SEARCH · PubMed Health

Results for “Isethionic Acid”

Explore indexed PubMed citations for clinical trials, systematic reviews and public health research. Read source abstracts and follow each citation to its original PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 235 records · Page 13Linked to original sources

PubMed1996

Parapodial swim muscle in Aplysia brasiliana. II. Ca(2+)-dependent K+ currents in isolated muscle fibers and their blockade by chloride substitutes.

1. We describe here the properties of two Ca(2+)-dependent K+ currents found in type II muscle fibers dissociated from the parapodia (swim appendages) of the marine snail Aplysia brasiliana. 2. Type II parapodial muscle fibers display three voltage-dependent currents that are also seen in type I fibers, a delayed rectifier current [IK(V)], a transient A current (IA), and a prominent L-type Ca2+ current. In addition, type II fibers also have two outward K+ currents, a transient, inactivating one and a slower, noninactivating one [IK(Ca,t) and IK(Ca,s), respectively], that are Ca2+ dependent. The expression of these currents in normal type II fibers generally produces a waveform of total outward current that is faster to peak than the total outward current seen in response to voltage steps in type I fibers and that does not inactivate at the end of an 80-ms voltage step. 3. Both IK(Ca,t) and IK(Ca,s) are absent when external Ca2+ is eliminated or when extracellular Ca2+ concentration ([Ca2+]o) is substituted with 10 mM Co2+ or Ba2+. Their threshold for activation is around -40 mV. IK(Ca,t) peaks rapidly and then inactivates, but IK(Ca,s) rises slowly and does not inactivate for as long as 200 ms. Both currents, like IK(V) and IA, are sensitive to tetraethylammonium and 4-aminopyridine and are not readily separated from either the voltage-gated currents or from one another by these pharmacological agents. 4. Tail current analysis from depolarized voltage steps in varying (K+]o demonstrates that these currents are carried by K+ ions and not by Cl-. 5. An unexpected finding, however, is that these Ca(2+)-dependent K+ currents are blocked by standard Cl- ion substitutes, such as methanesulfonate, isethionate, and propionate. IK(Ca,s) is slightly more sensitive to these Cl- substitutes than is IK(Ca,t). The chloride blocker 4,4'-diisothiocyantastilbene-2,2'disulfonic acid also partially blocked the Ca(2+)-dependent K+ currents. 6. The presence of these Ca(2+)-dependent K+ currents in type II fibers may contribute to a more rapid repolarization following depolarization-induced contractions. In contrast to type I fibers, which have smaller calcium current and no Ca(2+)-activated K+ currents, type II muscle cells may function more like "fast" fibers and relax more rapidly.

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

PubMed1992

Purification and characterization of proteinase In, a trypsin-like proteinase, in Escherichia coli.

We previously found a trypsin-like proteinase which momentarily appears immediately before DNA synthesis in the cell cycle of Escherichia coli synchronized by phosphate starvation and which is closely related to the initiation of DNA replication (Kato, M., Irisawa, T., Morimoto, Y. and Muramatu, M., unpublished results). The proteinase was named proteinase In. It was purified approximately 2880-fold with a recovery of 15%. The isolated enzyme appeared homogeneous by gel filtration and electrophoresis. Its molecular mass was estimated by analytical gel filtration and SDS/PAGE as approximately 66 kDa. The isoelectric point of proteinase In is 4.9 and its optimal pH is approximately 9. Although protein In hydrolyzes fluorogenic substrate for trypsin, its hydrolytic activity seems markedly affected by amino-acid sequence lying towards the N-terminal from the P1 (lysine, arginine) residue. The proteinase does not hydrolyze N2-benzoyl-D,L-arginine-4-nitronanilide and fluorogenic substrates for chymotrypsin and elastase. The proteinase activity is inhibited by leupeptin, antipain and 4-nitrophenyl 4-guanidinobenzoate, but the effects of tosyl-L-lysine chloromethane, diisopropylfluorophosphate, benzamidine and pentamidine isethionate on the proteinase activity are weak or not inhibitory. Its activity is strongly affected in the presence of NaCl and KCl, and at a concentration of 1.5 M, these increase the activity 14-fold and 13-fold, respectively, above that without salt. Proteinase In was strongly inhibited by various esters of trans-4-guanidinomethylcyclohexanecarboxylic acid, and their inhibitory effects were roughly correlated with those on growth of E. coli. Proteinase activity was found in the cytoplasmic fraction.

Amino Acid Sequence↗

PubMedDate not supplied

Importance of solvent drag and diffusion in bile acid-dependent bile formation: ion substitution studies in isolated perfused rat liver.

Ion substitution studies were carried out in the isolated perfused rat liver to define the importance of solvent drag and diffusion in bile acid-dependent bile formation. Two different methods, namely single injection (20 mu moles) and continuous infusions at 0.4, 0.8, 1.2, and 1.6 mu moles per min taurocholate (TC), were used to determine the bile acid-dependent bile flow (BADF). Both methods gave essentially the same results. Replacement of Na+ (146mM) by 120 or 146 mM Li+ and Cl-(127mM) by 120 mM NO3- increased BADF significantly. On the other hand, replacement of Na+ by 120 mM choline and Cl- y 120 mM isethionate decreased the BADF. The osmolarity of TC solution was not different when Na+ was replaced by 120 mM Li+ or choline and TC did not affect the osmotic activity of NaCl, and choline-Cl differently. Thus, the observed effect of Na+ replacement on BADF is not due to any change in the osmotic activity of the secreted TC. Substitution of HCO3- by equimolar tricine also decreased BADF. Under this condition, BADF increased when NaCl was replaced by equimolar NaNO3. Thus, HCO3- does not seem to be essential for TC choleresis. Since Li+ and NO3- are more permeable, and choline and isethionate are less permeable than Na+ and Cl-, respectively, these results suggest that the BADF is dependent on the permeability of the substituting cations and anions and thus support the hypothesis that solvent drag and diffusion play an important role in BADF.

Animals↗

PubMed1983

Interactions of C12 surfactants with the skin: studies on enzyme release and percutaneous absorption in vitro.

Using an in vitro penetration cell, it has been shown that enzymes (acid phosphatase, lactate dehydrogenase and N-acetylglucosaminidase) are released from rat-skin slices in response to contact with two irritant C12 surfactants, sodium laurate and sodium lauryl sulphate, but not with the non-irritant sodium lauroyl isethionate. About 3-5 hr contact of the stratum corneum with surfactant and a long incubation time (24 hr) were required for enzyme release. Adsorption and penetration of the two effective surfactants was also studied and the results for sodium lauryl sulphate suggested a relationship between enzyme release and adsorption of surfactant. However, no such simple relationship was observed for sodium laurate, emphasizing the complex nature of surfactant interactions with the skin.

Adsorption↗

PubMed1985

Intracellular pH regulation in the sensory neurone of the stretch receptor of the crayfish (Astacus fluviatilis).

The ionic mechanisms of intracellular pH (pHi) regulation were studied in the slowly adapting sensory cell of the crayfish stretch receptor by using pH-, Na+- and Cl(-)-sensitive liquid ion exchanger electrodes. Under control conditions a mean pHi of 7.23 +/- 0.12 (S.D.) at a mean membrane potential of 68.3 +/- 4.1 mV S.D. was found in sixteen cells. Thus pHi is about 1 pH unit more alkaline than predicted from passive distribution, implying the presence of an acid extrusion mechanism. In order to acidify the cytoplasm, the cell was either acid-loaded by NH4Cl or exposed to CO2 and CO2/HCO3- solutions. During CO2 exposures pHi was regulated only if calculated amounts of HCO3- were added to keep external pH (pHo) constant. The pHo per se was found to be an important determinant of pHi and its regulation. Substitution of external Na+ by choline inhibited pHi recovery almost completely. As soon as Na+ was readmitted H+ extrusion occurred immediately at a rate similar to that of the control. The internal Na+ activity (aiNa) ranged between 6 and 13 mM with a mean of approximately 9.1 +/- 2.5 mM (S.D.; n = 8). The effects of various solutions on aiNa and the temporal relationship between aiNa and pHi in NH4Cl acid-loaded cells were investigated. The amount of aiNa increased during cell internal acidification and recovered in parallel with pHi recovery in NH4Cl acid-loaded cells. Experiments with 10(-4) M-ouabain and K+-free conditions suggest that neither the Na+-K+ pump nor external K+ are directly involved in pHi regulation. The internal chloride activity (aiCl), which was lower than predicted from a passive distribution, fell during exposure to HCO3-/CO2. Regulation of pHi was inhibited if the cell was completely depleted of Cl- by prolonged exposures to Cl(-)-free solution (isethionate and/or gluconate substituted). The pHi-regulating system of the sensory cell requires Na+ and Cl- which probably operate in a combined mechanism such as Na+ -H+-Cl(-)-HCO3- or an equivalent.

Acid-Base Equilibrium↗

PubMed1989

Ascorbic acid transport in mouse and rat astrocytes is reversibly inhibited by furosemide, SITS, and DIDS.

The uptake of L-ascorbic acid (vitamin C) by astrocytes was studied using primary cultures prepared from the neopallium of newborn Swiss CD-1 mice or Sprague-Dawley rats. Initial uptake rates were significantly greater in mouse than in rat astrocytes. Exposure of cultures to 0.25 mM dibutyryl cyclic AMP for 2 weeks changed cell morphology from polygonal to stellate and stimulated ascorbate uptake, with the greatest stimulation occurring in mouse astrocytes. Uptake was specific for the vitamin since it was not diminished by the presence of other organic anions including acetate, formate, lactate, malonate, oxalate, p-aminohippurate, pyruvate and succinate. Ascorbate uptake was Na(+)-dependent but did not have a specific requirement for external Cl- (Cl-0). Substitution of Cl-0 by Br- or NO3- decreased ascorbate uptake rates by 20-31%; whereas substitution by gluconate or isethionate increased uptake by 20-31%. Ascorbate transport by astroglial cultures from both animal species was rapidly (less than or equal to 1 min) and reversibly inhibited by the anion transport inhibitors furosemide, 4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid (SITS) and 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS). The rapid and reversible effects of the impermeant inhibitors (SITS and DIDS) are consistent with direct inhibition of ascorbate transporters located in the astroglial plasma membrane.

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

PubMed1987

Efflux of putative transmitters from superfused rat brain slices induced by low chloride ion concentrations.

Slices of rat cerebral cortex, preloaded with [14C]gamma-aminobutyric acid (GABA) and either [3H]5-hydroxytryptamine (5-HT) or [3H]noradrenaline, were superfused with media in which varying concentrations of Cl- had been replaced with other monovalent anions. Rapid reduction of [Cl-], by superfusion with media containing instead the impermeant anions propionate, isethionate, gluconate, or methyl sulphate, caused increases in the efflux of tritiated biogenic amines, but the increase in that of [14C]-GABA was not significant. The increased efflux of [3H]5-HT evoked by superfusion with low Cl- levels when propionate was the replacement anion, was transient and was linearly related to the log[Cl-]-1. It was not affected by removal of Ca2+ or by addition of 10 mM Mg2+ and was delayed but not abolished by tetrodotoxin. The low Cl(-)-evoked efflux of [3H]5-HT was not affected by pretreatment with neuronal reuptake blockers but was inhibited by picrotoxin, strychnine, and 4-acetamido-4-isothiocyanostilbene-2,2-disulphonic acid and was enhanced by glycine. Muscimol and GABA were without effect. These observations are taken to indicate that the efflux of biogenic amines is brought about by terminal depolarisation due to outward movement of Cl- in low chloride-containing media. They are of relevance to other physiological and pharmacological studies in which anion concentrations are manipulated and suggest that the anion-evoked release phenomenon may provide a model for the analysis of Cl(-)-dependent mechanisms in nerve terminals.

4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfo↗

PubMed2000

Characterisation of methicillin-resistant Staphylococcus aureus from Kuwait hospitals with high-level fusidic acid resistance.

Forty-seven fusidic acid- and methicillin-resistant Staphylococcus aureus isolates from clinical samples in four hospitals in Kuwait were studied for their relatedness by biotyping and pulsed-field gel electrophoresis (PFGE) and for the genetic location of their resistance determinants. Forty-four isolates were resistant to gentamicin, kanamycin and neomycin. Forty-one isolates were resistant to erythromycin and trimethoprim, 10 were resistant to chloramphenicol and four were resistant to ciprofloxacin. They contained two or three plasmids of c. 28, 2.8 and 1.8 kb. Genetic studies demonstrated that resistance to cadmium, propamidine isethionate and ethidium bromide were linked and were carried on the c. 28-kb plasmid. Chloramphenicol resistance was encoded by the 2.8-kb plasmid in resistant isolates. No resistance was associated with the 1.8-kb plasmid and this was considered to be a cryptic plasmid. Resistance to fusidic acid, methicillin, benzylpenicillin, gentamicin, kanamycin, neomycin, tetracycline, trimethoprim, erythromycin and ciprofloxacin were located on the chromosome. All the isolates produced urease, but varied in the production of haemolysins, pigments, lipase and lecithinase and were classified into nine biotypes. In contrast, PFGE divided the isolates into two major patterns with one PFGE type constituting the majority of isolates in all four hospitals. The presence of the dominant PFGE pattern in all four hospitals suggests that it is an epidemic MRSA clone with the capacity to spread. Infection control measures should be directed towards restricting the further spread of this clone.

Anti-Bacterial Agents↗

PubMed1983

Role of inorganic electrolytes in bile acid-independent canalicular bile formation.

Ion-replacement studies were carried out in the isolated perfused rat liver to obtain insight into the role played by inorganic electrolytes in bile acid-independent canalicular bile flow (BAICF). The BAICF decreased significantly when Na+ (146 mM) was replaced by 120 mM K+, Rb+, Cs+, or choline and when Cl- (127 mM) was replaced by 120 mM acetate or isethionate; there was no reduction in BAICF when Na+ was replaced by Li+ (146 mM) and Cl- by NO-3. K+, Rb+, and Cs+, however, also caused a simultaneous decline in the perfusion rate. The BAICF decreased by 50% when HCO-3 was replaced by equimolar tricine; under this condition replacement of Cl- by NO-3, but not Na+ by Li+, decreased BAICF by 45%. Thus the hepatic transport of Cl- cannot be explained by simple diffusion only, and a special mechanism, probably Na+-coupled Cl- transport, may contribute about 30% of the BAICF. With Li+ replacing Na+ in the medium, the intracellular concentration of Li+ in isolated rat hepatocytes was less than that calculated for electrochemical equilibrium and was increased by 2 mM KCN, indicating active extrusion of this ion. Li+ was unable to activate Mg2+-ATPase of isolated rat liver plasma membranes, and 1 mM ouabain did not affect the Li+ distribution. These results suggest the potential importance of ion pumps other than Na+-K+-ATPase in BAICF.

Animals↗

PubMed1993

Spontaneous and evoked release of [3H]taurine from a P2 subcellular fraction of the rat retina.

The effects of spontaneous and evoked [3H]taurine release from a P2 fraction prepared from rat retinas were studied. The P2 fraction was preloaded with [3H]taurine under conditions of high-affinity uptake and then examined for [3H]taurine efflux utilizing superfusion techniques. Exposure of the P2 fraction to high K+ (56 mM) evoked a Ca(2+)-independent release of [3H]taurine. Li+ (56 mM) and veratridine (100 microM) had significantly less effect (8-15% and 15-30%, respectively) on releasing [3H]taurine compared to the K(+)-evoked release. 4-Aminopyridine (1 mM) had no effect on the release of [3H]taurine. The spontaneous release of [3H]taurine was also Ca(2+)-independent. When Na+ was omitted from the incubation medium K(+)-evoked [3H]taurine release was inhibited by approximately 40% at the first 5 minute depolarization period but was not affected at a second subsequent 5 minute depolarization period. The spontaneous release of [3H]taurine was inhibited by 60% in the absence of Na+. Substitution of Br- for Cl- had no effect on the release of either spontaneous or K(+)-evoked [3H]taurine release. However, substitution of the Cl- with acetate, isethionate, or gluconate decreased K(+)-evoked [3H]taurine release. Addition of taurine to the superfusion medium (homoexchange) resulted in no significant increase in [3H]taurine efflux. The taurine-transport inhibitor guanidinoethanesulfonic acid increased the spontaneous release of [3H]taurine by approximately 40%. These results suggest that the taurine release of [3H]taurine is not simply a reversal of the carrier-mediated uptake system. It also appears that taurine is not released from vesicles within the synaptosomes but does not rule out the possibility that taurine is a neurotransmitter. The data involving chloride substitution with permeant and impermeant anions support the concept that the major portion of [3H]taurine release is due to an osmoregulatory action of taurine while depolarization accounts for only a small portion of [3H]taurine release.

4-Aminopyridine↗

PubMed1993

Basal synthesis of acetylcholine in hippocampal synaptosomes is not dependent upon membrane-bound choline acetyltransferase activity.

Choline acetyltransferase, the enzyme which catalyses the formation of acetylcholine within cholinergic nerve terminals, exists in both cytosolic and membrane-associated subcellular pools. In the present study, alteration in nerve terminal Cl- homeostasis was used as an experimental tool to elucidate the role of membrane-bound choline acetyltransferase in regulation of the biosynthesis of acetylcholine in rat hippocampal synaptosomes under basal or resting conditions. Reduction of extracellular Cl- concentration from 131 to 48 mM through iso-osmotic replacement with isethionate ions produced a selective decrease, to approximately 50% of control, of nerve terminal membrane-associated choline acetyltransferase activity. Under these experimental conditions, there were no changes in the activity of cytosolic enzyme or high-affinity choline uptake, or in acetylcholine synthesis. Replacement of medium Cl- with Br- supported maintenance of synaptosomal membrane-bound choline acetyltransferase activity better than did I- or isethionate ions; high-affinity choline uptake activity and acetylcholine synthesis were affected similarly. Incubation of synaptosomes with low concentrations of the Cl- channel blockers 4-acetamido-4'-isothiocyanatostilbene-2,2'-disulphonic acid (50 microM) and niflumic acid (100 microM) selectively decreased activity of the membrane-bound enzyme, with no effect on cytosolic choline acetyltransferase or high-affinity choline uptake activities. Acetylcholine synthesis was unchanged, even though membrane-bound choline acetyltransferase activity was decreased in some samples (250 microM 4-acetamido-4'-isothiocyanatostilbene-2,2'-disulphonic acid) to about 10% of control. Experimental manipulations designed to alter neuronal Cl- homeostasis resulted in selective changes in membrane-bound choline acetyltransferase activity, thereby allowing the first direct examination of its physiological role in regulation of acetylcholine synthesis.(ABSTRACT TRUNCATED AT 250 WORDS)

4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfo↗

PubMed1994

Ca(2+)-dependent Cl- conductance in taste cells from Necturus.

1. Taste responses adapt to a constant chemical stimulus. The present study describes a new ionic conductance in taste cells--a Ca(2+)-dependent anion conductance that may explain taste adaptation. 2. Patch-clamp recordings were made on isolated Necturus taste cells or on taste cells in lingual slices. When Na+ and K+ currents were eliminated with tetrodotoxin (TTX) and tetraethyl-ammonium (TEA) in the bath and replacing K+ with N-methyl-D-glucamine (NMDG+) in the pipette, Ca2+ currents were followed by prolonged outward currents. Outward current was abolished when Ca2+ was substituted with Ba2+ or when Cl- was replaced with large organic anions (methanesulfonate, isethionate, or ascorbate). 3. The outward, Ca-dependent current was reduced by certain agents that block Cl- conductances in other tissues, namely 4-acet-amido-4-isothiocyanostilbene-2,2-disulfonic acid (SITS) and 4,4-diisothiocyanostilbene-2,2-disulfonic acid (DIDS). However, other Cl- channel blockers--9-AC, furosemide and an antibody to Cl channels-had little or no specific effect on the Ca-dependent outward current in Necturus taste cells. 4. We postulate that the biological action of this Ca-dependent anion conductance in situ is to terminate depolarizing receptor potentials, even during maintained chemostimulation, thereby playing an important role in chemosensory adaptation and modulation of impulse discharge patterns in taste buds.

Afferent Pathways↗

PubMed1983

The role of chemiosmotic lysis in the exocytotic release of insulin.

The role of chemiosmotic lysis in the exocytotic release of insulin has been studied using perifused rat pancreatic islets of Langerhans. Established criteria for osmotic lysis of secretory granules requires proton translocation across the secretory granule membrane and the influx of a permeant anion. The consequent increase in granule osmolarity induces water entry and granule lysis. A proton gradient has been previously established to exist across the insulin secretory granule membrane. We have examined the sensitivity of insulin release to 1) hyperosmolar solutions, 2) replacement of medium Cl-, 3) replacement of medium Na+, and 4) anion transport inhibitors. The addition of 200-600 mM sucrose resulted in a 32-69% inhibition of insulin release due to 16.7 mM glucose. Replacement of Cl- by isethionate or SO4--reversibly inhibited glucose-induced insulin release by 47% and 78%, respectively. Na+ replacement by choline did not influence the secretory response. 4,4'-Diisothiocyano-2,2'-stilbene disulfonic acid (500 microM) and probenecid (10 mM) inhibited insulin release by 73% and 79%, respectively. These drugs are known to inhibit anion exchange in erythrocytes and may be influencing Cl- entry into the secretory granule fused to the plasma membrane by a similar mechanism. Furosemide inhibits NaKCl2 cotransport in erythrocytes, but had no influence on glucose-induced insulin release, suggesting that Cl- does not enter the secretory granule by this pathway. The primary criteria for the participation of a chemiosmotic mechanism subserving lysis of the insulin secretory granule are fulfilled by these results.

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

PubMed1988

A large conductance Cl- channel revealed by patch-recordings in human fibroblasts.

A Cl- channel with large single-unit conductance and characteristic voltage-dependent inactivation was studied on cultured human fibroblasts. The channel was activated only after excision and lasting depolarization of the membrane patch. In inside-out configuration and in symmetrical 135 mM NaCl, the conductance was 300 pS. The channel was usually open at the membrane potentials between -20 to +20 mV, while more negative or positive voltages closed the channel. The time course of this apparent inactivation process was dependent on increasing potential. Recovery from inactivation was made possible by returning the membrane potential to 0 mV. The channel was selective to Cl- over Na+ with a PCl/PNa of 6. The order of permeability among anions was: I greater than Br = Cl greater than isethionate greater than F greater than glutamate. The channel was blocked by internal application of a derivative of the diphenylamine-2-carboxilate (Blocker 144) but not by 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid.

Chlorides↗

PubMed1999

Participation of a chloride conductance in the subthreshold behavior of the rat sympathetic neuron.

The presence of a novel voltage-dependent chloride current, active in the subthreshold range of membrane potential, was detected in the mature and intact rat sympathetic neuron in vitro by using the two-microelectrode voltage-clamp technique. Hyperpolarizing voltage steps applied to a neuron held at -40/-50 mV elicited inward currents, whose initial magnitude displayed a linear instantaneous current-voltage (I-V) relationship; afterward, the currents decayed exponentially with a single voltage-dependent time constant (63.5 s at -40 mV; 10.8 s at -130 mV). The cell input conductance decreased during the command step with the same time course as the current. On returning to the holding potential, the ensuing outward currents were accompanied by a slow increase in input conductance toward the initial values; the inward charge movement during the transient ON response (a mean of 76 nC in 8 neurons stepped from -50 to -90 mV) was completely balanced by outward charge displacement during the OFF response. The chloride movements accompanying voltage modifications were studied by estimating the chloride equilibrium potential (E(Cl)) at different holding potentials from the reversal of GABA evoked currents. [Cl(-)](i) was strongly affected by membrane potential, and at steady state it was systematically higher than expected from passive ion distribution. The transient current was blocked by substitution of isethionate for chloride and by Cl(-) channel blockers (9AC and DIDS). It proved insensitive to K(+) channel blockers, external Cd(2+), intracellular Ca(2+) chelators [bis-(o-aminophenoxy)-N,N,N',N'-tetraacetic acid (BAPTA)] and reduction of [Na(+)](e). It is concluded that membrane potential shifts elicit a chloride current that reflects readjustment of [Cl(-)](i). The cell input conductance was measured over the -40/-120-mV voltage range, in control medium, and under conditions in which either the chloride or the potassium current was blocked. A mix of chloride, potassium, and leakage conductances was detected at all potentials. The leakage component was voltage independent and constant at approximately 14 nS. Conversely, gCl decreased with hyperpolarization (80 nS at -40 mV, undetectable below -110 mV), whereas gK displayed a maximum at -80 mV (55.3 nS). Thus the ratio gCl/gK continuously varied with membrane polarization (2.72 at -50 mV; 0.33 at -110 mV). These data were forced in a model of the three current components here described, which accurately simulates the behavior observed in the "resting" neuron during membrane migrations in the subthreshold potential range, thereby confirming that active K and Cl conductances contribute to the genesis of membrane potential and possibly to the control of neuronal excitability.

Animals↗

PubMed1990

The anion in salt taste: a possible role for paracellular pathways.

It is well established from psychophysical and electrophysiological measurements that both Na and Cl contribute to the taste response to NaCl. The contribution of Na to the NaCl response can be studied using amiloride, a drug that inhibits Na transport in taste and other epithelial cells. The pathways involved in response to Cl are less well understood. We undertook a series of experiments in the rat to determine whether tonic chorda tympani responses to NaCl are inhibited by specific inhibitors of anion transport. Whole nerve responses to NaCl were unchanged by bathing the tongue in SITS, DIDS, bumetanide, furosemide, 9-anthracene carboxylic acid, or an antibody that blocks Cl conductance pathways in many epithelia. Thus, Cl co-transporters, exchangers, and channels (at least in the apical membrane of taste cells) are probably not involved in NaCl taste responses. When other anions (acetate, isethionate, methane sulfonate, gluconate, tartrate), which are generally impermeant in other Cl-selective pathways, were substituted for Cl, the dose-response curves for the chorda tympani response were shifted toward higher concentrations than the response to NaCl, but achieved the same maximum value at sufficiently high concentrations (1.0 M Na). For all the organic Na salts, the amiloride-insensitive portion of the response was substantially less than for NaCl. Experiments with Na acetate at different pHs showed that intracellular acidification is not responsible for the differences between NaCl and organic salts of Na. One possibility which remains is that apical stimulation with these other Na salts results in a taste cell membrane potential that is hyperpolarized with respect to the membrane potential in NaCl.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetates↗

PubMed1994

Calcium-dependent chloride current induced by axotomy in rat sympathetic neurons.

1. Seven to ten days after sectioning their axons, rat sympathetic neurons were studied using intracellular recording techniques in an in vitro preparation of the superior cervical ganglion. 2. In 75% of axotomized cells, an after-depolarization (ADP) was observed following spike firing or depolarization with intracellular current pulses. Discontinuous single-electrode voltage-clamp techniques were employed to study the ADP. When the membrane potential was clamped at the resting level just after an action potential, a slow inward current was recorded in cells that showed an ADP. 3. In the presence of TTX and TEA, inward peaks and outward currents were recorded during depolarizing voltage jumps, followed by slowly decaying inward tail currents accompanied by large increases in membrane conductance. The inward peak and tail currents activated between -10 and -20 mV and reached maximum amplitudes around 0 mV. With depolarizing jumps to between +40 and +50 mV, net outward currents were recorded during the depolarizing jumps but inward tail currents were still activated. 4. In the presence of the Ca2+ channel blocker cadmium, or when Ca2+ was substituted by Mg2+, the ADP disappeared. In voltage-clamped cells, cadmium blocked the inward tail currents. The reversal potential for the inward tail current was approximately -15 mV. Substitution of the extracellular NaCl by sucrose or sodium isethionate increased the amplitude of the inward tail current, and displaced its equilibrium potential to more positive values. Changes in extracellular [K+] did not appreciably affect the inward tail current amplitude or equilibrium potential. Niflumic acid, a blocker of chloride channels activated by Ca2+, almost completely blocked the tail current. 5. No ADPs were observed in non-axotomized neurons, and when depolarizing pulses were applied while in voltage clamp no inward tail currents were evoked in these normal cells. 6. It is concluded that axotomy of sympathetic ganglion cells produces the appearance of a Ca(2+)-dependent chloride current responsible for the ADP observed following spike firing.

Animals↗

PubMed2004

Bradykinin decreases K(+) and increases Cl(-) conductances in vagal afferent neurones of the guinea pig.

Bradykinin (BK) is an inflammatory mediator that can excite and sensitize primary afferent neurones. The nature of the ionic channels underlying the excitatory actions of BK is still incompletely understood. Using whole-cell patch-clamp recording from acutely dissociated nodose ganglion neurones (NGNs) we have examined the ionic mechanism responsible for BK's excitatory effect. Bath-applied BK (0.1 microM) depolarized the membrane potential (29 +/- 3.1 mV, n= 7), evoked action potentials, and induced an inward ionic current (I(BK)) with two distinctive membrane conductances (g(m)). Initially, g(m) decreased; the ionic current associated with this g(m) had a reversal potential (E(rev)) value of -87 +/- 1.1 mV (n= 26), a value close to E(K) (-89 mV). Subsequently, g(m) increased; the ionic current associated with this g(m) had an estimated E(rev) of 49 +/- 4.3 mV (n= 23). When the second component was isolated from the first component, by replacing [K(+)](o) with Cs(+), E(rev) was 20 +/- 4.7 mV (n= 10). Replacing external NaCl with NMDG-Cl or choline-Cl, or reducing [Ca(2+)](o) did not significantly diminish I(BK). After replacing external NaCl with sodium isethionate, E(rev) for the second component shifted to 56 +/- 8.8 mV (n= 4), a value close to the E(Cl) (66 mV). The second component was inhibited by intracellular BAPTA or by bath application of niflumic acid (100 microM), a Ca(2+)-activated Cl(-) channel blocker. These results suggest that the first and second components of I(BK) are produced by a decrease in K(+) conductance and an increase in Ca(2+)-activated Cl(-) conductance, respectively. The BK-evoked Cl(-) conductance in NGNs may be the first demonstration of an inflammatory mediator exciting primary afferents via an anion channel.

Animals↗