreled K+ liquid-ion-exchanger microelectrodes. After obtaining measurements of stable and reliable control values, the effects of K+ depletion and metabolic and respiratory acidosis on the intracellular potential and K+ activity in rat kidney proximal tubular cells were determined. At a peritubular membrane potential of -66.3 +/- 1.3 mV (mean +/- SE), intracellular K+ activity was 65.9 +/- 2.0 mEq/liter in the control rats. In metabolic acidosis [70 mg NH4Cl/100 g body wt) the peritubular membrane potential was significantly reduced to -47.5 +/- 1.9 mV, and cellular K+ activity to 53.5 +/- 2.0 mEq/liter. In contrast, in respiratory acidosis (15% CO2) the peritubular membrane potential was significantly lowered to -46.1 +/- 1.39 mV, but the cellular K+ activity was maintained at an almost unchanged level of 63.7 +/- 1.9 mEq/liter. In K+ depleted animals (6 weeks on low K+ diet), the peritubular membrane potential was significantly higher than in control animals, -74.8 +/- 2.1 mV, and cellular K+ activity was moderately but significantly reduced to 58.1 +/- 2.7 mEq/liter, Under all conditions studied, cellular K+ was above electrochemical equilibrium. Consequently, an active mechanism for cellular K+ accumulation must exist at one or both cell membranes. Furthermore, peritubular HCO3- appears to be an important factor in maintaining normal K+ distribution across the basolateral cell membrane.

Plasma renin activity and hypertrophy of the right ventricle in hypoxic rats.

A NaCl load in chronically hypoxic rats abolished the increase in plasma renin activity occurring in rats exposed to hypoxia of the same degree and duration but with normal NaCl intake. The parallel reduction in hypoxic hypertrophy of the right ventricle in NaCl-loaded rats could be considered as indirect evidence supporting the view that renin may be involved in the development of heart hypertrophy.