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Effects of hypertonic solution on action potential and input resistance in the guinea-pig ventricular muscle.

The effects of hypertonic solution on action potential and input resistance were studied using the guinea-pig ventricular muscle. Hypertonic solutions containing excess (150 and 300 mOsm/liter) sucrose, glucose, NaCl, Na2SO4, or LiCl produced a prolongation of action potential duration (APD) followed by a gradual shortening. These solutions increased the input resistance of the preparation. The prolongation of APD began immediately after the onset of perfusion with hypertonic solution and peaked within about 10 min, while the late shortening of APD developed progressively. On introducing the isotonic solution after the osmotic challenge, APD initially shortened and then gradually prolonged toward the control level. Slightly hypertonic solutions containing excess 30 or 75 mM sucrose prolonged APD, to some extent. It is concluded that the hypertonic solution exerts early and late effects on APD in guinea-pig ventricular muscle. The early effect, which is responsible for the APD prolongation, seems to be directly related to the development of cell dehydration. The hypertonic solutions produced a slight increase in the resting potential, and sucrose- and glucose-hypertonicity depressed the rising phase of the action potential. These effects can be explained by the cell dehydration and associated changes in the intracellular ion concentrations. The increase in input resistance produced by hypertonic solutions was accompanied by an enhanced spatial decay of electrotonic potentials along the muscle bundle, as determined by the two microelectrode method, suggesting that the electrical resistance of the internal current pathway increases under hypertonic conditions.

Action Potentials↗

The membrane properties of the smooth muscle of the guinea-pig portal vein in isotonic and hypertonic solutions.

The membrane properties of the longitudinal smooth muscle of the guinea-pig portal vein were investigated under various experimental conditions.1. In isotonic Krebs solution, the membrane potential (-48.7 mV), the maximum rates of rise and fall of the spike (4.6 and 2.3 V/sec respectively), the space constant (0.61 mm), the conduction velocity of excitation (0.97 cm/sec) and the time constant of the foot of the propagated spike (18.4 msec) were measured.2. The various parameters of the muscle membrane in the isotonic solution were compared with those in the hypertonic solution prepared by the addition of solid sucrose (twice the normal tonicity).3. When the muscles were perfused with hypertonic solution, marked depolarization of the membrane and increased membrane resistance occurred. These were probably due to reduction of the K permeability, increased internal resistance of the muscle and shrinkage of the muscle fibre.4. The membrane potential in isotonic and hypertonic solutions was analysed into two components, i.e. the metabolic (electrogenic Na-pump) and the ionic (electrical diffusion potential) component in the various environmental conditions.(a) In isotonic and hypertonic solutions, the membrane was depolarized by lowering the temperature or by removal of K ion from the solutions. When the tissues were rewarmed or on readdition of K ion, the membrane was markedly hyperpolarized. These hyperpolarizations of the membrane were suppressed by treatment with ouabain (10(-5) g/ml.), by warming to only 20 degrees C and by K-free solution.(b) The relationships between the membrane potential and the [K](o) in isotonic Krebs, in the hypertonic (sucrose) Krebs, in the Na-free (Tris) Krebs and in the Cl-deficient (C(6)H(5)SO(3)) Krebs were observed. The maximum slopes of the membrane depolarization against tenfold changes of [K](o) were much lower than that expected if it behaved like a K electrode.(c) In Na-free (Tris) solution, the membrane was not depolarized in isotonic condition but it was depolarized in hypertonic condition.5. The low membrane potential in hypertonic solution (-37 mV) compared with isotonic solution (-49 mV) was thought to be mainly due to suppression of K permeability of the membrane and not due to suppression of the metabolic component. The electrogenic Na-pump and the membrane potential of the portal vein was discussed in relation to other excitable cell membranes.

Action Potentials↗

Therapeutic efficacy of 5% NaCl hypertonic solution in patients with bullous keratopathy.

A clinical trial was undertaken to evaluate the efficacy of hypertonic solution (5% NaCl) in patients who have bullous keratopathy (BK). The aim of the study was to define the stage of the disease and the thickness of cornea in micrometers, which would be the threshold for therapeutic approach. This was a prospective study on 70 eyes of 55 patients. Patients were divided in two groups at the beginning of the study. The first group (n=33 eyes) included patients with initial stage of BK: only stromal component of corneal oedema was present. The second group (n=37 eyes) included patients with advanced stage of BK: the epithelial component of the disease with bullae on the corneal surface had already developed. Visual acuity, central and peripheral thickness of cornea and morphology of the disease was recorded before therapy, 7 days and 4 weeks after administration of hypertonic solution. Our results shown that the efficacy of hypertonic solution correlates with the severity of clinical picture in patients with BK. When 5% NaCl hypertonic solution was applied in the early stage of the disease, when only stromal component of corneal oedema was presented, visual acuity and pachymetry readings were significantly improved. The threshold pachymerty measurement of corneal thickness justifying the application of hypertonic solution was 613-694 microm (in the central corneal area), and 633-728 microm (at corneal periphery). It seems reasonable to apply hypertonic solution to the patients who have BK and whose pachymetric values are below mentioned range. In terminal stages of BK, when superficial bullae (epithelial component) had already developed, treatment with NaCl was not effective and patients had to be submitted to penetrating keratoplasty.

Aged↗

Effects of hypertonic solutions on calcium transients in frog twitch muscle fibres.

1. The effects of hypertonic solutions on excitation-contraction (e.-c.) coupling in frog skeletal muscle fibres were investigated using Arsenazo III to monitor intracellular calcium transients in voltage-clamped fibres. 2. In solutions made hypertonic with sucrose or sodium chloride, the size of the Arsenazo signal evoked by a 5 ms depolarization to 0 mV was little altered by increases in tonicity up to about twice normal, but declined in higher tonicities, and was almost completely suppressed at 4 times normal tonicity. 3. The latency to onset of the Arsenazo signal was increased in hypertonic solutions (2.3 and 3.1 times normal tonicity), but the decay time constant of the signal was little changed with tonicities up to 2.3 times normal. 4. The rheobase potential for a just-detectable Arsenazo signal was shifted about 4 mV more negative by increases in tonicity up to 2.3 times normal, but further increases reversed the direction of the shift, and in 3.95 times normal tonicity the rheobase was 10 mV more positive than in normal Ringer solution. 5. With short (less than 10 ms) pulse durations the depolarization needed to elicit a threshold Arsenazo signal increased steeply with increasing tonicity. Changes in the strength-duration curve could be accounted for by an increase in the time constant for build-up of a hypothetical coupler in the e.-c. coupling process. 6. Solutions of about twice normal tonicity are commonly used to suppress muscle contraction. Since the size of the Arsenazo signal was only slightly reduced by this tonicity, the main effect is presumably on the contractile proteins.

Animals↗

T-tubule swelling in hypertonic solutions: a freeze substitution study.

Striated muscles from Rana pipiens have been exposed for variable periods of time to Ringer solutions made hypertonic by addition of either sucrose or sodium chloride. The muscles have been rapid-frozen and then prepared for electron microscopy by either freeze-substitution, freeze-fracture or cryoultramicrotomy. The only compartment greatly affected by hypertonicity is the transverse tubular system, which is visibly swollen. None of the elements of the sarcoplasmic reticulum increase in size.

Animals↗

Mucus glycoconjugate secretion in cool and hypertonic solutions.

For sensitive individuals, exercise-induced asthma is triggered by cold and dry air and is often accompanied by a productive cough. In this study, we determined whether cold solutions and/or solutions of increased tonicity directly caused an increase in glycoconjugate (GC) secretion. To test this, we used isolated swine tracheal submucosal gland cells (TSGCs) and measured the rate of GC secretion at 37 and 32 degrees C in isotonic solutions and in solutions made hypertonic by 30 mosM. TSGCs were isolated under conditions that minimized the rate of GC secretion and were perfused with medium 199 equilibrated with 5% CO2 to a pH of 7.4. A lectin-based assay was used to specifically detect GC present in each 2-min fraction of the perfusate. Basal secretion was 3.1-fold greater at 32 degrees C (n = 3) than at 37 degrees C (n = 4; P < 0.05). At 37 degrees C, increasing perfusate osmolarity by 30 mosM increased the average rate of secretion by 41 +/- 11% (n = 4; P < 0.05); return to isotonic perfusate caused a 4.5 +/- 1.8-fold transient increase in secretion (n = 4; P < 0.05) that was complete within 10 min. At 32 degrees C, changing tonicity of the perfusate had no significant effect but returning to isotonic perfusate caused a 2.3 +/- 0.7-fold transient increase in secretion (n = 3; P < 0.05). Thus key stimuli that trigger obstruction of airflow (cold and increased osmolarity) can also directly stimulate GC secretion in the airway. Such increased secretions may contribute to the productive cough observed in some individuals in response to cold air.

Animals↗

Analysis of the effluxes of sodium, potassium and chloride ions from smooth muscle in normal and hypertonic solutions.

1. Efflux curves of (24)Na, (42)K and (36)Cl from the guinea-pig taenia coli were obtained in normal Krebs solution, and in hypertonic Krebs solution in which the osmolarity had been doubled by the addition of sucrose.2. The efflux curves were complex, and in order to get average curves each was analysed as the sum of three exponential terms, and average curves were constructed from the means of the constants found.3. In order to estimate the membrane permeability to the ions, it was necessary to make assumptions as to the distribution of ions in the tissue. Several models have been examined and the predictions of the models with respect to the membrane properties were compared with the data obtained by using electrophysiological methods by other workers.4. It was found that reasonable predictions of membrane properties could only be made using models in which the majority of the rapidly exchanging sodium is considered to be extracellular. This amount of sodium is more than can be accounted for as freely dissolved in the extracellular water.5. A possible interpretation of the ion exchange and distribution would be to suppose that some proportion of the three ions is contained in an extracellular compartment not available to the normal extracellular markers, and limited in its exchange by the rate of diffusion in the extracellular phase, and that the truly intracellular fractions of the tissue ions do not exchange with the external solution in a simple exponential manner, but in a manner described by an aggregate of exponential terms due to inherent variation in the permeabilities of the individual cell membranes.6. There is no evidence for any change in the membrane permeabilities to sodium and potassium in hypertonic solution, but there is evidence for a decrease in chloride permeability in this solution.

Animals↗

Inhibition by hypertonic solutions of Ca-dependent electrogenesis in single crab muscle fibers.

This study describes the effect of hypertonic solutions on isolated muscle fibers of Callinectes danae. Solutions of twice normal tonicity (2.0 T) inhibit both the normal graded membrane responses and the spikes induced by procaine, tetraethylammonium, or barium. The inhibition is maintained throughout exposure to hypertonic solutions prepared by addition of impermeant solutes such as NaCl, sucrose, or Tris-propionate, but is reversible on their withdrawal. In the presence of permeant solutes such as glycerol or acetamide, the inhibition is transient. In both cases the onset of inhibition of the depolarizing Ca electrogenesis is correlated with shrinkage of the fiber. In the case of permeant solutes, the time course of recovery of the graded responses or the spikes follows the recovery of the fiber volume. Changes in the passive electrical characteristics of the fibers due to hypertonic solutions were unrelated to the blockade of membrane Ca activation. The current-voltage relationship in hypertonic sollution revealed no increase in depolarizing K activation. Inhibition of the graded membrane responses and spikes appears to be associated with depression of Ca conductance. Hypertonic solutions might affect the activation of Ca conductance through reduction of the electric field generated by fixed negative surface charges and/or morphological changes in the T tubules. Membrane depolarization elicited little or no tension in 2.0 T solutions while caffeine contracture (10 mM) with an ampliture of 76% of the maximal contractile ability could still be elicited. This indicates that direct effects of hypertonic solutions on the contractile apparatus were not responsible for loss of tension. The latter is attributed to the inhibition of the transmembrane Ca currents.

Acetamides↗

The effect of hypertonic solution on the wet weight and contractions of rat uterus and vas deferens.

The role of propagated activity in the responses to agonist drugs was studied for the rat uterus and vas deferens. Hypertonic solutions were used to inhibit propagation of activity by shrinking cells. Tissue weight was used to indicate cell volume. Hypertonic solutions after 10 min caused weight loss and reduced the size of contractions in response to submaximal doses of drugs, to KCl, and to external electrical stimulation. Contractions in response to KCl and drugs were diminished to a similar degree in the vas deferens, but in the uterus, drug contractions were depressed much more. Prolonged action of hypertonic solution also differed for the two tissues. In the uterus, weight changes correlated with changes in size of the drug-induced contractions. Uterine contractions reduced in hypertonic solution could be increased by using supramaximal doses of drug. When stimulation was applied to one end of the uterus in a three compartment bath, propagation of spontaneous drug- and KCl-induced contraction occurred, but it was prevented by placing hypertonic solution in the center compartment. An increase of the KCl to 44 mM in the hypertonic solution restored propagation. These experiments yielded no evidence of propagated responses in the rat vas deferens. It was concluded that propagated activity plays a role in drug-induced contractions in the rat uterus but not in the rat vas deferens. Hyperpolarization of shrunken cells might be involved in inhibition of propagation by hypertonic solutions.

Acetylcholine↗

Enhancement of aorta tonus evoked by hypertonic solutions is endothelium-dependent.

1. The effect of NaCl hypertonic solutions on the contractions induced by noradrenaline or prostaglandin F2 alpha and on the relaxant effect of acetylcholine and sodium nitroprusside was investigated in aortae with or without endothelium isolated from rats. 2. Hypertonic solutions enhanced the contractions elicited by the vasoconstrictor agents in preparations with endothelium and this phenomenon was not altered by previous treatment of the animals with reserpine or the preparations with methylene blue or indomethacin. 3. In hypertonic medium acetylcholine but not sodium nitroprusside vasodilator effect was inhibited. 4. We suggest that NaCl hypertonic solutions modify vascular reactivity probably through: (i) the release of an endothelium-derived contracting factor not sensitive to cyclooxygenase inhibitors; (ii) a diminished release of an endothelium-derived relaxing factor not related to the cGMP system.

Acetylcholine↗

Antiulcer activity of hypertonic solutions in the rat: possible role of prostaglandins.

The effects of hypertonic solutions on gastric acidity and on experimental gastric and duodenal ulcers as well as on gastric prostaglandins (PGs) were studied in the rat. The oral administration of a 10% NaCl solution resulted in complete absence of free acidity and very significant reductions in total acidity 24 h after pyloric ligation. The antiulcer effect of hypertonic saline was studied in three experimental models. In pyloric-ligated rats, both the incidence and the severity of gastric ulcers were remarkably reduced by hypertonic saline treatment. Indomethacin-induced gastric erosions were significantly reduced by hypertonic NaCl or sorbitol and completely prevented by hypertonic xylitol. Cysteamine-induced duodenal ulcers were also significantly reduced by hypertonic solutions of NaCl, xylitol or sorbitol. In the latter model, indomethacin potentiated the ulcerogenic effect of cysteamine and also reduced the efficacy of the hypertonic NaCl gavage. The possible contribution of PGs to these effects was further investigated by analysing PGE in the gastric mucosa and juice. Rats treated orally with hypertonic NaCl solutions had several-fold higher PGE contents in their gastric mucosa as well as higher PGE levels in the gastric juice. It is concluded that hypertonic solutions stimulate endogenous PGE biosynthesis and also exert profound antiulcer effects in the rat. A causal relationship between the two phenomena is suggested.

Animals↗

The increase in the rate of heat production of frog's skeletal muscle caused by hypertonic solutions.

1. The rate of heat production of resting muscle is increased by hypertonic solutions.2. The threshold osmolality required to produce the increased heat rate is less than 2 times normal; at 2.5-3 times normal the heat production rises to 20-50 mcal.g(-1).min(-1), which is 10-20 times the basal rate.3. In anaerobic conditions, the effect of hypertonic solutions on heat rate is only one tenth of that in aerobic conditions.4. A glycerol-treated muscle, with damaged tubular system, still gives a normal response to hypertonic solutions, though it does not respond to raised K(+) concentration.5. The metabolic response to hypertonic solutions is considerably suppressed by procaine.6. Ouabain, 10(-5)-10(-4)M, has no effect.7. The response remains substantial in a muscle which has been depolarized in isotonic K(2)SO(4).8. The membrane potential is slightly reduced by hypertonic solutions, but this cannot account for the increase of the resting metabolism.9. It is suggested that the effect may be due to the release of calcium ions, which produce an increase in myosin ATPase activity.

Adenosine Triphosphatases↗

Effects of hypertonic solutions on conjunctival epithelium and mucinlike glycoprotein discharge.

A simple, semiquantitative, histochemical method using periodic acid-Schiff (PAS) staining was developed to measure the mucinlike glycoprotein content of bathing solutions exposed to rabbit conjunctiva. Rabbit eyes were bathed in vivo in solutions of various tonicity (160, 260, 300, 330, and 363 mOsm/L). Eyes bathed in hypotonic or isotonic solutions (160, 260, and 300 mOsm/L) exhibited a steady-state glycoprotein secretion (50-100 micrograms/h). In hypertonic solutions (330 and 363 mOsm/L), glycoprotein discharge increased to 150-360 micrograms/h when the solutions were exchanged hourly for 6 h, but glycoprotein discharge increased to a much lower rate (100-167 micrograms/h) after prolonged bathing for 6 h without solution exchange. Periodic acid-Schiff--positive glycoprotein aggregates were noted more frequently on the filters of hypertonic solutions than on those of other tonicity. After eyes were bathed in hypertonic solutions, increased disruption of the surface epithelium, increased abnormal discharge of mucin granules, and decreased goblet cell density were noted in the biopsied conjunctiva. These data suggest that mucinlike glycoprotein secretion from the conjunctival goblet cells can be enhanced by the tonicity of the bathing solution, and that nonphysiological hypertonic solutions can also affect the conjunctival epithelium.

Animals↗

[Hypertonic solutions in treatment of intracranial pressure].

Administration of hypertonic solutions is the method of choice for acute treatment of intracranial hypertension. Recording of the intracranial pressure during treatment facilitates adjustment of the dosis to the actual ICP-response, avoiding thereby administration of an excessive osmotic load as a basis to prolong therapeutical efficacy. The mechanisms underlying reduction of the intracranial pressure by hypertonic solutions are still controversially discussed. Dehydration of normal probably also of edematous brain parenchyma and constriction of cerebral resistance vessels as an autoregulatory response causing reduction of the intracranial blood volume are the most likely options. Administration of hypertonic/hyperoncotic solutions has regained attention on account of its unmatched therapeutical efficacy to reestablish normal conditions in severe hemorrhagic shock. Administration of, e.g. 7.2% NaCl/10% Dextran 60 in an amount equivalent of only 10% of the shed blood volume is immediately normalizing cardiac output and improving the microcirculation in peripheral organs. These therapeutical properties are relevant in head injury, since inflicted patients quite often are suffering from peripheral trauma and consequently from hemorrhagic shock. No evidence has been obtained in a variety of experimental studies that hypertonic/hyperoncotic solutions have adverse effects on the brain in the presence of a cerebral lesion. To the contrary, the fluid mixture has been found to lower the increased intracranial pressure. Administration of hypertonic/hyperoncotic solutions appears therefore appropriate in acute cerebral insults from head injury and impending circulatory failure from shock in order to inhibit development of secondary brain damage.

Animals↗

Effects of hypertonic solutions on quantal transmitter release at the crayfish neuromuscular junction.

1. The effects of a hypertonic bathing medium, containing either NaCl or melezitose, on the average number (m) and the time course of quantal release following an action potential were studied using focal extracellular recording methods at synaptic sites on the opener muscle of the crayfish leg.2. After the application of hypertonic saline, the rate of spontaneous quantal release increased but m decreased to a new level within 1 min; the extent of depression depended on the magnitude of the increase in tonicity until the osmolarity was 50% greater than the normal value of 0.43 osmol/l (Osm), but greater increases in tonicity exerted little further effect.3. The synaptic delays were increased and distributed over a longer range of time in hypertonic solutions; also, the latency between the first and second quantal releases in a multiple response to a single nerve impulse was also increased.4. Hypertonicity had no significant effect on the conduction velocity of the action potential, the independence of successive quantal releases in the same response, or the uniformity of the rise and fall of the probability of quanta release following an action potential, alpha(t).5. The time course of alpha(t) is prolonged in hypertonic solutions; this was manifested as an increase in the time constant of the exponential decline in alpha(t) from its peak value following the nerve impulse.5. When the potentiating agent 5-hydroxytryptamine (5-HT) was added to the hypertonic saline, m was increased, and the time course of alpha(t) was prolonged further than in the hypertonic solution alone; 5-HT produced no change in the time course of alpha(t) when it was added to normal saline, although m was increased.6. It is concluded that in this preparation hypertonicity decreases the rate of release of each quantum from the nerve terminal following a nerve impulse.

Action Potentials↗

Water drinking caused by intracerebroventricular infusions of hypertonic solutions in cattle.

Infusions into the lateral cerebral ventricle of hypertonic solutions of NaCl, mannitol or sucrose all induced water drinking in cattle. However, infusion of hypertonic NaCl caused a significantly greater water drinking response than did the infusions of mannitol or sucrose, despite the fact that CSF osmolality increase was similar. In contrast, hypertonic solutions of NaCl or mannitol had similar dipsogenic effects when infused intravenously. The intracerebroventricular infusions of hypertonic NaCl or mannitol did not affect the intakes of food or Na solution. The results are consistent with the hypothesis that both cerebral osmoreceptors and Na sensors are involved in regulating thirst in cows.

Animals↗