Transepithelial transport of sodium and chloride ions in isolated skin of the frog,Rana esculenta L

Isolated frog skin, mounted in a Ussing apparatus, was investigated electrophysiologically. Application of amiloride, an inhibitor of sodium ion transport, and bumetanide, known to block the transport of chloride ions, revealed the effect of these ions on PD, both under control conditions and following mechanical stimulation. Under control conditions, mechanical stimulation of the skin caused hyperpolarization, i.e. a transient increase in the electrical potential difference. Preincubation in the presence of amiloride, or amiloride plus bumetanide, brought about both a decrease in electrical potential and an inhibition of the reaction upon stimulation. On the other hand, incubation with bumetanide resulted in a decrease in electrical potential, but did not affect the skin reaction after mechanical stimulation. The above results indicate that hyperpolarization of the frog skin following mechanical stimulation is caused by enhanced transepithelial transport of sodium ions which, in turn, is induced by stimulation of sensory receptors.     

Active transport of chloride across skin isolated from Rana esculenta]

The effect of SITS, ouabain and acetazolamide on potential difference (E) and short-circuit current (ccc) across isolated frog skin after Amiloride treatment have been investigated. It has been found that Amiloride (3.5 . 10(-5)M) reverses the E and ccc values. After Amiloride treatment, with the use of SITS (6 . 10(-4)M) positive values of E and ccc have been measured while Ouabain (10(-4)M) only slightly reduces the reversed E and ccc values. On the other hand the effect of Acetazolamide (6 . 10(-4)M) is additional to the effect of SITS. It is suggested that the frog skin possesses an active transport for Cl-, as demonstrated by the use of Amiloride, and that it is carried out by two distinct mechanism: the first SITS and Acetazolamide sensible, the second one sensible to Strofantine.     

Effect of furosemide on unidirectional fluxes of sodium and chloride across the skin of the frog, Rana pipiens.

1. The diuretic furosemide, when added to the outside solution at a concentration of 5-10-4 M, increases the electrical potential difference (PD) across the isolated frog skin, but the short-circuit current (Isc) is unchanged. Lower concentrations had no significant effect on these electrical parameters. 2. When SO42- or NO3- are substituted for Cl- in the Ringer's solution furosemide has no effect on the PD or Isc. 3. Simultaneous unidirectional fluxes of Na+ and Cl- show that furosemide (5-10-4 M outside) reduces both the influx and outflux of Cl-, while the Na+ fluxes are not altered. 4. Furosemide (5-10-4 M) on the corium side of the frog skin had no significant effect on either PD, Isc or undirectional fluxes of Cl-. 5. It is suggested that furosemide reduces passive Cl- transfer, possibly by interacting with the Cl-/Cl- exchange diffusion mechanism which has been observed in this tissue. These observations further suggest that perhaps the diuretic action of furosemide may be mediated by such an effect on passive Cl- permeability which is linked to the active Cl- transport mechanism in the renal tubule.     

Role of external calcium in sodium and chloride transport in the gills of seawater-adaptedMugil capito

Summary When the mulletMugil capito is transferred to medium lacking Ca⁺⁺ (either Ca⁺⁺-free seawater or distilled water) the passive permeability of the gill to Na⁺ and Cl^– is increased and the activating effect of external K⁺ on the Na⁺ and Cl^– effluxes in hyposaline media is inhibited. The permeability of the gill increases progressively in proportion to the time of Ca⁺⁺ deprivation; it declines when Ca⁺⁺ is added again to the external medium. The active mechanisms for ion excretion are not reversible. At external Ca⁺⁺ concentrations from 0.1 to 10 mM the Na⁺ permeability is constant but the activation of Na⁺ efflux by K⁺ shows a maximum at a Ca⁺⁺ concentration of about 1 mM. For activation of Cl^– efflux external bicarbonate must be present, in addition to Ca⁺⁺, suggesting the existence of a Cl^–/HCO ₃ ^– exchange. The mechanism by which Ca⁺⁺ controls the passive branchial permeability is thus probably different from that involved in K⁺ activation of ion excretion. The Ca⁺⁺ effect on the K⁺ sensitive ionic excretory mechanisms seems to be related to intracellular Ca⁺⁺ movements. Thus, on the one hand, substances such as Ruthenium Red and La⁺⁺⁺ which both inhibit Ca⁺⁺ exchange, in media containing Ca⁺⁺ and HCO ₃ ^– also inhibit K⁺ activation of Na⁺ and Cl^– effluxes; on the other hand, the ionophore A 23187, a stimulator of Ca⁺⁺ exchange, when added to these media, activates the Na⁺ and Cl^– effluxes; its maximal effect on the Na⁺ flux occurs at 2 mM Ca⁺⁺.Abbreviations ASW-Ca artificial seawater minus calcium - DW deionised water - DWCa deionised water with 1 mM Ca⁺⁺ added - DWCaHCO ₃ DW with calcium plus bicarbonate - DWHCO ₃ DW with 1 mM sodium bicarbonate added - FW freshwater (tap water) - FWK freshwater with K⁺ added - P. D. potential difference - SW seawater The experiments reported in this paper were done with Jean Maetz who tragically died in August 1977. It is the last report about several years of friendly collaboration     

Effect of hibernation on sodium and chloride ion transport in isolated frog skin

The aim of the study was to evaluate the effect of hibernation on electrophysiological parameters of isolated frog skin under control incubation (Ringer solution) and after inhibition of Na+ and CI- transepithelial transport by application of amiloride and bumetanide. The transepithelial electrical potential difference (PD in mV) was measured before and after mechanical stimulation of isolated frog skin. The tissues were mounted in a modified Ussing chamber. The results revealed a reduced PD of frog skin during hibernation. In February, as compared with November, PD of frog skin incubated in Ringer solution decreased by about 50%. Hibernation also affected hyperpolarization (dPD) of frog skin after mechanical stimulation. In November and December, dPD was about 50% and 30% lower, respectively, compared with the subsequent two months of the experiment. The incubation of frog skin with amiloride, a sodium ion channel blocker, resulted in reduced values of all measured electrophysiological parameters irrespective of the phase of hibernation. After application of chloride ion transport inhibitor (bumetanide), the PD in November and December decreased compared with the control incubation by about 80% and 75%, while in January and February by about 40% and 25%, respectively. In January and February dPD increased by four times and three times as compared with November and December. Hibernation reduces net ion flow in isolated frog skin. During the initial period of hibernation the sensitivity of the skin to mechanical stimulation also decreases. Towards the end of hibernation, on the other hand, excitation of mechanosensitive ion channels takes place.     

Renal handling of phosphate, calcium, sodium, and potassium in intact and parathyroidectomized Rana pipiens

Renal excretion of phosphate, calcium, sodium, and potassium in intact and parathyroidectomized male Rana pipiens was studied by renal clearance techniques using 14C-inulin. In intact frogs, 57% of filtered phosphate, 60% of filtered calcium, 97% of filtered sodium, and 89% of filtered potassium was reabsorbed by the renal tubules. Following parathyroidectomy, the rate of reabsorption of phosphate became significantly higher than that of the intact frog, and the relative phosphate clearance (fractional excretion) decreased. These changes corresponded with a gradual rise in serum phosphate values. There was no major effect on excretion patterns of calcium, sodium, or potassium after parathyroidectomy. These results suggest that in frogs the parathyroid glands strongly influence phosphate excretion patterns but have little effect on the excretion of calcium, sodium, or potassium.     

Effect of acetazolamide on sodium transport through the isolated frog skin at low and high external sodium concentrations.

The action of acetazolamine on sodium transport in $\text{Rana esculenta}$ skin was studied with the external face bathed in dilute (2mMM) or concentrated (Ringer) solutions of sodium chloride.The absorption of Na⁺ from a dilute solution is inhibited at an acetazolamide concentration of 10⁻⁵M. This is due to an inhibition of the influx: the efflux remains unchanged. Acetazolamide has no effect, however, on transport from Ringer solution.The graphic determination of the Na⁺ transport pool at the 2 mM NaCl concentration showed that acetazolamide diminished the pool without affecting the $\text{t}\text{1}\text{2}$. The inhibitor had no effect on the pool at the higher (Ringer) concentration.These results indicate that acetazolamide acts on the external barrier of the sodium transport compartment without affecting the active pump of this ion when it is being transported from a dilute sodium chloride solution.     

The interrelationships between sodium ion,calcium transport and oxygen utilization in the isolated chick chorioallantoic membrane

Summary The interrelationships between sodium ion, calcium transport and oxygen utilization have been investigated in the chick chorioallantoic membrane. The oxygen uptakes of the two surface layers of the tissue, the ectoderm and the endoderm, were separated into their basal, Na⁺ dependent and Ca⁺⁺ dependent components. The endoderm has a basal rate of respiration of 3.6 liters O₂/cm²/hr and a Na⁺ dependent component of 1.4 liters O₂/cm²/hr. The ectoderm has a basal rate of respiration of about 3.5 liters O₂/cm²/hr, and Na⁺ and Ca⁺⁺ dependent components of 1.1 and 3.6 liters O₂/cm²/hr, respectively. The rate of ectodermal calcium transport and calcium-stimulated oxygen uptake is strictly dependent on the presence of sodium in the bathing medium, and complex kinetics are observed as a function of sodium concentration. On the other hand, in 140mm Na⁺ the rate of calcium transport exhibits simple saturation kinetics as a function of calcium concentration. Ca⁺⁺/O₂ ratios determined for many different rates of transport give a ratio of about 0.5, a value much lower than similar ratios determined for other transport mechanisms. The calcium transport mechanism in the ectoderm responds to changes in transport rate very sluggishly, taking 30 to 50 min to give a maximum response. The differences between the calcium transport mechanism in this membrane and other known transport systems are discussed and it is suggested that these differences may represent the adaptations necessary for transcellular calcium transport.