Active Transport of Potassium and Oxygen Consumption in the Isolated Midgut of Hyalophora Cecropia

1967 ◽  
Vol 46 (2) ◽  
pp. 235-248
Author(s):  
W. R. HARVEY ◽  
J. A. HASKELL ◽  
K. ZERAHN
Keyword(s):  
Oxygen Consumption ◽  
Oxygen Uptake ◽  
Active Transport ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Na Transport ◽  
Maximum Value ◽  
Flux Measurements ◽  
Hyalophora Cecropia ◽  
K Transport

1. Flux measurements with 42K reveal that in the isolated midgut of Hyalophora cecropia 90 to 100 % of the short-circuit current is carried by the active transport of potassium from the blood-side to the lumen. 2. When K-transport is strongly depressed, either by withholding potassium from the blood side or by imposing a large positive potential on the lumen, the oxygen uptake of the isolated gut remains virtually unchanged. If the K-transport were to be energized by the negligible increase in oxygen uptake about 40 µ-equiv. of potassium would have to be transported for every µ-equiv. of extra oxygen taken up. This ratio of K-transport to oxygen uptake is thermodynamically impossible. 3. The ratio of potassium transported to total oxygen consumed when the midgut is bathed with 32 mM potassium on both sides is about 1.3 at temperatures of 25° and 15° C. The ratio must be smaller at lower potassium concentrations and is 2.0 at 73.5 mM-K, which may be approaching the maximum value. 4. Although the oxygen uptake is independent of the K-transport, the reverse is not true. There is a close dependency of K-transport on oxygen consumption. 5. K-transport by the midgut contrasts with Na-transport by the frog skin because Na-transport stimulates oxidative metabolism whereas K-transport does not. Evidently the coupling of transport to energy supply is different in the two systems.

scholarly journals Na+ and K+ transport at basolateral membranes of epithelial cells. I. Stoichiometry of the Na,K-ATPase.

1986 ◽  
Vol 87 (3) ◽  
pp. 467-483 ◽  
Author(s):  
T C Cox ◽  
S I Helman
Keyword(s):  
Short Circuit ◽  
Basolateral Membrane ◽  
Short Circuit Current ◽  
Ringer Solution ◽  
Na Transport ◽  
Basolateral Membranes ◽  
Dependent Component ◽  
Epithelial Sheets ◽  
Apical Membranes ◽  
K Transport

The stoichiometry of pump-mediated Na/K exchange was studied in isolated epithelial sheets of frog skin. 42K influx across basolateral membranes was measured with tissues in a steady state and incubated in either beakers or in chambers. The short-circuit current provided estimates of Na+ influx at the apical membranes of the cells. 42K influx of tissues bathed in Cl- or SO4-Ringer solution averaged approximately 8 microA/cm2. Ouabain inhibited 94% of the 42K influx. Furosemide was without effect on pre-ouabain-treated tissues but inhibited a ouabain-induced and Cl--dependent component of 42K influx. After taking into account the contribution of the Na+ load to the pump by way of basolateral membrane recycling of Na+, the stoichiometry was found to increase from approximately 2 to 6 as the pump-mediated Na+ transport rate increased from 10 to 70 microA/cm2. Extrapolation of the data to low rates of Na+ transport (less than 10 microA/cm2) indicated that the stoichiometry would be in the vicinity of 3:2. As pump-mediated K+ influx saturates with increasing rates of Na+ transport, Na+ efflux cannot be obligatorily coupled to K+ influx at all rates of transepithelial Na+ transport. These results are similar to those of Mullins and Brinley (1969. Journal of General Physiology. 53:504-740) in studies of the squid axon.

Ca-sensitive Na transport in sheep omasum

1999 ◽  
Vol 276 (6) ◽  
pp. G1331-G1344 ◽  
Author(s):  
Gerhard Schultheiss ◽  
Holger Martens
Keyword(s):  
Divalent Cations ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Voltage Divider ◽  
Close Correlation ◽  
Na Transport ◽  
Luminal Side ◽  
Flux Measurements ◽  
Voltage Divider Ratio ◽  
Peak Value

Na transport across a preparation of sheep omasum was studied. All tissues exhibited a serosa-positive short-circuit current ( I sc), with a range of 1–4 μeq ⋅ h−1 ⋅ cm−2. A Michaelis-Menten-type kinetic was found between the Na concentration and the I sc(Michaelis-Menten constant for transport of Na = 6.7 mM; maximal transport capacity of Na = 4.16 μeq ⋅ h−1 ⋅ cm−2). Mucosal amiloride (1 mM), phenamil (1 or 10 μ), or serosal aldosterone (1 μM for 6 h) did not change I sc. Removal of divalent cations (Ca and Mg) enhanced I sc considerably from 2.61 ± 0.24 to a peak value of 11.18 ± 1.1 μeq ⋅ h−1 ⋅ cm−2. The peak I sc(overshoot) immediately declined to a plateau I sc of ∼6–7 μeq ⋅ h−1 ⋅ cm−2. Na flux measurements showed a close correlation between changes in I sc and Na transport. Transepithelial studies demonstrated that K, Cs, Rb, and Li are transported, indicating putative nonselective cation channels, which are inhibited by divalent cations (including Ca, Mg, Sr, Ba) and by (trivalent) La. Intracellular microelectrode recordings from the luminal side clearly showed changes of voltage divider ratio when mucosal divalent cations were removed. The obtained data support the assumption of a distinct electrogenic Na transport mechanism in sheep omasum.

The Source of Short-Circuit Current Across Locust Rectum

1978 ◽  
Vol 77 (1) ◽  
pp. 107-122 ◽  
Author(s):  
D. WILLIAMS ◽  
J. E. PHILLIPS ◽  
W. T. PRINCE ◽  
J. MEREDITH
Keyword(s):  
Short Circuit ◽  
Short Circuit Current ◽  
Transport Processes ◽  
Rapid Decline ◽  
Tissue Viability ◽  
Bathing Medium ◽  
Flux Measurements ◽  
Unidirectional Fluxes ◽  
K Transport ◽  
Epithelial Resistance

Recta of desert locusts were mounted as flat sheets in ‘Ussing-type’ chambers and various parameters of tissue viability were monitored. The trans-epithelial resistance, the electropotential difference, the short-circuit current (Isc), and unidirectional fluxes of 22Na+, 36C1− and 42K+ all remained relatively constant during the 3rd and 4th h. The direction of the Isc indicated a net transport of either anions to the haemocoel, or cations to the lumen side. This current was abolished by KCN and was sensitive to temperature (Q10 = 2.4). There was a rapid decline in Isc over the first 2 h, which could be abolished by substituting NO3− or SO42- for Cl− in the bathing medium, indicating that this fall in current is due to a decline in the rate of Cl− transport. Measurements of 36C1− fluxes under short-circuit conditions confirm this interpretation. In the steady-state (3rd and 4th h), however, the same anion substitutions had no effect on IsC. Concurrent flux measurements indicated that net Na+ and K+ transport to the haemocoel side equals or slightly exceeds that of C1− in the same direction. Consequently all of the Isc must be due to unidentified ion transport processes. Transport of H+ to the lumen, or HCO3− and organic anions to the haemocoel side, is proposed.

scholarly journals Na+ and K+ transport at basolateral membranes of epithelial cells. II. K+ efflux and stoichiometry of the Na,K-ATPase.

1986 ◽  
Vol 87 (3) ◽  
pp. 485-502 ◽  
Author(s):  
T C Cox ◽  
S I Helman
Keyword(s):  
Short Circuit ◽  
Short Circuit Current ◽  
Pump Current ◽  
Ringer Solution ◽  
K Channel ◽  
Na Transport ◽  
Transepithelial Na Transport ◽  
Dependent Component ◽  
Na Efflux ◽  
K Transport

Changes of 42K efflux (J23K) caused by ouabain and/or furosemide were measured in isolated epithelia of frog skin. From the kinetics of 42K influx (J32K) studied first over 8-9 h, K+ appeared to be distributed into readily and poorly exchangeable cellular pools of K+. The readily exchangeable pool of K+ was increased by amiloride and decreased by ouabain and/or K+-free extracellular Ringer solution. 42K efflux studies were carried out with tissues shortcircuited in chambers. Ouabain caused an immediate (less than 1 min) increase of the 42K efflux to approximately 174% of control in tissues incubated either in SO4-Ringer solution or in Cl-Ringer solution containing furosemide. Whereas furosemide had no effect on J23K in control tissues bathed in Cl-rich or Cl-free solutions, ouabain induced a furosemide-inhibitable and time-dependent increase of a neutral Cl-dependent component of the J23K. Electroconductive K+ transport occurred via a single-filing K+ channel with an n' of 2.9 K+ efflux before ouabain, normalized to post-ouabain (+/- furosemide) values of short-circuit current, averaged 8-10 microA/cm2. In agreement with the conclusions of the preceding article, the macroscopic stoichiometry of ouabain-inhibitable Na+/K+ exchange by the pump was variable, ranging between 1.7 and 7.2. With increasing rates of transepithelial Na+ transport, pump-mediated K+ influx saturated, whereas Na+ efflux continued to increase with increases of pump current. In the usual range of transepithelial Na+ transport, regulation of Na+ transport occurs via changes of pump-mediated Na+ efflux, with no obligatory coupling to pump-mediated K+ influx.

scholarly journals Active Transport of Sodium by the Isolated Midgut of Hyalophora Cecropia

1971 ◽  
Vol 54 (1) ◽  
pp. 269-274
Author(s):  
W. R. HARVEY ◽  
K. ZERAHN
Keyword(s):  
Active Transport ◽  
Sodium Transport ◽  
Transport Mechanism ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Large Current ◽  
Sodium Flux ◽  
Hyalophora Cecropia ◽  
Sodium Solution

1. Sodium and lithium are actively transported by the isolated midgut of Hyalophora cecropia. 2. The short-circuit current in 32 mM sodium solution is about half of that in 32 mM potassium solution. 3. The sodium flux measured with 22Na from blood-side to lumen accounts for all of the short-circuit current and is 19 times the flux from lumen to blood-side. 4. In a solution containing 16 mM potassium and 16 mM sodium there is no transport of sodium, although a large current remains. 5. The sodium transport mechanism is not sensitive to ouabain.

Ion transport in isolated cornea of the rabbit

1965 ◽  
Vol 209 (6) ◽  
pp. 1311-1316 ◽  
Author(s):  
K. Green
Keyword(s):  
Ion Transport ◽  
Active Transport ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Ringer Solution ◽  
Initial Value ◽  
Rabbit Cornea ◽  
Flux Measurements ◽  
Net Flux ◽  
Normal Ringer

When the isolated rabbit cornea is bathed with well-stirred normal Ringer solution, only a low potential difference (PD) exists across the tissue; the initial value of 2 mv rises to 6 mv (endothelium positive) 1 hr after excision from the animal. In sodium-free Ringer solution the PD becomes negative before becoming negligible, while in chloride-free Ringer the PD rises to triple the value in normal Ringer. Flux measurements of sodium 22 show that there is an initial inequality between the net flux and the measured short-circuit current (SCC), the values of which become equal 1 hr after removal of the cornea from the animal Flux measurements of chloride 36 during this 1st hr indicate an active transport of chloride inward across the cornea, but after 1 hr the fluxes are equal in each direction. The differences of the net currents generated by the sodium and chloride transports equal the measured SCC, and the two transports have been shown to be able to exist independently.

THE EFFECT OF VALINOMYCIN ON THE TOAD BLADDER: ANTAGONISM TO VASOPRESSIN AND ALDOSTERONE

1969 ◽  
Vol 45 (2) ◽  
pp. 287-295 ◽  
Author(s):  
P. J. BENTLEY
Keyword(s):  
Macrolide Antibiotic ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Water Transfer ◽  
Toad Bladder ◽  
Na Transport ◽  
Bathing Medium ◽  
Sodium Transfer ◽  
Sites Of Action ◽  
Subsequent Addition

SUMMARY The macrolide antibiotic valinomycin decreased short-circuit current (SCC, Na transport) across the isolated bladder of the toad. This effect was not overcome by increasing the K+ levels in the bathing medium or by the action of amphotericin B. The effects of vasopressin on both sodium and water transfer across the toad bladder were inhibited by valinomycin and the latter inhibition is non-competitive. The action of theophylline in increasing water transfer across the bladder was also inhibited. Cyclic AMP also increased water and Na+ transfer across the bladder but its action was not reduced by the macrolide. These results suggest that valinomycin inhibits adenyl cyclase. Aldosterone increases sodium transport across the toad bladder and this action was abolished by previous incubation of the tissue with the macrolide. Once the steroid-induced effect had been established subsequent addition of valinomycin did not alter the sodium transfer. Valinomycin thus appears to have several sites of action on the toad bladder.

A primary cation transport by a V-type ATPase of low specificity

1996 ◽  
Vol 199 (6) ◽  
pp. 1327-1334 ◽  
Author(s):  
J Küppers ◽  
I Bunse
Keyword(s):  
Ion Transport ◽  
Proton Transport ◽  
Short Circuit ◽  
Membrane Conductance ◽  
Short Circuit Current ◽  
Potential Difference ◽  
Lepidopteran Larvae ◽  
The Family ◽  
K Transport

The enzyme involved in outward K+ transport in insect epithelia belongs to the family of V-ATPases. Evidence has been reported relating the generation of the K+ gradient to a primary electrogenic proton transport via a distinct electrophoretic nH+/K+ antiport. The subject of this paper is the transport of K+ at a thread hair sensillum of the cockroach in situ. We recorded changes in the voltage and resistance of the ion-transporting membrane and of shifts in pH caused by inhibition of energy metabolism and by putative inhibitors of a proton/cation exchanger. The results are supplemented by previous determinations of the K+ activities in the same preparation. 1. In cockroach hair sensilla, the ion transport generates a membrane voltage of 105 mV. We found that the transport rendered the positive output compartment alkaline with respect to the cytoplasm by 1.0 pH unit compared with the pH at equilibrium distribution, and we infer that proton transport cannot be the process that energizes the generation of the K+ gradient. 2. The ion transport created an electrochemical potential difference for protons, DeltaetaH, of approximately 4.5 kJ mol-1, while the potential difference for K+, DeltaetaK, amounted to approximately 11 kJ mol-1. Both potential differences are directed to the cytosol. It follows from DeltaetaK/DeltaetaH that an antiport would have to be electrophoretic to drive K+ by DeltaetaH and it should, therefore, contribute to the membrane conductance. Amiloride and harmaline did not significantly change the pH in the adjacent spaces and did not affect the voltage or the resistance of the transporting membrane. Previous determinations of the impedance have shown that the ATP-independent conductance of this membrane is small, supporting the conclusion that it lacks an electrophoretic antiport. From these results, we deduce that K+ transport in cockroach sensilla is not secondary to a proton transport and an electrochemical proton gradient. The phenomena observed match the performance of a primary, electrogenic, cation-translocating ATPase of the type deduced from analyses of the short-circuit current at the midgut epithelium of lepidopteran larvae. The validity of the H+ transport/antiport hypothesis is discussed.

Transepithelial transport kinetics and Na entry in frog skin: effects of novobiocin

1976 ◽  
Vol 231 (6) ◽  
pp. 1866-1874 ◽  
Author(s):  
LJ Cruz ◽  
TU Biber
Keyword(s):  
Kinetic Parameters ◽  
Frog Skin ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Close Correlation ◽  
Transport Kinetics ◽  
Transepithelial Transport ◽  
Linear Component ◽  
Na Transport ◽  
Transepithelial Na Transport

Na+ entry across the outer surface of frog skin and transepithelial Na transport were studied simultaneously at different [Na] in either the presence or absence of novobiocin by direct measurements of J12 (unidirectional uptake) and Io (short-circuit current). J12 consisted of two components: one linear, the other saturable. The kinetic parameters of the saturating components in controls were close to the kinetic parameters of overall transepithelial transport (Jm12 = 1.68+/-0.13 mleq cm-2h-1; Io =1.80+/-0.14 mueq cm-2h-1. K12 = 6.02+/-1.27 mM;Kio=6.12+/-1.33 mM). Novobiocin significantly augmented net transepithelial Na transport by increasing J13. J31 remained unaffected. A 1:1 relationship between the saturating component of J12 and Io was observed in both treated and untreated skins at all [Na] tested. (Jm12Iom, k12, and Kio were significantly larger in treated skins, but despite very drastic changes in transport rates, a close correlation between kinetic parameters of entry step and transepithelial transport was maintained. This suggests that the kinetics of transepithelial transport may simply reflect those of the rate-limiting step: the Na entry across the outer barrier of the skin. The results indicate that the linear component of J12 is not involved in transepithelial transport kinetics.

Steroid hormone stimulation of Na+ transport in A6 cells is mediated via glucocorticoid receptors

1993 ◽  
Vol 264 (4) ◽  
pp. C875-C884 ◽  
Author(s):  
T. J. Schmidt ◽  
R. F. Husted ◽  
J. B. Stokes
Keyword(s):  
Cell Line ◽  
Glucocorticoid Receptors ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Mineralocorticoid Receptors ◽  
Na Transport ◽  
Receptor Complexes ◽  
Binding Forms ◽  
Mineralocorticoid Antagonists

The A6 cell line derived from the toad kidney forms polarized, highly differentiated epithelial monolayers in culture and has been utilized as an experimental model for studying regulation of transepithelial Na+ transport by aldosterone. In the present study we evaluated the specific role(s) of glucocorticoid and mineralocorticoid receptors in mediating this enhanced electrogenic Na+ transport, which was measured experimentally as an increase in short-circuit current (Isc). Our data demonstrate that specific glucocorticoid agonists (100 nM), including RU 28362 and RU 26988, elicit “mineralocorticoid-like” increases in Isc that are blocked by the glucocorticoid antagonist RU 38486 but are unaffected by mineralocorticoid antagonists including RU 28318 and RU 26752. The stimulatory effects of aldosterone (100 nM) were also blocked by RU 38486 and not by mineralocorticoid antagonists. These data extend earlier studies suggesting that in this cell line aldosterone mediates its physiological effects via binding with relatively low affinity (dissociation constant Kd congruent to 25-50 nM) to glucocorticoid receptors, despite the presence of apparently normal mineralocorticoid receptors. Our in vitro biochemical studies also demonstrate that A6 glucocorticoid receptor complexes can be thermally activated or transformed to DNA binding forms which exhibitaltered elution profiles from anion-exchange resins. Thus, based on several criteria, these amphibian glucocorticoid receptors appear very similar to classical mammalian receptors and are capable of mediating all of the stimulatory effects of aldosterone on net Na+ transport.

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