Erythrocyte sodium content and transport in borderline and mild hypertension

Summary Sodium content and transport of red blood cells were examined in 98 male blood donors. Regarding their blood pressure they were classified into the following groups: (a) 57 normotensives, (RR<140/90 mm Hg); (b) 24 borderline hypertensives (140/90RR<160/95 mm Hg); and (c) 17 hypertensives (RR>160/100 mm Hg). Compared with the normotensives the borderline hypertensives have significantly reduced red cell sodium content. The ouabain-resistant net Na⁺ uptake and the relative Na⁺ uptake, as a measure of the Na⁺/K⁺ pump, were significantly increased. With rising blood pressures the measured values turn to normal, so that no difference exists between the normotensive and hypertensive groups. It is supposed that in the initial or even prehypertensive state a considerable enhancement of the pump activity occurs, simultaneously accompanied by less marked increases in sodium influx, leading to a reduced intracellular sodium content. In the course of hypertension, possibly caused by the formation of a pump inhibitor, the sodium content of red cells turns to normal or supernormal values.Abbreviations BMI body mass index - BHT borderline hypertensive - Ca _ion ²⁺ ionized plasma calcium - HT hypertensive - k relative OR net Na⁺ uptake - [Na⁺]_i, [K⁺]_i intracellular sodium and potassium content in RBCs - NT normotensive - OR ouabain-resistant - RBCs red blood cells - Na OR net Na⁺ uptake     

Evidence for coupled sodium/hydrogen exchange in the rat superficial proximal convoluted tubule

Recent in vitro studies from the rat and rabbit have suggested a tightly coupled sodium/hydrogen ion exchanger on the luminal membrane of proximal tubules. The steep sodium gradient from the lumen to cell supplies indirect energy for hydrogen ions to be pumped from the cell to the lumen. However, a proton translocating pump has been demonstrated in other epithelia, which is independent of sodium transport and directly driven by ATP. To examine the role that sodium might play in the process of acidification, rat proximal convoluted tubules and their surrounding peritubular capillaries were perfused in vivo with artificial ultrafiltrate-like perfusion solutions. Total CO₂ absorption was measured by microcalorimetry during alterations in sodium transport by replacement of the sodium with an impermeant cation, choline, or by inhibition of the (Na⁺+K⁺)-ATPase by removing potassium from both perfusion solutions. Under control conditions the absolute rate of total CO₂ absorption was 140 pmol/mm·min. In the choline substitution and potassium removal experiments, absolute total CO₂ absorption fell to 23 and 28 pmol/mm·min, respectively. The data suggest that: 1) in the rat superficial proximal convoluted tubule approximately 80% of the bicarbonate absorption is tightly coupled to sodium transport; 2) this process is driven indirectly by the (Na⁺+K⁺)-ATPase system; and 3) the residual 20% of acidification appears to be mediated by another mechanism or may be a consequence of technical liminations.     

Abnormal erythrocyte sodium leak in a subset of essential hypertensive patients

Summary We measured the ouabain- and bumetanide-resistant Na⁺ efflux in Mg²⁺-sucrose medium (passive Na⁺ leak) in erythrocytes from 30 normotensive controls and 72 essential hypertensive patients. The mean values (±SEM) of the rate constant of Na⁺ leak (k_pNa) were not significantly different between normotensives and hypertensives. Nevertheless, using the 95% confidence limits of the k_pNa (in 10^–3.h^–1) in the normotensive group as a cut-off point, 7 (9.7%) essential hypertensives exhibited increased values (58.96±10.12) when compared with the other 65 patients (23.86±0.74). revealing increased passive Na⁺ permeability in the former (leak + hypertensives). Na⁺ fluxes depending on the Na⁺-K⁺ pump, outward Na⁺-K⁺ cotransport, and Na⁺-Li⁺ countertransport were also measured in fresh erythrocytes from the same 72 patients. Three of them (4.2%) exhibited decreased values of ouabain-sensitive Na⁺ efflux and 6 (8.3%) of bumetanide-sensitive Na⁺ efflux, while 8 patients (11.1%) showed increased values of Li⁺-stimulated Na⁺ efflux and, finally, 48 patients (59.7%) did not present any evident abnormality in these Na⁺ transport systems. No differences were observed between leak + hypertensives and the remaining 65 patients when both basal erythrocyte Na⁺ content and clinical parameters of hypertension were compared. However, Na⁺ efflux depending on the outward Na⁺-K⁺ cotransport was significantly higher in the leak + hypertensive subset (299.43±43.18 vs 181.52±10.76 µmol.(l cells.h)^–1;P=0.0078), suggesting a compensatory phenomenon. Enhancement of Na⁺ permeability detected in 3% to 16% of essential hypertensives may be implicated in the pathogenesis of primary hypertension.Abbreviations ATPase adenosine triphosphatase - Dcat difference between the external Na⁺ concentration after incubation at 37° C and at zero time - k_pNa rate constant of passive Na⁺ leak - Leak + hypertensive essential hypertensive patient with abnormally high erythrocyte Na⁺ leak - MOPS 4-morpholinopropanesulfonic acid - OBR ouabain- and bumetanide-resistant - PRA plasma renin activity - sPRA plasma renin activity stimulated after furosemide infusion - SEM standard error of the mean Supported in part by Grant 87/1078 of the Fondo de Investigaciones Sanitarias de la Seguridad Social and Grant PA85/0168 of the Comisión Asesora de Investigación Científica y Técnica     

Na+-Li+ countertransport and electrolyte composition in erythrocytes of patients with essential hypertension before and after antihypertensive treatment

Summary The effect of antihypertensive treatment with 6–12 mg of the loop diuretic piretanide over 12 weeks on Na⁺-Li⁺ countertransport, and on extra- and intracellular electrolyte composition was studied in 10 previously untreated patients with essential hypertension. These data were compared with 10 sex- and age-matched controls. Blood pressure fell from 180±18.3/110.5±9.8 to 154.7±9.7/92.5±10.9 mmHg during treatment. Na⁺-Li⁺ countertransport was significantly higher in hypertensives (0.36±0.13 mmol×l _rbc ^–1 ×h^–1) compared with controls (0.25±0.05;P<0.05).Na⁺-Li⁺ countertransport and intracellular electrolyte composition remained unchanged whereas the extracellular potassium concentration fell from 4.28±0.51 to 3.98±0.36 mmol/l (P<0.05). No increase in the intracellular Na⁺ content as compared with normotensive controls was found. It is concluded that the intracellular Na⁺ concentration is not a marker for essential hypertension. The Na⁺-Li⁺ countertransport does not seem to be directly related to elevated blood pressure but seems to be a general marker for diseases associated with an increased risk for the development of hypertension.Supported by the DFG (Er 65/4-4)     

Sodium pump quantity and turnover in rabbit descending colon at different rates of sodium absorption

³H-Ouabain binding to isolated epithelia and basolateral membrane vesicles of Na⁺-transporting epithelial cells of rabbit descending colon was determined to quantify the number of operative Na⁺-pump sites at different rates of transcellular Na⁺ transport which was varied over a wide range by chronic dietary Na⁺ restriction or Na⁺ loading. Both in intact epithelia and in basolateral membrane vesicles the maximal number of specific ouabain binding sites was higher in preparations from animals transporting Na⁺ at high rates than in preparations from animals transporting Na⁺ at low rates. The affinity of ouabain to its binding site and the association and dissociation rate constants were not dependent on the rate of Na⁺ transport. In intact epithelia the Na⁺ turnover rate per pump unit was twice as high in tissues with high Na⁺ transport than in tissues with low Na⁺ transport. In basolateral membrane vesicles the Na⁺ turnover rate was considerably higher than in intact epithelia and there was no difference in turnover rate between vesicle preparations obtained from tissues transporting Na⁺ at high or low rates. Hence, factors within the intact cell appear to control the turnover rate of the Na⁺-pump.     

Dependence of the electrogenic pump current of Xenopus oocytes on external potassium

The membrane potential of Xenopus oocytes showed a variable response to an increase of the K⁺ concentration in the bathing solution, [K⁺]_e, from 2.5 mM to 20 mM. In 54% of the cases (n=52) the cells hyperpolarized (by max. 70 mV). In the presence of 10^–5 M ouabain, all cells depolarized suggesting that the hyperpolarization was caused by an electrogenic Na⁺/K⁺ pump. In cells stored overnight in a Na⁺-free solution the transition from 2.5 to 20 mM [K⁺]_e always caused depolarization indicating that the stimulation of the pump requires high internal sodium, [Na⁺]_i. Cells stored overnight in a Na⁺-rich solution had a [Na⁺]_i of 30.7±7 mM, i.e. the Na⁺/K⁺ pump was saturated with sodium (Lafaire and Schwarz 1986). With 9 such cells we determined the K⁺ activation of the Na⁺/K⁺ pump. The activation follows Hill kinetics with I_max=90.5 nA, K_s=2.3 mM, and n=1.68.     

Cholinergic and GABAergic innervation regulate activity of electrogenic ionic pump in earthworm somatic muscle cells

Carbacholine reduced, while baclofen and norepinephrine increased resting membrane potential in earthworm somatic muscle cells. In the presence of carbacholine, neither norepinephrine, nor baclofen hyperpolarized the membrane. Ouabain decreased resting potential and abolished the effects of carbacholine, norepinephrine, and baclofen on membrane potential. It was hypothesized that carbacholine directly inhibited the ouabain-sensitive component of Na⁺/K⁺ pump and abolished the activating effect of norepinephrine and baclofen. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 144, No. 8, pp. 128–130, August, 2007     

Submandibular salivary secretion in the cat and associated potassium movements: Dependence on temperature and perfusate flow rate

Cat submandibular glands were perfused with Locke solution in a thermostated chamber and intermittently stimulated with 10^–5 M acetylcholine (ACh). In one series of experiments the perfusion pressure was varied within the range 90–60 mm Hg, and secretory flow rate, active K⁺-reuptake, passive K⁺-release, and resting and ACh-induced venous flow rates were measured. The ACh-induced secretory flow rate and the maximal K⁺-fluxes were related to the simultaneous ACh-induced venous flow rates. A proportionality was found between the maximal rate of ACh-induced K⁺-release and ACh-induced venous flow rates below 8 ml/min, while at higher flow rates the K⁺-release leveled off. The maximal rate of the post-stimulatory K⁺-reuptake increased proportionally to the ACh-induced perfusate flow rate throughout the range studied. The secretory flow rate was much less affected by changes in ACh-induced perfusate flow rate. In another series of experiments the gland temperature was varied within the range 12–37°C, and the same parameters were measured. All parameters decreased with cooling being reduced to 50% of their 37°C values at: 24°C for secretion, 19°C for K⁺-reuptake, and 14°C for K⁺-release. It is concluded: that 1) the rate of ACh-induced K⁺-release is limited by the ACh-induced perfusate flow rate (within the physiological range), 2) the capacity of the K⁺-reuptake mechanism is at least one order of magnitude larger than the maximal rate of K⁺-reuptake in vivo, 3) the marked temperature sensitivity of the secretory flow rate reflects the high complexity of the mechanisms involved.     

Effect of sodium concentration and of atropine on the contractile response of the guinea-pig ileum to potassium ions

The possibility that the guinea-pig ileum's contractile response to K⁺-rich solutions is partly mediated by acetylcholine release from the intramural nervous tissue was examined by studying the inhibition of that response by atropine at different values of [Na⁺]₀. In a medium in which the NaCl was replaced by iso-osmotic glucose, the response to high [K⁺]₀ was not greatly affected, while the responses to acetylcholine and to other agonists were significantly reduced. In control medium (149 mM Na⁺), atropine (10^–7 M) partly inhibited the responses to K⁺-rich solutions and to agonists such as histamine, 5-hydroxytryptamine and bradykinin. When [Na⁺]₀ was reduced to 12 mM, by iso-osmotic substitution of glucose for NaCl, the response to high K⁺ was no longer inhibited by atropine, which still partly inhibited the contractions elicited by the three agonists and totally blocked the response to acetylcholine. It is proposed that atropine's inhibition of the response to high K⁺ and to agonists is not due to its specific anti-muscarinic effect, but to an unspecific action, which in the case of the agonists is independent of [Na⁺]₀. In addition, the inhibition of the response to high K⁺ would result from a different Na⁺-dependent mechanism, possibly involving the stimulation of the Na-K pump by atropine. This is supported by the observation that this drug partly relaxed ileum preparations that were contracted by ouabain.     

Erythrocyte cations and Na+,K+-ATPase pump activity in athletes and sedentary subjects

Summary The chronic effect of training on intraerythrocyte cationic concentrations and on red cell Na⁺,K⁺-ATPase pump activity was studied by comparing well-trained athletes with sedentary subjects at rest. Also the acute effect of a 50-min cross-country run on these erythrocyte measurements was studied in the athletes. At rest the intraerythrocyte potassium concentration was increased (P<0.01) in the athletes compared to that of the control subjects. The intraerythrocyte concentrations of sodium and magnesium and red cell Na⁺, K⁺-ATPase pump activity were, however, similar in the trained and the untrained subjects.As compared with the resting condition, the intraerythrocyte potassium concentration was decreased (P<0.05) after exercise in the athletes, and this was accompanied by a minor increase in the intraerythrocyte sodium concentration. Red cell Na⁺,K⁺-ATPase pump activity was slightly increased (P<0.05) after exercise.     

Effects of aldosterone on intralymphocytic sodium and potassium in patients with essential hypertension

Summary In vitro binding of aldosterone to mineralocorticoid receptors on human mononuclear leukocytes (HML) and its effects on the intracellular sodium and potassium concentrations of HML have already been described. In the present paper this easily accessible human cell model was investigated in 13 patients with essential hypertension. In only four patients sodium in HML without incubation was elevated compared with the range for normal persons. A decrease of intracellular sodium or potassium occurred during incubation without aldosterone (P<0.02). The addition of 1.4 nM aldosterone did not prevent this loss of electrolytes as observed in normal persons. Plasma renin activity and aldosterone were not correlated with the electrolyte response and were within the normal limits. The number of mineralocorticoid receptors/cell were within or close to the normal range (n=9). The independence of intracellular electrolytes from aldosterone despite a normal number of mineralocorticoid receptors may reflect an impairment of the mineralocorticoid effector mechanism in the HML of patients with essential hypertension.Abbreviations HML human mononuclear leukocytes - HEPES N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid - Tris Tris(hydroxymethyl)aminomethane     

SIK1/SOS2 networks: decoding sodium signals via calcium-responsive protein kinase pathways

Changes in cellular ion levels can modulate distinct signaling networks aimed at correcting major disruptions in ion balances that might otherwise threaten cell growth and development. Salt-inducible kinase 1 (SIK1) and salt overly sensitive 2 (SOS2) are key protein kinases within such networks in mammalian and plant cells, respectively. In animals, SIK1 expression and activity are regulated in response to the salt content of the diet, and in plants SOS2 activity is controlled by the salinity of the soil. The specific ionic stress (elevated intracellular sodium) is followed by changes in intracellular calcium; the calcium signals are sensed by calcium-binding proteins and lead to activation of SIK1 or SOS2. These kinases target major plasma membrane transporters such as the Na⁺,K⁺-ATPase in mammalian cells, and Na⁺/H⁺ exchangers in the plasma membrane and membranes of intracellular vacuoles of plant cells. Activation of these networks prevents abnormal increases in intracellular sodium concentration.     

Influence of Mitragyna Ciliata (MYTA) on the Microsomal Activity of ATPase Na+/K+ Dependent Extract on a Rabbit Heart

Mitragyna ciliata (MYTA) (Rubiaceae) inhibits plasmodia activity. MYTA induces a cardiotonicity of the digitalic type on rat''s isolated heart. In this work we studied the effect of MYTA on microsomal Na⁺/K⁺ dependant ATPase (Na⁺, K⁺ ATPase) extracted from the heart of a rabbit since digitalics inhibit Na⁺, K⁺ ATPase. Our results revealed that the Na⁺/K⁺ ATPase has an optimum pH of 7.4 and temperature of 37°C respectively. There is a linear relationship between the organic phosphate formed and the incubation time over 25 mins incubation period. The ATP hydrolysis rate in the presence of MYTA was 0.775 µM/min. LINEWEAVER and BURK plots showed that MYTA did not alter K_M (1.31 mM) but decreased V_MAX. This study shows that MYTA exerts a non-competitive inhibition on the microsomal Na⁺/K⁺ ATPase extracted from rabbit heart with a Ci₅₀ of 48 µg / ml. We conclude that the mechanism of action of MYTA is linked to the inhibition of the Na⁺/K⁺ ATPase like cardiotonics of the digitalic type.     

(Na+K+)-activated ATPase in human cornea

Summary Distribution and principal characteristics of (Na⁺K⁺)-activated ATPase in human cornea were investigated.(Na⁺K⁺)-ATPase was present in both epithelium and endothelium, whereas the corneal stroma did not exhibit significant enzyme activity.In homogenates specific activity of the (Na⁺K⁺)-ATPase was 2.3-fold higher in endothelium than in epithelium. Calculation of total enzyme activity revealed a 6.1-fold higher content of (Na⁺K⁺)-ATPase in the epithelium.In the epithelium a 7-fold enrichment of (Na⁺K⁺)-ATPase compared to the homogenate was obtained in the 150–1500×g _av fraction. Maximum enrichment in the endothelium was 3.5-fold and was achieved in the 1500–2500×g _av fraction. Both fractions showed, however, the same specific activity.The pH-optimum of (Na⁺K⁺)-ATPase in the 150–1500×g _av fraction ranged from 8.0–8.2 in both epithelium and endothelium.In the epithelial 150–1500×g _av fraction the apparentK _m-values were 4.0 mM for Na⁺, 2.8 mM for K⁺ and 0.12 mM for Mg²⁺ · ATP in equimolar concentrations.The inhibition constant of epithelial (Na⁺K⁺)-ATPase for ouabain was determined asK _i=3.3×10^–7 M.The present data support the view that control of corneal hydration in man is a function of both endothelium and epithelium.     

Na+/K+ pump and endothelial cell survival: [Na+]i/[K+]i-independent necrosis triggered by ouabain, and protection against apoptosis mediated by elevation of [Na+]i

Recent studies have demonstrated the tissue-specific effect of Na⁺/K⁺ pump inhibition by ouabain and other cardiac glycosides on cell viability. The vascular endothelium is an initial target of cardiac glycosides employed for the management of congestive heart failure as well as circulating endogenous ouabain-like substances (EOLS), the production of which is augmented in volume-expanded hypertension. This study examined the role of the Na⁺/K⁺ pump in the survival of cultured porcine aortic endothelial cells (PAEC). Complete Na⁺/K⁺ pump inhibition with ouabain led to PAEC death, indicated by cell detachment and decreased staining with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). Based on cell swelling and resistance to benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone (z-VAD.fmk) a pan-caspase inhibitor, this type of cell death was classified as necrosis. In contrast to ouabain, Na⁺/K⁺ pump inhibition in K⁺-free medium did not affect PAEC viability and sharply attenuated apoptosis triggered by ³H decay-induced DNA damage. Necrosis evoked by ouabain was preserved after dissipation of the transmembrane gradient of K⁺ and Na⁺, whereas dissipation of the Na⁺ gradient abolished the antiapoptotic action of K⁺-free medium. Comparative analysis of these results and modulation of intracellular Na⁺ and K⁺ content by the above-listed stimuli showed that interaction of ouabain with Na⁺/K⁺-ATPase triggered necrosis independently of inhibition of Na⁺/K⁺ pump-mediated ion fluxes and inversion of the [Na⁺]_i/[K⁺]_i ratio, whereas protection against apoptosis under Na⁺/K⁺ pump inhibition in K⁺-depleted medium was mediated by [Na⁺]_i elevation. The role of Na⁺/K⁺ pump-mediated regulation of endothelial cell survival and vascular remodelling seen in hypertension should be investigated further in context of EOLS and chronic treatment with digitalis.     

Transcellular sodium transport and basolateral rubidium uptake in the isolated perfused cortical collecting duct

The relation between transcellular Na⁺ absorption, intracellular Na⁺ concentration and Na⁺/K⁺-ATPase activity (the last estimated by the rubidium uptake across the basolateral cell membrane) was examined in the different cell types of the rabbit cortical collecting duct (CCD). Experiments were performed on isolated perfused CCD in which Na⁺ absorption was varied by perfusing the tubule with solutions containing different Na⁺ concentrations (nominally Na⁺-free, 30 mM and 144 mM). Experiments were terminated by shock-freezing the tubules during perfusion. Precisely 30 s before shock-freezing, the K⁺ in the bathing solution was exchanged for Rb⁺. Intracellular element concentrations, including Rb⁺, were determined in freeze-dried cryosections of the tubules using energy-dispersive X-ray analysis. Increasing Na⁺ concentration in the perfusion solution caused significant rises in intracellular Na⁺ concentration and Rb⁺ uptake of principal cells. Principal cell Na⁺ and Rb⁺ concentrations were 7.8±0.9 and 7.0±0.8 mmol/kg wet weight respectively, when the perfusion solution was Na⁺-free, 10.1±0.7 and 11.6±0.6 mmol/kg wet weight with 30 mM Na⁺ in the perfusion solution, and 14.5±1.5 and 14.9 ±0.9 mmol/kg wet weight with 144 mM Na⁺ in the perfusion solution. In contrast, a comparable relationship between lumen Na⁺ concentration, intracellular Na⁺ concentration and basolateral Rb⁺ uptake was not seen in intercalated cells. These results support the notion that principal, but not intercalated, cells are involved in transepithelial Na⁺ absorption. In addition, the data demonstrate that apical Na⁺ entry and basolateral Na⁺/K⁺-AT-Pase activity are closely coupled in principal cells of the rabbit CCD. A rise in lumen Na⁺ concentration leads to increased Na⁺ entry and augmented intracellular Na⁺ concentration, which then secondarily stimulates active basolateral Na⁺/K⁺(Rb⁺) exchange.     

Membrane potential measurements of transitional cells from the crista ampullaris of the Gerbil

Transitional cells of the crista ampullaris were impaled with microelectrodes in order to record the membrane potential (PD) and to investigate membrane properties. In control solution the PD was –87±1 mV (n=103). This value is not significantly different from –83±2 mV (n=24) measured in Cl^– free solution. [Cl^–] steps from 150 to 15 mmol/l (n=24) depolarized the membrane by about 2 mV, indicating a minor Cl^– conductance. The transference number for K⁺ was 0.75±0.01 (n=79) obtained from the PD responses to K⁺ steps from 3.6 to 25 mmol/l. The cell membrane depolarized and the amplitude of PD responses to [K⁺] steps was reduced by Ba²⁺ (2·10^–6 to 10^–3 mol/l), quinidine (10^–3 mol/l), quinine (10^–3 mol/l), Rb⁺ (20 mmol/l), Cs⁺ (20 mmol/l), NH₄ ⁺ (20 mmol/l) and Tl⁺ (0.5 mmol/l), whereas tetraethylammonium (TEA, 20 mmol/l) had no effect. The dose-response curve for Ba²⁺ in the presence of 3.6 mmol/l K⁺ was shifted to the right by approximately three decades in the presence of 25 mmol/l K⁺ and by a factor of about 4 in the presence of 135 mmol/l gluconate as a substitute for Cl^–. Transitional cells were depolarized by ouabain, suggesting the presence of (Na⁺+K⁺-ATPase.This work was supported by grants from the Deafness Research Foundation to PhW and the National Institute of Health (NS 19490) to DCM     

The packing of acetylcholine into quanta at the frog neuromuscular junction is inhibited by increases in intracellular sodium

Pretreatment with hypertonic solutions, insulin, or adrenaline increases the size of quanta at the frog neuromuscular junction, as determined by measurements of miniature end plate potentials or currents (Van der Kloot and Van der Kloot 1985, 1986). The increase in quantal size apparently is due to an increase in acetylcholine (ACh) content of individual quanta. These treatments, therefore, can be used to study the packaging of ACh. Previously, I reported that increases are blocked by an inhibitor of active ACh uptake into vesicles (Van der Kloot 1986b, 1987b). The present study shows that the increases in quantal size were antagonized by inhibiting the Na⁺–K⁺ exchange pump with 100 M ouabain, 10 M dihydroouabain, or K⁺-free solutions. The increases in quantal size were also antagonized by 10 M monensin, a Na⁺ ionophore, or by 5 M aconitine, which opens Na⁺ channels at normal resting potentials. Apparently a rise in intracellular [Na⁺] inhibits the addition of ACh to quanta. The mechanism by which a rise in intracellular Na⁺ inhibits ACh packing is unknown, but apparently it is not due to inhibition of choline reuptake into the terminals.Also consistent with the above hypothesis is that the increase in quantal size following depolarization for 2 h in elevated [K⁺]_out was substantially enhanced when tetrodotoxin (TTX) was present, suggesting that in the absence of TTX there is a rise in [Na⁺]_in that antagonizes the incorporation of additional ACh into the quanta.     

Effect of high NaCl intake on Na+ and K+ transport in the rabbit distal convoluted tubule

Morphological studies have demonstrated that a chronic increase in distal Na⁺ delivery causes hypertrophy of the distal convoluted tubule (DCT). To examine whether high NaCl-intake also causes functional changes in the well defined DCT, we measured transmural voltage (V _T), lumen-to-bath Na⁺ flux (J _Na(LB)), and net K⁺ secretion (J _K(net)) in DCTs obtained from control rabbits and those on high NaCl-intake diets. The lumen negativeV _T was significantly greater in the high NaCl group than in the control group. The net K⁺ secretion (pmol mm^–1 min^–1) was greater in the high NaCl-intake group (54.1±13.0 vs 14.7±5.6). The K⁺ permeabïlities in both luminal and basolateral DCT membranes, as assessed by the K⁺-induced transepithelial voltage deflection inhibitable with Ba²⁺, were increased in the experimental group. The lumen-to-bath²²Na flux (pmol mm^–1 min^–1) was also greater in the experimental group (726±119 vs 396±65). TheV _T component inhibitable with amiloride was also elevated in the high NaCl-intake group. Furthermore, Na⁺–K⁺-ATPase activity of the DCT was higher in the experimental than in the control group. We conclude that high NaCl intake increases both Na⁺ reabsorption and K⁺ secretion by the DCT. This phenomenon is associated with an increased Na⁺–K⁺-ATPase activity along with increased Na⁺ and K⁺ permeabilities of the luminal membrane, and an increase in the K⁺ permeability of the basolateral membrane. Cellular mechanisms underlying these functional changes remain to be established.