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C C Aickin

Publications and source records attributed to C C Aickin.

16 recordsLinked to original sources
PubMed1990

Effect of Na+ and K+ on Cl- distribution in guinea-pig vas deferens smooth muscle: evidence for Na+, K+, Cl- co-transport.

1. Smooth muscle cells of the guinea-pig vas deferens after Cl- depletion actively reaccumulate ions to a level many times higher than that predicted by a passive distribution, even when anion exchange (largely responsible for Cl- movements in this preparation) is inhibited by DIDS (4,4'-diisothiocyanostilbene-2,2'-disulphonic acid). The cells therefore must possess a second mechanism for Cl- accumulation. We have now investigated the ionic requirement of this mechanism using a combination of ion analysis, 36Cl fluxes and direct measurement of the intracellular Cl- activity (aiCl). 2. In the steady state, the Cl- content of tissues was 12-16% less in Na(+)-free solution than in normal Krebs solution. 3. Loss of 36Cl into Cl(-)-free solution was slowed by the absence of Na+ and accelerated on its readdition. Uptake of 36Cl by Cl(-)-depleted tissues was also reduced in the absence of extracellular Na+, particularly at longer time intervals as uptake reached completion. These effects occurred in the presence and absence of CO2-HCO3- and in the presence of DIDS. 4. The initial rate of rise of aiCl on readdition of Cl- to Cl(-)-depleted cells was not significantly affected by the absence of Na+ in the presence of a functional anion exchange, but aiCl stabilized at a lower value than in normal solution. Readdition of Na+ stimulated a rise in aiCl to the control level. Removal and readdition of K+ under these conditions had negligible effects. 5. When anion exchange was inhibited by the presence of DIDS, removal and readdition of Na+ caused, respectively, a marked inhibition and stimulation of the rise in aiCl during Cl- reaccumulation. Under these conditions removal and readdition of K+ had similar effects. 6. The results suggest that Na+, K+, Cl- co-transport is involved in transmembrane movements of Cl- at least when the anion exchange mechanism is blocked. 7. The possibility that the marked effects of changes in external Na+ on transmembrane Cl- movements in the presence of a functional anion exchange mechanism are caused by secondary effects due to changes in intracellular pH as well as to suppression of Na+, K+, Cl- co-transport is discussed.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed1990

The effect of loop diuretics on Cl- transport in smooth muscle of the guinea-pig vas deferens and taenia from the caecum.

1. The role of Na+, K+, Cl- co-transport, identified in the previous paper (Aickin & Brading, 1990), has been characterized further by investigation of the effects of loop diuretics on Cl- movements in the smooth muscle cells of guinea-pig vas deferens measured by 36Cl fluxes and Cl(-)-sensitive microelectrodes. Some flux experiments were also repeated in the taenia from the guinea-pig caecum. 2. Frusemide (2 mM) reduced the steady-state Cl- content, slowed 36Cl loss into Cl(-)-free solution and both slowed and reduced Cl- accumulation by Cl(-)-depleted cells of the vas deferens. When anion exchange was inhibited by the presence of DIDS, (4,4'-diisothiocyanostilbene-2,2'-disulphonic acid), frusemide further slowed the loss of Cl- into Cl(-)-free solution, further reduced Cl- accumulation such that Cl- uptake amounted to a level consistent with a passive distribution and halted the rise in the intracellular Cl- activity (aiCl) at levels above about 10 mM. 3. Application of the higher-affinity loop diuretics bumetanide and piretanide in vas deferns had no significant effect on 36Cl efflux into Cl(-)-free solution or on the initial rate of rise of aiCl but reduced the final level attained. In the presence of DIDS, however, both agents further slowed efflux into Cl(-)-free solution, and halted the rise in aiCl at levels above about 10 mM. Measurement of greatly slowed intracellular pH transients on removal and readdition of external Cl- (Clo-) in the presence of frusemide suggests that the larger effects of this drug are mediated by inhibition of anion exchange as well as of co-transport. 4. The relative potency of the loop diuretics, investigated in the presence of DIDS was: bumetanide greater than piretanide greater than frusemide. This sequence was found in both vas deferens, using direct measurement of aiCl, and taenia, using 36Cl uptake. 5. Comparison of data from the vas and taenia showed that 36Cl efflux into Cl(-)-free, HCO3(-)-free solution was about twice as fast in the taenia, and that bumetanide or piretanide reduced this efflux to about the same rate as that observed in the vas with or without the loop diuretic. DIDS caused a similar absolute reduction of efflux in both preparations. 6. Stimulation of 36Cl efflux on readdition, and inhibition on removal of Nao+ in the presence of DIDS, was much greater in the taenia than in vas and in both preparations was blocked by bumetanide or piretanide.(ABSTRACT TRUNCATED AT 400 WORDS)

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed1989

Intracellular chloride and the mechanism for its accumulation in rat lumbrical muscle.

1. Double-barrelled Cl(-)-sensitive microelectrodes have been used to measure the intracellular Cl- activity (aCli) and membrane potential (Em) in rat lumbrical muscles. The mean Cl- equilibrium potential (ECl), calculated from the measured aCli in sixty fibres, was 2.9 +/- 2.5 mV (S.D. of an observation) less negative than Em. The value of aCli was higher than would be expected for a passive distribution, by a mean 1.4 +/- 1.2 mM. The mean Em was -59.5 +/- 8.2 mV. 2. Removal of external Cl- (Cl-(o)) resulted in a rapid fall in aCli and a transient depolarization. aCli stabilized at an apparent level of 1.7 +/- 1.0 mM (n = 24) while Em became substantially more negative than in normal Krebs solution (mean, -80.1 +/- 12.4 mV). Readdition of Cl-(o) caused a rapid rise in aCli and transient hyperpolarization. ECl quickly became less negative than Em and both then fell in parallel towards the levels previously recorded in normal Krebs solution. 3. If lack of selectivity of the Cl(-)-sensitive ion exchanger and the intracellular presence of interfering anions, assumed to be responsible for the apparent aCli recorded in Cl(-)-depleted fibres, were also responsible for the apparently non-passive Cl- distribution recorded under normal conditions, the difference between the calculated ECl and Em would increase at more negative potentials. This was not observed over a range of Em values between -46 and -84 mV. 4. Inhibition of the Cl- permeability by application of 9-anthracene carboxylic acid (9-AC) resulted in an immediate rise in aCli and hyperpolarization. An aCli up to 40 mM higher, or eleven times higher, than that predicted by a passive distribution was recorded. Application of 9-AC after depletion of intracellular Cl- in Cl(-)-free solution had no effect on either the apparent aCli or Em. 5. It is concluded that Cl- ions are actively accumulated by the skeletal muscle fibre and that the Cl- distribution therefore normally exerts a depolarizing influence. 6. In the presence of 9-AC and nominal absence of CO2 and HCO3-, readdition of Cl-(o) to Cl(-)-depleted fibres resulted in a substantial rise in aCli and a small, maintained depolarization. This clear demonstration of active accumulation was used to investigate the mechanism responsible for inward transport of Cl- ions. 7. Neither application of CO2 and HCO3- nor application of DIDS (4,4'-diisothiocyanostilbene-2,2'-disulphonic acid) had any effect on the accumulation of Cl- ions. This suggests that Cl(-)-HCO3- exchange is not involved.(ABSTRACT TRUNCATED AT 400 WORDS)

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed1988

Movement of acid equivalents across the mammalian smooth muscle cell membrane.

Factors affecting intracellular pH (pHi) in the smooth muscle of guinea pig ureter have been investigated using pH-sensitive microelectrodes. Associated acids and bases appear to have free passage across the cell membrane but results suggest very low permeability to charged acid equivalents, thus implicating carrier-mediated movements in many of the observed pHi transients. Recovery from acidosis in the nominal absence of CO2 was inhibited by removal of Na+ and by the presence of amiloride, indicating that Na+/H+ exchange was responsible. The presence of CO2 resulted in a faster recovery from acidosis but, since intracellular buffering power was not increased, not a substantially faster effective extrusion of protons. Surprisingly, amiloride no longer caused discernable inhibition. Recovery from moderate acidosis remained Na+ dependent but was not inhibited by DIDS or acetazolamide or by the absence of Cl-, suggesting a dominant Na+-, and HCO3(-)-dependent mechanism unlike any hitherto described. Recovery from alkalosis was inhibited by DIDS and Cl(-)-free conditions, indicating that Cl-/HCO3- exchange was involved. Results suggest reversal of this mechanism on extreme acidosis. Experiments in vascular smooth muscle with fluorescent indicators confirm the presence of Na+/H+ exchange but provide conflicting evidence about the presence and properties of the HCO3(-)-dependent mechanism.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed1987

Investigation of factors affecting the intracellular sodium activity in the smooth muscle of guinea-pig ureter.

1. The intracellular Na+ activity (aNai) of the smooth muscle cells from guinea-pig ureter has been measured using double-barrelled Na+-sensitive micro-electrodes. aiNa in modified Krebs solution at 35 degrees C was of a mean 7.4 +/- 2.9 mM (n = 32, S.D. of an observation), equivalent to a Na+ equilibrium potential (ENa) of +66.7 mV. Membrane potential (Em) was of a mean -50.8 +/- 4.6 mV. 2. Inhibition of the Na+ pump by application of ouabain or removal of external K+ (K+o) resulted in a restricted rise of aNai. The rate of rise was faster in the presence of ouabain (10(-4) M) but the stabilized aNai was not significantly different from that observed after the prolonged absence of K+o. The mean aiNa recorded after prolonged Na+ pump inhibition was 20.6 +/- 5.5 mM (n = 28), equivalent to an ENa of +39.6 mV. Neither removal of K+o after aNai had stabilized in the presence of ouabain nor application of ouabain after aNai had stabilized in K+-free solution caused a rise in aiNa, suggesting that the Na+ pump was fully inhibited by either procedure. 3. Reduction of Na+o resulted in a rapid fall in aiNa against the electrochemical gradient, both before and after Na+ pump inhibition. At each level of Na+o, aNai stabilized such that ENa remained approximately constant in either condition. Readdition of Na+o resulted in a rapid recovery of aNai. 4. Elevation of Ca2+o (at constant Na+o) caused a fall in aNai of much the same time course as that observed on reduction of Na+o, both before and after Na+ pump inhibition. The extent of the fall was dependent upon the initial aNai. Reduction of Ca2+o resulted in a rise in aNai. 5. Elevation of the external divalent cation concentration with Mn2+ or, to a lesser extent, Mg2+ reduced aiNa in the presence of a functional Na+ pump. But after prolonged exposure to ouabain or K+-free solution, elevation of Mg2+o had no effect on aiNa while application of Mn2+o caused a slow rise. These results suggest that Ca2+o affects aiNa in two ways. One is mimicked by Mg2+ and Mn2+ and is probably due to alteration of the Na+ leak. The other is a specific effect, revealed by Na+ pump inhibition. 6. It is concluded that aiNa can be maintained far from equilibrium in the absence of a functional Na+ pump. Several lines of evidence are discussed which indicate the participation of Na+-Ca2+ exchange in Na+ extrusion in this condition.

Action Potentials↗
PubMed1987

An investigation of sodium-calcium exchange in the smooth muscle of guinea-pig ureter.

1. After application of ouabain (10(-4) M), the intracellular Na+ activity (alpha iNa) of smooth muscle cells in the guinea-pig ureter stabilizes at a relatively low level which can be rapidly lowered by reduction of external Na+ (Na+o) or elevation of Ca2+o. Both these procedures also elicit a transient contracture. These observations have previously been interpreted as evidence for Na+-Ca2+ exchange. The presence of such an exchange mechanism has now been further investigated by measurements of alpha iNa, tension, ion analysis and 22Na efflux. 2. Ion analysis demonstrated that tissues were able to maintain a high cellular K+ content in the presence of ouabain, but slowly lost K+ and gained Na+ if K+o was also removed, as expected for an infinite outward gradient for K+ and a fully inhibited Na+ pump. 3. Tissues were only able to maintain a low cellular Na+ and high cellular K+ in the presence of ouabain if Ca2+ was present in the bathing solution. Reduction of Ca2+o to very low levels also caused a continual slow rise in alpha iNa in the presence of ouabain, provided that the prolonged depolarization caused by these low levels was prevented by elevation of Mg2+o. Alteration of the membrane potential by changing K+o at constant Na+o showed that alpha iNa decreased by about 1.2 mM for a 10 mV depolarization, within the range from -70 to -30 mV. 4. A small Ca2+o-activated 22Na efflux was observed in ouabain-treated tissues in the absence of Na+o. 40 mM-Ca2+ was not more effective at activating this efflux than was 2.5 mM-Ca2+, while 40 mM-Mg2+ was ineffective. Restoration of the normal Na+o caused a large increase in the rate of 22Na loss. 5. Application of Mn2+ in the presence of ouabain caused a slow rise in alpha iNa and a small decline in resting tension. The fall in alpha iNa on reduction of Na+o was slowed by the presence of Mn2+ (mean half-time increased from 1.7 to 5.0 min) and the concomitant contracture was almost abolished. These results are consistent with a Mn2+-induced inhibition of Na+-Ca2+ exchange. However, the fall in alpha iNa induced by elevation of Ca2+o was unaffected by the presence of Mn2+ and the attendant contracture was, if anything, enhanced. 6. Observation of changes in alpha iNa and tension at various Mn2+ and Ca2+ concentrations demonstrated a competitive interaction between the two divalent cations.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗
PubMed1986

Intracellular pH regulation by vertebrate muscle.

Regulation of pHi in the face of acidosis resulting from contracture would appear to be of such fundamental importance to the physiology of the muscle cell that a process common to all muscle types seems a reasonable prediction. However, this has not been found to be the case. The transmembrane Na+ gradient clearly plays a major role and the process appears to be electroneutral in all three classes of muscle, but the transport mechanisms, even within the mammal, are different. It is an interesting observation that the ability of the muscle cell to regulate pHi in the presence of CO2, presumably governed by PHCO3, is related to PCl although there is little evidence for HCO3- permeation through Cl- channels. Virtually no recovery from CO2-induced acidosis is observed in normally polarized frog skeletal muscle, where PCl forms a large part of the resting conductance, whereas the same steady state pHi is recorded in the presence of various CO2 levels in mammalian smooth muscle, where PCl is very low. The study of pHi regulation in vertebrate muscle has provided important lessons for the subject as a whole. Experience in cardiac muscle has shown that if Na+-Ca2+ exchange is present, great care is required in interpretation of results where the transmembrane Na+ gradient is altered or where Ca2+ levels are changed. Interpretation may be even more complex, bearing in mind the recent reports that Ca2+ inhibits Na+-H+ exchange. "Indeed," it seems appropriate to conclude, "if a little knowledge is dangerous, where is the man who has so much as to be out of danger?" (Thomas Huxley).

Acidosis↗
PubMed1985

The effects of bicarbonate and foreign anions on chloride transport in smooth muscle of the guinea-pig vas deferens.

The selectivity of the external site of the Cl transporting mechanism in the guinea-pig vas deferens has been investigated by measurement of 36Cl uptake and efflux and by direct measurement of intracellular pH. Replacing 50% of the Cl in normal Krebs solution inhibited the 15 min uptake of 36Cl in the order NO3 greater than Br greater than SCN greater than F greater than I greater than glucuronate, both in Cl-depleted tissues and tissues pre-incubated in the 50%-Cl solutions (steady-state uptake). After 3 h incubation in these solutions, the total cellular Cl was reduced by the anions in the order Br greater than NO3 greater than I greater than SCN greater than F greater than glucuronate. Br, NO3 and I reduced the cellular Cl to less than 50% of normal, suggesting that they are actively taken up by the cells. The ability of foreign anions to inhibit a 3 min uptake of high specific activity, low concentration Cl (6.5 mM) suggests an apparent affinity series of NO3 greater than Cl = SCN = Br greater than I greater than F at the external site. Addition of NO3, Cl, Br, HCO3, F, SCN or I to a Cl-free, nominally HCO3-free bathing solution accelerated 36Cl efflux. The first four mentioned were powerful stimulants, the other three less potent. However, the exact position of HCO3 in the sequence is uncertain. The rapidity with which CO2 crosses the membrane and forms HCO3 intracellularly may allow competition between HCO3 and Cl at the internal site and so distort the result. The action of F is also questionable since this ion drastically reduces the divalent cation activity and is a metabolic inhibitor. Measurement of intracellular pH provided conclusive evidence that Cl, NO3, Br and I can exchange with HCO3 across the cell membrane. This exchange is as rapid with NO3 as with Cl but slower with Br and considerably slower with I. The results also indicate that SCN ions cross the cell membrane. It is concluded that Cl, HCO3, Br and NO3 are all translocated by the exchange carrier. I and perhaps SCN also interact with the transport mechanism, but the translocation rate is then greatly reduced. The precise order of the affinity of these anions remains uncertain but the following sequence: NO3 greater than Cl = SCN = Br greater than I greater than F is considered to be the most likely.

Animals↗
PubMed1979

Decrease of inhibitory driving force in crayfish stretch reception: a mechanism of the convulsant action of penicillin.

The effect of penicillin on the evoked IPSP was investigated in the isolated crayfish stretch receptor. The IPSP driving force (IPSP reversal potential minus membrane potential) was reduced in a dose-dependent fashion but, when necessary correction was made for the decrease in resting membrane conductance, the synaptic conductance was only slightly reduced. The possibility that a penicillin-induced intracellular acidification was responsible for the decrease in IPSP driving force is considered.

Animals↗
PubMed1977

An investigation of the ionic mechanism of intracellular pH regulation in mouse soleus muscle fibres.

1. Intracellular pH (pH(i)) of surface fibres of the mouse soleus muscle was measured in vitro by recessed-tip pH-sensitive micro-electrodes. pH(i) was displaced in an acid direction by removal of external (NH(4))(2)SO(4) after a short exposure, and the mechanism of recovery from this acidification was investigated.2. Removal of external K caused a very slow acidification (probably due to the decreasing Na gradient) but had no effect on the rate of pH(i) recovery following acidification. This indicates that K(+)-H(+) exchange is not involved in the pH(i) regulating system.3. Short applications of 10(-4)M ouabain had no obvious effect on pH(i) and did not alter the rate of pH(i) recovery following acidification. This suggests that there is no direct connexion between the regulation of pH(i) and the Na pump.4. Reduction of external Ca from 10 to 1 mM caused a transient fall in pH(i), but the rate of pH(i) recovery following acidification was unaffected. This suggests that Ca(2+)-H(+) exchange is not involved in the pH(i) regulating system.5. An 11% reduction in external Na caused a significant slowing of pH(i) recovery following acidification. 90% or complete removal of external Na almost stopped pH(i) recovery. This suggests that Na(+)-H(+) exchange is involved in pH(i) regulation.6. Amiloride (10(-4)M) reversibly reduced the rate of pH(i) recovery to much the same extent as removal of external Na. Its effect was not additive to that of removal of external Na.7. Internal Na ion concentration ([Na(+)](i)), measured using Na(+)-sensitive micro-electrodes, fell on application of (NH(4))(2)SO(4) and increased on its removal. The increase transiently raised [Na(+)](i) above the level recorded before (NH(4))(2)SO(4) application. This overshoot of [Na(+)](i) was almost completely inhibited by amiloride. This is consistent with the involvement of Na(+)-H(+) exchange in the pH(i) regulating system.8. Removal of external CO(2) or application of SITS (10(-4)M) caused some slowing of the rate of pH(i) recovery following acidification by removal of (NH(4))(2)SO(4). The effect of SITS was additive to that of Na-free Ringer or amiloride. These results suggest that Cl(-)-HCO(3) (-) exchange is also involved in the pH(i) regulating system and that it is a separate mechanism. Under the conditions used, Cl(-)-HCO(3) (-) exchange formed about 20% of the pH(i) regulating system.9. Decreasing the temperature from 37 to 28 degrees C not only caused an increase in pH(i), but also considerably slowed the rate of pH(i) recovery following acidification. We have calculated a Q(10) for Na(+)-H(+) exchange of 1.4 and for Cl(-)-HCO(3) (-) exchange, 6.9.10. We conclude that the pH(i) regulating system is comprised of two separate ionic exchange mechanisms. The major mechanism is Na(+)-H(+) exchange, which is probably driven by the transmembrane Na gradient. The other mechanism is Cl(-)-HCO(3) (-) exchange, which probably requires metabolic energy.

Amiloride↗
PubMed1975

Micro-electrode measurement of the internal pH of crab muscle fibres.

The internal pH of crab muscle fibres was measured using recessed-tip pH-sensitive micro-electrodes. Immediately following electrode penetration the mean internal pH was 7-21 +/- 0-02 (S.E. of mean) and the mean membrane potential was -64-9 +/- 0-6 mV (S.E. of mean). If H+ ions were passively distributed across the fibre membrane the internal pH would have been 6-39. 2. The internal pH tended to rise before stabilizing at a mean value of 7-27 +/- 0-02 (S.E. of mean). The difference between immediate and stabilized values is highly significant and suggests acid injury on electrode penetration. 3. Changing the membrane potential or external pH had only small, slow effects on internal pH. 4. External CO2 caused a large and rapid decrease in internal pH. With low concentrations of CO2, the effect was dependent on the initial pH as predicted by the Law of Mass Action. During a long exposure to 2-65% CO2 at pH 7-5, the internal pH returned slowly to its previous value, suggesting active transport of H+ (or OH- or HCO3-) ions across the fibre membrane. 5. The internal buffering power calculated from the response to 2-65% CO2 was 47-3 +/- 2-8 slykes (m-equiv H+/pH unit per l.) (S.E. of mean).

Animals↗