Influence of short-chain fatty acids on ammonia absorption across the rumen wall in sheep.

The effects of short-chain fatty acids (SCFA) on ammonia net absorption from the sheep rumen in vivo and on ammonia transport across rumen wall mucosa in vitro were studied. Ammonia net absorption was directly, though in a non-linear manner, correlated with the SCFA concentration in the artificial rumen fluid. Almost 70% of total ammonia absorption was dependent upon the presence of SCFA when 12 mmol l-1 ammonia and 67.5 mmol l-1 SCFA were present. Lactic acid was ineffective. Incubation experiments showed that mucosal disappearance and serosal appearance of ammonia were reduced by 38% and 32%, respectively, when SCFA (63 mmol l-1) were replaced by lactic acid. The SCFA effect was independent of the type of SCFA used. In part of the experiments up to 54% of the ammonia taken up by the tissue was not recovered in the serosal incubation solution and must have been metabolized in the mucosa.     

The effect of a rumen microbial fermentation on urea and nitrogen metabolism of sheep nourished by intragastric infusion

Four sheep were maintained by infusion of volatile fatty acid (VFA), mineral and buffer solutions to the rumen and casein to the abomasum. After a 3 week period in which control measurements were made, glucose was introduced gradually to the rumen infusion mixture until glucose replaced about 65% of VFA energy. Further measurements were made in the fourth week of glucose additions after which the control VFA infusions were re-established and measurements repeated. Urea kinetics were measured with [14C]urea. Rumen ATP concentrations were used as an index of microbial growth and increased from 1.5 nmol ml-1 on control treatments to 46 nmol ml-1 when glucose was given. The presence of a rumen fermentation resulted in a 40% decrease in plasma urea concentration but was without effect on rumen ammonia concentration. Urea irreversible loss rate decreased from 18.0 to 12.8 g urea day-1 when glucose was given. This, however, was matched by a decrease in urinary urea excretion, with the result that urea degradation, obtained by difference, remained constant throughout the three treatment periods. Daily nitrogen (N) retention increased from 1.0 to 2.7 g when a rumen microbial population was present. A model of urea and NH3 transactions for the two dietary situations is presented. Calculations indicated that 0.40 of the casein N supply was reutilized as urea by the rumen micro-organisms. It is suggested, however, that the improvement in N retention resulted from a change in protein:energy ratio of the infused nutrients rather than from an enhanced supply of protein to the intestine. The use of endogenous urea clearance as an index of epithelial permeability to urea is questioned.     

Transport of calcium in the perfused submandibular gland of the cat

1. In the perfused cat submandibular gland efflux and influx of (45)Ca, and concentrations of K, (40)Ca and Mg in the effluent from the gland were measured under different experimental conditions.2. When the standard perfusion fluid was shifted to a high Mg (5 mM) or a low Ca (0.25 mM) solution the efflux of (45)Ca from the pre-labelled gland declined. The magnitude and the duration of the effect of the high Mg concentration was more marked at a low external Ca concentration and was abolished by Mersalyl (1 mM). When the standard perfusion fluid was shifted to a Mg-free solution the efflux of (45)Ca from the pre-labelled gland increased.3. After shift of (45)Ca containing perfusion fluid from normal to a high Mg (5 mM) solution the influx of (45)Ca to the gland increased rapidly.4. Both acetylcholine (ACh) and adrenaline caused a marked increase in the efflux of (45)Ca from the pre-labelled gland. This increase in efflux was also seen under conditions where the gland was unable to secrete, i.e. during perfusion with Ca-free and Na-free tetraethylammonium Locke solutions.5. Stimulation with ACh failed to reveal any rapidly occurring increase in influx of (45)Ca.6. Stimulation with ACh evoked a small temporary increase in the concentration of (40)Ca. and Mg in the effluent.7. It is suggested that Ca uptake by intracellular Ca-accumulating systems of the submandibular gland depends on the external Mg concentration and that ACh and adrenaline cause a release of Ca bound intracellularly.     

Water and sodium movements across the ruminal epithelium in fed and food-deprived sheep

Fluid and electrolyte movements across the ruminal epithelium of sheep were studied using the temporarily isolated rumen technique. The sheep were all subjected to the following treatments: (1) fed sheep (fed twice daily), after rumen emptying, received rumen contents from a fed sheep; (2) food-deprived sheep (two meals were omitted), after rumen emptying, received rumen contents from a food-deprived sheep; (3) fed sheep, after rumen emptying, received rumen contents from a food-deprived sheep; and (4) food-deprived sheep, after rumen emptying, received rumen contents from a fed sheep. Food deprivation led to an increased Na concentration of the rumen fluid while K and Cl concentrations, as well as osmolality, decreased. Plasma Na and osmolality decreased. During the 40 min after the rumen contents were exchanged no net movements of water occurred. Then the sheep were given an intraruminal load of saline which gave rise to a significant net absorption of fluid from the rumens of those sheep which had received rumen contents from fed sheep. The change in composition of the rumen contents after food deprivation impaired the absorption of Na and water from the rumen. Furthermore food deprivation reduced the Na absorptive function of the ruminal epithelium, but not the water permeability.     

Observations on the potential across the rumen of the sheep.

1. The electric potential difference between rumen contents and jugular venous blood was measured in anaesthetized sheep. In order to investigate the effect on the potential of changing the ionic concentrations within the rumen, the digesta were removed from the rumen and various salt solutions were substituted. The reticulo-rumen sac was isolated before the experiment by ligation of the oesophagus and the reticulo-omasal junction. 2. The observation of Dobson & Phillipson (1958) that the rumen contents are normally of the order of 30 mV negative to the blood was confirmed. 3. For potassium concentrations between 25 and 100 mM the potential at constant [Na+] varied linearly with log [K+]. With sulphate as the anion, the slope for a 10-fold concentration change was 39.7 +/- 3.0 mV when [Na+] was around 50 mM. The slope showed a tendency to increase when [Na+] was lowered, and to decrease when [Na+] was raised. 4. When chloride was substituted for sulphate, both the slope and the absolute size of the potential were slightly reduced. 5. When the sodium concentration was varied at constant [K+], the potential increased as an approximately linear function of [Na+]. At around 10 mM-K the mean slope was 0-32 +/- 0.07 mV/mM; at the highest potassium concentrations it fell to 0-13 +/- 0 05 mV/mM. 6. In most of these experiments isotonicity was maintained with sucrose. The results of a few tests in which Li+ was substituted for Na+ or K+ suggested that the rumen epithelium behaves in a relatively inert fashion towards this ion.     

Net transport of glucose from blood to cerebrospinal fluid in the cat

The net transport of glucose from blood to the cerebrospinal fluid compartment of cats was measured by ventriculocisternal perfusion to determine over a large range of serum glucose concentrations the influence of serum glucose levels and their changes on the net transport rate. Changes in serum glucose levels were followed within minutes by corresponding changes in cerebroventricular effluent fluid glucose concentration. At mean values of serum glucose concentration of 6.2 mM and cerebrospinal fluid formation rate of 24.3 microliter/min, the net glucose influx rate was 1.6 mmol/min. The effluent fluid-to-serum glucose concentration ratio was 0.25 and decreased when serum glucose was greater than 11.1 mM. The rate of glucose transport from blood to effluent fluid during ventricular perfusion was saturable, and approached a maximum of 3.5 mumol/min at serum glucose levels above 22 mM. From the cerebrospinal fluid formation and net glucose influx rates the calculated glucose concentration of nascent cerebrospinal fluid was 6.5 mM and higher than the corresponding serum glucose of 5.6 mM. It is concluded that during perfusion over a wide range of serum glucose concentrations, a saturable mediated glucose transport mechanism can be demonstrated. Changes in serum glucose are rapidly reflected in corresponding effluent fluid glucose levels. From effluent fluid-to-serum glucose concentration ratios and calculations of the glucose in newly formed cerebrospinal fluid, the technique, however, overestimates the glucose influx rates at normal serum glucose levels.     

Bidirectional substrate fluxes through the System N (SNAT5) glutamine transporter may determine net glutamine flux in rat liver

System N (SNAT3 and SNAT5) amino acid transporters are key mediators of glutamine transport across the plasma membrane of mammalian cell types, including hepatocytes and astrocytes. We demonstrate that SNAT5 shows simultaneous bidirectional glutamine fluxes when overexpressed in Xenopus oocytes. Influx and efflux are both apparently Na⁺ dependent but, since they are not directly coupled, the carrier is capable of mediating net amino acid movement across the cell membrane. The apparent K _m values for glutamine influx and efflux are similar (∼1 m m ) and the transporter behaviour is consistent with a kinetic model in which re-orientation of the carrier from outside- to inside-facing conformations (either empty or substrate loaded) is the limiting step in the transport cycle. In perfused rat liver, the observed relationship between influent (portal) glutamine concentration and net hepatic glutamine flux may be described by a simple kinetic model, assuming the balance between influx and efflux through System N determines net flux, where under physiological conditions efflux is generally saturated owing to high intracellular glutamine concentration. SNAT5 shows a more periportal mRNA distribution than SNAT3 in rat liver, indicating that SNAT5 may have particular importance for modulation of net hepatic glutamine flux.     

Drug uptake into everted intestinal sacs. II. Inhibition of secretion by hypertonicity.

Mucosal hypertonicity, metabolic inhibitors, or absence of glucose and oxygen enhance mucosal-to-serosal influx of the cationic drug, pralidoxime (PAM), into sacs of everted rat jejunum in vitro. Conversely, efflux of PAM, which is twice the influx rate, is inhibited by mucosal hypertonicity or cyanide and iodoacetate. When sacs containing PAM, 0.87 mM, and glucose, 10 mM, were placed in identical drug- and sugar-containing mediums, the inside (serosal) concentration of PAM fell by over half in 120 min, whereas that of glucose more than doubled. Mucosal hypertonicity depressed PAM efflux and glucose influx regardless of serosal osmolarity. Although azide and mucosal hypertonicity each depressed glucose uptake and oxygen consumption while accelerating net PAM influx, azide more effectively depressed glucose and oxygen uptake, whereas hypertonicity caused greater acceleration of PAM uptake. Hypertonicity did not affect PAM binding to intestinal tissue. Varying mucosal pH did not change PAM or glucose uptake. Thus, mucosal hypertonicity apparently enhances net mucosal-to-serosal transfer of PAM by blocking its active secretion from serosa to mucosa.     

Evidence for simultaneous bidirectional fluid flux across synovial lining in knee joints of anaesthetized rabbits.

A recent model predicted a local synovial circulation of extravascular fluid in which capillary filtrate entered a joint cavity and fluid simultaneously drained out of the cavity via intercellular pathways distant from capillaries. Here, solutions of albumin (10-250 g l-1) were infused continuously into the knee joint cavity in fifteen anaesthetized rabbits at 3-18 cmH2O pressure and steady-state net trans-synovial flow was recorded. The intra-articular fluid collected after 15-30 min was substantially diluted despite a sustained net efflux from the cavity. This supports the prediction that net efflux comprises an influx of low-protein plasma ultrafiltrate and a larger efflux of joint fluid.     

Magnesium net fluxes and distribution in rabbit myocardium in irreversible contracture

Distribution of magnesium (Mg) in heart muscle was studied by measuring fluxes of Mg and transmembrane potentials as a function of perfusate [Mg2+] after a massive increase in permeability of the sarcolemma was induced in the Langendorff prepared heart from the Nembutal-anesthetized rabbit. After onset of 0 mM [Ca2+] perfusion which produced excitation-contraction (E-C) uncoupling and mechanical arrest, action potentials recorded from subepicardial cells showed an increase in duration and decrease in amplitude, which progressed until no transmembrane potentials could be observed. Restoration of physiological salt solution perfusion after 15 min of [Ca2+]-free perfusion caused an irreversible contracture that was associated with 1) efflux of potassium (K) and myoglobin, 2) perfusate [Mg2+]-dependent flux of Mg, and 3) transmembrane potentials of 0 mV. The magnitude of net efflux of K and myoglobin during contracture was unaffected by perfusate [Mg2+]. During the first 2 min of contracture, net efflux of Mg (mumoles per gram wet muscle +/- SE) was 1.37 +/- 0.09 and 0.48 +/- 0.19 during 0 mM and 2.5 mM [Mg2+] perfusion, respectively; but a net influx of 0.56 +/- 0.23 occurred during 5 mM [Mg2+] perfusion. Total sarcoplasmic [Mg] may correspond to perfusate [Mg2+] of 3.6 mM, which was found by interpolation to prevent any net flux of Mg during contracture. 3.6 mM may, therefore, represent the upper limit of the intracellular free-ionized Mg concentration in rabbit heart.     

Effect of acetazolamide on sodium and chloride transport by in vitro rabbit ileum.

Acetazolamide (8 mM) aboishes active Cl absorption and inhibits but does not abolish active Na absorption by stripped, short-circuited rabbit ileum. These effects are not accompanied by significant changes in the transmural electrical potential difference or short-circuit current. Studies of the undirectional influxes of Na andCl indicate that acetazolamide inhibits the neutral, coupled NaCl influx process at the mucosal membranes. This action appears to explain the observed effect of acetazolamide on active, transepithelial Na and Cl transport. Acetazolamide did not significantly inhibit either spontaneous or theophylline-induced Cl secretion by this preparation, suggesting that the theophylline-induced secretion may not simply be due tothe unmasking of a preexisting efflux process when the neutral influx mechanism is inhibited by theophylline. Finally, inhibition of the neutral NaCl influx process by acetazolamide does not appear to be attributable to an inhibition of endogenous HCO3production or an elevation in intracellular cyclic-AMP levels. Instead, it appearstheat the effect of acetazolamide is due to a direct interaction with a membrane component involved in the coupled influx process.     

Development of the intestinal SGLT1 transporter in rats

Glucose absorption from the small intestine is largely mediated via the sodium-coupled glucose transporter (SGLT1). The goal of this study was to investigate the ontogenesis of the SGLT1, using the rat as an animal model at three stages of development: during lactation, at weaning, and at physiologic maturity. The techniques involved upper small intestinal perfusions with solutions containing 200 mM glucose and 50 mM NaCl, with or without 1 mM phloridzin (Phl), as an inhibitor of SGLT1. Molecular expression of the SGLT1 was also investigated via Western blot analysis from intestinal specimens of the three growth periods. Glucose absorption in weanling rats, in the absence of Phl, was several times higher than in sucklings and approximately double that of mature animals, and the effects of Phl were the greatest in weanlings. Furthermore, the physiologic data correlate to the molecular analysis of the SGLT1 which showed an increase in expression of the SGLT1 in both the weanlings and the adults compared to the sucklings. At all three stages of development Phl abolished Na absorption, and in sucklings there was a net outflow of Na. Due to the coupling between Na and water transport, net water absorption and the influx/efflux ratio, a more sensitive indicator of changes in unidirectional fluid movement, were similarly affected by Phl at the three stages of development. Net water absorption was highest in weanling animals. These findings are consistent with an early development of SGLT1 in rat small intestine and an apparent burst of activity at weaning. Less than complete maturity of other absorptive mechansims is occurring at this time.     

Urea degradation in sheep nourished by intragastric infusion: effects of level and nature of energy inputs

Four female sheep nourished wholly by infusions of volatile fatty acids (VFA), buffer and minerals to the rumen and casein to the abomasum were given in addition infusions of supplementary energy calculated to increase the energy input to the rumen by 30%. The design was a Latin square and the supplements given were (a) nil, (b) a standard VFA mixture similar to the basal infusion, (c) butyric acid alone and (d) glucose. Measurements were made of nitrogen retention and rumen fermentation characteristics and the kinetics of urea metabolism were measured over 24 h by means of a single injection of [14C]urea. An active microbial fermentation was established in the rumen in response to the infusion of glucose and estimates of microbial protein synthesis derived from urinary purine excretion agreed well with those calculated from stoichiometric principles. The presence of a microbial population in the rumen resulted in a decrease in urinary urea excretion and reductions in plasma urea concentration, urea pool size and rumen ammonia (NH3) concentration. Infusion of the mixed VFA or butyric acid supplements had no effect on these indices of urea metabolism. Measurements of urea irreversible loss rate showed high variability and the mean values did not differ significantly between the four treatments. Urea degradation in the gastrointestinal tract was also highly variable but increased, on average, by 2.4 g day-1 on the high-energy treatments. Examination of regression relationships between these variables also indicated a difference between the glucose treatment and the others in the metabolic fate of the NH3 derived from urea hydrolysis. It is concluded that urea degradation increased in response to additions of energy but did not differ according to the nature of the supplements supplied. In the glucose-supplemented group, the NH3 arising from degraded urea was incorporated into microbial protein and so removed from the urea-NH3 cycle; when additions of mixed VFA or butyric acid were given, the NH3 arising in hydrolysis appeared simply to be reabsorbed as NH3 and to contribute anew to urea formation.     

The kinetics of influx of calcium and strontium into rat intestine in vitro

1. The role of uptake across the brush border in the intestinal absorption of calcium has been studied by examining the kinetics of influx into slices of rat intestine in vitro. Both mucosal and serosal surfaces were exposed to the medium.2. The rate of influx was accurately defined by a two-component expression comprising a saturable (Michaelis-Menten) term and a second term linear with concentration. Influx across the mucosal surface of closed sacs was similar, and the saturable component for slice influx could be ascribed mainly to transport across the mucosal surface. The half-saturation constant for Ca was near 1 mM. This component was predominant at normal luminal concentrations of free Ca in the duodenum of young rats, but less so in jejunum and ileum and in older rats.3. The same kinetic expression applied to Sr influx, with a half-saturation constant of 2-3 mM, and possibly also to Ba with an even higher value.4. The saturable component of Ca influx was greatly reduced by 2,4: dinitrophenol (DNP); influx was also inhibited by iodoacetate, cyanide and at 0 degrees C. Inhibition commenced soon after exposure of the slices. A high concentration of DNP also caused an increase in the linear component of Ca influx.5. The kinetics of Ca influx across the mucosal surface agreed closely with the kinetics of steady-state absorption of Ca either across the whole mucosal epithelium in vivo or across the entire intestinal wall in vitro. This agreement supports the hypothesis that Ca entry across the brush border is the rate-limiting step in absorption; such a hypothesis would allow net Ca translocation while preserving a low intracellular concentration of ionic Ca in the mucosal epithelial cells.     

The effect of substances releasing intracellular calcium ions on sodium-dependent calcium efflux from guinea-pig auricles.

1. 45-Ca efflux and resting tension were measured in isolated guinea-pig auricles under conditions known to change the intracellular free Ca ion concentration. 2. In the presence of [Na]o, caffeine (2mM) increases 45-Ca efflux, but does not produce a contracture, while in the absence of [Na]o and [Ca]o caffeine causes a contracture without increasing 45-Ca efflux. Adrenaline (10-minus5-10-minus 4M) with or without theophylline (0-5-1-0mM) has no effect on either 45-Ca efflux or resting tension. 3. In the presence of caffeine the rate of net efflux of Ca depends on [Na]o-2. Caffeine contractures of muscles in Na-free solution relax upon the addition of [Na]o. Relaxation is correlated with the increase in net efflux of Ca. 4. Cyanide (2mM) produces a variable increase in 45-Ca efflux without a concomitant contracture in Na-containing solutions, but in Na, Ca-free solutions a large contracture occurs without significant increase in 45-Ca efflux. 5. A large increase in 45-Ca efflux and a contracture were observed with the 'Ca-ionophore' X 537 A. 6. Changes in membrane potential (K-depolarization) in hypertonic solutions have no significant effect on Na-dependent 45-Ca efflux, which is an agreement with an electroneutral 2:1 Na-Ca exchange. 7. Cyanide and X 537 A both cause a considerable release of Ca ions from isolated guinea-pig heart mitochondria, while caffeine has no effect. 8. The results suggest a powerful role of the Na-Ca exchange system in reducing the intracellular Ca concentration after Ca release from intracellular stores.     

Transport of potassium by the colon of normal and sodium-depleted rats

1. Ascending and descending segments of colon of normal and Nadepleted rats were perfused with solutions of differing KCl concentration. Net K flux, electrical p.d. and, in some experiments, unidirectional K fluxes were measured.2. Variation of luminal K concentration over the range 0-40 mM did not affect p.d. or K efflux rate.3. K secretion rate fell about 30% when Na-free choline chloride solution was perfused.4. Net flux was a linear function of the luminal K concentration, and fell as the latter increased. Na depletion increased K secretion rate and passive permeability of the mucosa to K. Adrenalectomy had the reverse effect. The luminal K concentration, associated with zero net K flux was much greater than expected if the colonic mucosa behaved passively with respect to K.5. Unidirectional fluxes determined when 5 mM-KCl was in the lumen showed that the ratio influx/efflux was much less than predicted by Ussing's flux ratio equation.6. It was concluded that K influx was due to simple diffusion and K efflux to diffusion and active transport, both processes being increased by Na depletion.     

Effects of acute hepatic encephalopathy and in vitro treatment with ammonia on glutamate oxidation in bulk-isolated astrocytes and mitochondria of the rat brain.

The metabolism of [1-14C] glutamate to 14CO2 and the glutamate dehydrogenase (GLDH) activity towards alpha-ketoglutarate (alpha-KG) formation were measured in bulk isolated astrocytes derived from control rats and rats with acute hepatic encephalopathy (HE) induced with thioacetamide. In addition, the effects of in vitro treatment of control and HE astrocytes and non-synaptic mitochondria with toxic (3mM) NH4Cl concentration were followed. [1-14C] glutamate oxidation measured as a whole was identical in control and HE astrocytes and was inhibited by ammonia to the same degree in either fraction. In the presence of a glutamate transamination inhibitor--3mM aminooxyacetic acid (AOA), when only the GLDH-mediated part (25% of total) of the glutamate oxidation remained active, the inhibitory effect of ammonia treatment was much more pronounced in HE astrocytes than in control astrocytes. The ability of non-synaptic mitochondria to utilize glutamate to CO2 was not changed in presence of 3mM NH4Cl, whereas a substantial decrease of CO2 production (about 80%) in both the control and HE preparations was observed in the presence of 3mM AOA. GLDH activity was not at all affected by either of the experimental conditions, both in astrocytes and purified non-synaptic mitochondria. Thus, the inhibition of glutamate oxidation in astrocytes by ammonia and the compounded inhibitory effect of HE, ammonia and AOA appeared to be located beyond the glutamate dehydrogenation step within the tricarboxylic acid cycle.     

The effect of the internal sodium concentration on calcium fluxes in isolated guinea-pig auricles

1. Calcium efflux from guinea-pig auricles followed saturation kinetics when [Ca](o) and [Na](o) were changed while the ratio [Ca](o)/[Na](o) (2) was kept constant. The Michaelis constant, K(m) (Ca+Na) = 40 mM, suggests that a hypothetical carrier system, responsible for sodium-calcium exchange, is far from saturation with the inside concentrations of these ions.2. [Na](i) was altered in the auricles between 12.5 and 60 mM/kg fibre water while total cellular calcium concentration ([Ca](t)) at the beginning of the influx period was not significantly different in the various groups of preparations.3. (45)Ca influx increased appreciably with increasing [Na](i). (45)Ca influx from sodium-poor solution corresponded to an almost equal increase in [Ca](t), while [Ca](t) did not change much in preparations loaded with (45)Ca in Tyrode solution. When the sodium-activated fraction of calcium influx was plotted against [Na](i) (2) the resulting curve indicated saturation with K(m) (Na) = 3500 (mM [Na](i))(2) and maximal influx rate, J(i, max) (Ca') = 1.35 mM/kg wet weight x 10 min.4. When the preparations were re-equilibrated for various times in normal Tyrode solution after [Na](i) had been increased, both the sodium-activated component of calcium influx and [Na](i) (2) decreased with approximately the same rate constants.5. Calcium efflux from auricles with high [Na](i) was increased when it was measured in Tyrode solution while the efflux in sodium-poor solution was inhibited.6. Auricles with increased [Na](i) showed a positive inotropic contractile response.7. The main conclusion reached by these experiments is that calcium influx is affected by [Na](i) in a way which is compatible with a carrier-mediated sodium-calcium exchange system.     

Micropuncture studies of the osmoregulation in the nauplius of Artemia salina

The osmoregulation of the nauplius of the brine shrimp, Artemia salina, was investigated using micropuncture and microanalytical techniques. The naupliar body fluid, hemolymph was hyposmotic to and had lower Na concentrations than the suspending medium for the range of medium salinities from 80 to 4,900 mM NaCl. In medium containing 20 mM NaCl, the hemolymph was hyperosmotic to the medium, with osmolarity of 101 +/- 8 mosmol/1 and with [Na] of 49 +/- 11 meq/1. Whereas the maximal observed NaCl concentration gradient between hemolymph and medium was 4,785 mM, during the incubation of nauplii in artificial seawater (osmolarity: 932 mosmol/1; and [Na]: 502 meq/1) the osmolarity and [Na] of the naupliar hemolymph were 161 +/- SD 16 mosmol/1 and 86 +/- 14 meq/1, respectively. The influx and efflux of Na between medium and hemolymph were measured using 22Na. The fluxes of this ion were temperature dependent. The main site of efflux of 22Na was the neck organ as was shown by experiments of differential recovery of 22Na introduced in the hemolymph. These studies demonstrate that the nauplius of A. salina has the ability to osmoregulate not only against high environmental salinities but also against low salinities approaching those of freshwater.     

Thallium and the sodium pump in human red cells

1. Thallium (Tl) inhibits the ouabain-sensitive K influx in human red cells in high-Na medium. At 1 mM external K concentration [K(o)], the ouabain-sensitive K influx decreases steadily with increasing Tl concentration, up to 0.9 mM outside; at 0.17 mM-K(o), however, Tl stimulates the ouabain-sensitive K influx below 0.1 mM-Tl(o) and inhibits it at higher concentrations.2. In a K-free medium in which all except 5 mM-Na is replaced by choline, and into which red cells show zero control ouabain-sensitive Na efflux, Tl is able to support ouabain-sensitive Na efflux up to 2.1 m-mole/l. cells.hr following a sigmoid activation curve which is half-maximal between 0.03 and 0.05 mM-Tl(o) and that follows two-site kinetics up to 0.1 mM-Tl(o). Beyond 0.15 mM-Tl(o), the Tl-activated ouabain-sensitive Na efflux attained is inhibited slightly.3. When the ouabain-sensitive Na efflux is measured at 5 mM-Na(o) and 5 mM-K(o), increasing concentrations of Tl have little effect on it, 0.9 mM-Tl(o) inhibiting by some 14%; in similar conditions, the ouabain-sensitive K influx is inhibited by about 40%.4. The dependence of ouabain-sensitive K influx on external K concentration at 5 mM-Na(o), which follows a slightly sigmoid curve in the absence of Tl, changes to hyperbolic at 0.06 mM-Tl(o) at the same time that ouabain-sensitive K influx is inhibited. The fitted V(max) values for ouabain-sensitive K influx are the same in the presence and in the absence of 0.06 mM-Tl(o).5. In high-Na cells, loaded by nystatin treatment, the ouabain-sensitive K influx measured at 0.2 mM-Na(o) follows a hyperbolic curve between 0.05 and 0.4 mM-K(o), and is inhibited by Tl in a strictly competitive fashion.6. The effects of Tl on ouabain-sensitive Na efflux and ouabain-sensitive K influx are interpreted in terms of a high-affinity substitution for K at the external K sites of the Na pump and suggest that in human red cells Tl can be actively transported inwards in exchange for internal Na.7. Thallium can inhibit about 25% of the ouabain-insensitive Na efflux into 5 mM-Na(o) and part of this inhibition occurs with a high Tl-affinity; the ouabain-insensitive K influx is inhibited by Tl both in high-Na and in 5 mM-Na medium, but with a different concentration dependence than the ouabain-insensitive Na efflux.