K+ channel KVLQT1 located in the basolateral membrane of distal colonic epithelium is not essential for activating Cl- secretion

The cellular mechanism for Cl^– and K⁺ secretion in the colonic epithelium requires K⁺ channels in the basolateral and apical membranes. Colonic mucosa from guinea pig and rat were fixed, sectioned, and then probed with antibodies to the K⁺ channel proteins K_VLQT1 (Kcnq1) and minK-related peptide 2 (MiRP2, Kcne3). Immunofluorescence labeling for Kcnq1 was most prominent in the lateral membrane of crypt cells in rat colon. The guinea pig distal colon had distinct lateral membrane immunoreactivity for Kcnq1 in crypt and surface cells. In addition, Kcne3, an auxiliary subunit for Kcnq1, was detected in the lateral membrane of crypt and surface cells in guinea pig distal colon. Transepithelial short-circuit current (I_sc) and transepithelial conductance (G_t) were measured for colonic mucosa during secretory activation by epinephrine (EPI), prostaglandin E₂ (PGE₂), and carbachol (CCh). HMR1556 (10 µM), an inhibitor of Kcnq1 channels (Gerlach U, Brendel J, Lang HJ, Paulus EF, Weidmann K, Brüggemann A, Busch A, Suessbrich H, Bleich M, and Greger R. J Med Chem 44: 3831–3837, 2001), partially (50%) inhibited Cl^– secretory I_sc and G_t activated by PGE₂ and CCh in rat colon with an IC₅₀ of 55 nM, but in guinea pig distal colon Cl^– secretory I_sc and G_t were unaltered. EPI-activated K⁺-secretory I_sc and G_t also were essentially unaltered by HMR1556 in both rat and guinea pig colon. Although immunofluorescence labeling with a Kcnq1 antibody supported the basolateral membrane presence in colonic epithelium of the guinea pig as well as the rat, the Kcnq1 K⁺ channel is not an essential component for producing Cl^– secretion. Other K⁺ channels present in the basolateral membrane presumably must also contribute directly to the K⁺ conductance necessary for K⁺ exit during activation of Cl^– secretion in the colonic mucosa. HMR1556; K⁺ secretion; epinephrine; prostaglandin E₂; cholinergic     

Cl- secretory effects of EBIO in the rabbit conjunctival epithelium

Experiments were conducted to determine whether the Cl^– secretagogue, 1-ethyl-2-benzimidazolinone (EBIO), stimulates Cl^– transport in the rabbit conjunctival epithelium. For this study, epithelia were isolated in an Ussing-type chamber under short-circuit conditions. The effects of EBIO on the short-circuit current (I_sc) and transepithelial resistance (R_t) were measured under physiological conditions, as well as in experiments with altered electrolyte concentrations. Addition of 0.5 mM EBIO to the apical bath stimulated the control I_sc by 64% and reduced R_t by 21% (P < 0.05; paired data). Under Cl^–-free conditions, I_sc stimulation using EBIO was markedly attenuated. In the presence of an apical-to-basolateral K⁺ gradient and permeabilization of the apical membrane, the majority of the I_sc reflected the transcellular movement of K⁺ via basolateral K⁺ channels. Under these conditions, EBIO in combination with A23187 elicited nearly instantaneous 60–90% increases in I_sc that were sensitive to the calmodulin antagonist calmidazolium and the K⁺ channel blocker tetraethyl ammonium. In the presence of an apical-to-basolateral Cl^– gradient and nystatin permeabilization of the basolateral aspect, EBIO increased the Cl^–-dependent I_sc, an effect prevented by the channel blocker glibenclamide (0.3 mM). The latter compound also was used to determine the proportion of EBIO-evoked unidirectional ³⁶Cl^– fluxes in the presence of the Cl^– gradient that traversed the epithelium transcellularly. Overall, EBIO activated apical Cl^– channels and basolateral K⁺ channels (presumably those that are Ca²⁺ dependent), thereby suggesting that this compound, or related derivatives, may be suitable as topical agents to stimulate fluid transport across the tissue in individuals with lacrimal gland deficiencies. Ussing chamber; short-circuit current; electrolyte transport; chloride secretagogue; potassium conductance; 1-ethyl-2-benzimidazolinone; 1,10-phenanthroline     

Potassium Channels at Chara Plasmalemma

Exposure to high K⁺ medium transforms Chara plasmalemma into[K⁺]_osensitive state (K⁺ state). The current-voltage (I/V)characteristicsunder such conditions display a negative conductance region.This feature results from the complex time and voltage dependenceof K⁺ channel opening At potentials more negative than a thresholdp.d. the channels are closed and the I/V characteristics becomelinear with a low slope conductance of 0.8 S m² and only a weakdependence on [K⁺]_o. Such behaviour is usually associated witha non-specific leak current The threshold level for K⁺ channelclosing depends on [K⁺]_o. In 2.0 mol m^–3 and 5.0 mol m^–3K⁺ medium the membrane resting p.d. follows E_K, but hyperpolarizesgradually if the [K⁺]_o is lowered. The proton pump thus appearsto be non-operative, while the cell is in the K⁺ state, andrecovers slowly as the cell is returned to a low K⁺ medium.Excitation currents decline if the cells are kept in K⁺ statefor some hours. Key words: K⁺ channels, Chara corallina, Proton pump, Current/, oltage characteristics, Conductance     

Protease-activated receptor regulation of Cl- secretion in Calu-3 cells requires prostaglandin release and CFTR activation

Human lung epithelial (Calu-3) cells were used to investigate the effects of protease-activated receptor (PAR) stimulation on Cl^– secretion. Quantitative RT-PCR (QRT-PCR) showed that Calu-3 cells express PAR-1, -2, and -3 receptor mRNAs, with PAR-2 mRNA in greatest abundance. Addition of either thrombin or the PAR-2 agonist peptide SLIGRL to the basolateral solution of monolayers mounted in Ussing chambers produced a rapid increase in short-circuit current (I_sc: thrombin, 21 ± 2 µA; SLIGRL, 83 ± 22 µA), which returned to baseline within 5 min after stimulation. Pretreatment of monolayers with the cell-permeant Ca²⁺-chelating agent BAPTA-AM (50 µM) abolished the increase in I_sc produced by SLIGRL. When monolayers were treated with the cyclooxygenase inhibitor indomethacin (10 µM), nearly complete inhibition of both the thrombin- and SLIGRL-stimulated I_sc was observed. In addition, basolateral treatment with the PGE₂ receptor antagonist AH-6809 (25 µM) significantly inhibited the effects of SLIGRL on I_sc. QRT-PCR revealed that Calu-3 cells express mRNAs for CFTR, the Ca²⁺-activated KCNN4 K⁺ channel, and the KCNQ1 K⁺ channel subunit, which, in association with KCNE3, is known to be regulated by cAMP. Stimulation with SLIGRL produced an increase in apical Cl^– conductance that was blocked in cells expressing short hairpin RNAs designed to target CFTR. These results support the conclusion that PAR stimulation of Cl^– secretion occurs by an indirect mechanism involving the synthesis and release of prostaglandins. In addition, PAR-stimulated Cl^– secretion requires activation of CFTR and at least two distinct K⁺ channels located in the basolateral membrane. cystic fibrosis transmembrane conductance regulator; KCNQ1; calcium-activated potassium channels; KCNN4; cAMP     

Efficiency of K+ Utilization by Barley Varieties: Activation of Pyruvate Kinase

Barley (Hordeum vulgare L.) varieties differed in their raponseto [K⁺]₀, in terms of their utilization efficiencies (UE = freshweight. concentration of [K⁺]₁^–1). At low [K⁺]₀, Compana,an efficient-non-responder demonstrated superior utilizationof absorbed K⁺. On the other hand, at high [K⁺]₀, Fergus (anefficient responder) and BT 334 (an inefficient responder) hadhigher UE values for K⁺ than Compana which performed poorlyat this [K⁺]₀. Kinetic parameters for K⁺ activation of the enzyme pyruvatekinase from 12 barley varieties, representing a range of UEvalues, were determined. Varieties showed substantial differencesin their V_max values (P<0·01). Compana, an efficientvariety, had the highest V_max (31 µmol g^–1 freshwt. h^–1) which was about 50% higher than that of Mingo,an inefficient variety. By contrast, K_m values for the enzymeswere not significantly different among varieties The mean valuesfor all varieties (3·9±0·15 mol m^–3K⁺) is far below the estimated cytoplasmic [K⁺] (100-200 molm^–3). It is, therefore, unlikely that differences in theutilization of K⁺ by these varieties can be explained on thebasis of differential requirements for (K⁺) activation of theseenzymes. Alternative possibilities for differences in the utilizationof K⁺ are discussed. Key words: K⁺ utilization efficiency, Pyruvate kinase, Barley varieties     

K+ Channel in the Chora Plasmalemma: Estimations of K+ Channel Density and Single K+ Channel Conductance

The electromotive force E and the conductance G of the Characorallina plasmalemma were measured under voltage clamp conditions.In the depolarized voltage range less negative than –60mV, E changed according to the Nerhst equation for K⁺, and Gincreased with the external K⁺ concentration [K⁺]_o and alsowith the depolarization of the membrane potential. This is attributedto the voltage-dependent opening of the K⁺ channels in the largelydepolarized voltage region. The voltage-dependent increase ofG was due to the increase of the number of open K⁺ channelsper unit area. The density of the total K⁺ channels in the C. corallina plasmalemmawas estimated to be about 6.50/(10 µm)2. The single K⁺channel conductance _K changed with the external [K⁺]_o; it was79.3, 86.1, 105.9, 119.0 pS for external [K⁺]_o of 0.2, 0.5,2.0 and 5.0 mu respectively. (Received May 22, 1986; Accepted August 22, 1986)     

Potassium-Ammonium Uptake Interactions in Tobacco Seedlings

Short-term (< 12 h) uptake experiments were conducted with6–7-week-old tobacco (Nicotiana tabacum L. cv. Ky 14)seedlings to determine absorption interactions between K⁺ andNH₄⁺. At equal solution concentrations (0.5 mol m^–3) netK⁺ uptake was inhibited 30–35% by NH₄⁺ and NH₄⁺ uptakewas decreased 9–24%. Removal of NH₄⁺ resulted in completerecovery in K⁺ uptake rate, but NH⁴⁺ uptake rate did not recoverwhen K⁺ was removed. In both cases, inhibition of the uptakerate of one cation saturated as the concentration of the othercation was increased up to 0.5 mol m^–3. The relative effectof K⁺-NH₄⁺ interactions was not altered when Cl- was replacedwith SO₄^2–, but the magnitudes of the uptake rates wereless in the absence of Cl-. The V_max for NH₄⁺ uptake was reducedfrom 128 to 105 µmol g^–1 dry wt. h^–1 in thepresence of 0.5 mol m^–3 K⁺ and the K_m for NH₄⁺ doubledfrom 12 to 27 mmol m^–3 in the presence of K⁺. The resultsof these K⁺-NH₄⁺ experiments are interpreted as mixed-noncompetitiveinteractions. However, an enhanced efflux of K⁺ coupled to NH₄⁺influx via an antiporter cannot be ruled out as contributingto the decrease in net K⁺ uptake. Key words: Nicotiana tabacum, K⁺, NH₄⁺, Uptake interactions     

Potassium Transport Across the Membranes of Chara: I. THE RELATIONSHIP BETWEEN RADIOACTIVE TRACER INFLUX AND ELECTRICAL CONDUCTANCE

Smith, J. R., Smith, F. A. and Walker, N. A. 1987. Potassiumtransport across the membrane of Chara. I. The relationshipbetween radioactive tracer influx and electrical conductance.—J.exp. Bot. 38:731–751. The ⁴²K influx () and the electrical conductance (G_m) were measured simultaneously for the ‘membrane’of internodal cells of Chara australis as a function of theexternal [KCl] (K?. In bathing solutions of pH = 5?0, progressively increased from 20?5to 430?60 nmol m^–2 s^–1 and G_m increased from 0?36?0?02to 3?8?0?8 S m^–2 when K? was increased from 0?1 to 10mol m^–3. The resting membrane potential difference (p.d.)was approximately -135 mV for low K? and approached the expectedNernst equilibrium p.d. for K⁺ ions when K? > 1?0 mol m^–3.Measurements of ³⁶Cl influx suggested that the ⁴²K influx waspredominantly electrogenic. The equivalent Goldman permeabilityto K⁺ ions (P_k) was approximately 20–30 nm s^–1 anddid not vary significantly with increasing K?. The equivalentconductance attributable to the electrogenic transport of K⁺ ions was calculated from assuming passive, independent diffusionof K⁺ ions and the ratio was found to be typically close to one. It was also found that themagnitudes of and G_m measuredsimultaneously for each individual cell were also well correlatedfor K? 1?0 mol m^–3, and that the slope of the line ofbest fit was close to one. For each K? it was found that theconductance not attributable to K⁺ translocation and presumablyassociated primarily with the transport of protons or theirequivalents was typically 0?2–0?5 Sm^–2. For K? >1?0 mol m^–3 the results indicated that the transport ofK⁺ ions was essentially independent, i.e. there was no evidencefor flux interactions. The results also indicated that the equivalentconductance derived from the measured ⁴²K influx could usefullyindicate the fraction of the electrical conductance attributableto the translocation of K⁺ ions. Key words: Potassium, conductance, influx     

Effect of irradiance and vapour pressure deficit On stomatal response to CO2 enrichment of four tree species

The stomatal response of seedlings grown in 360 or 720 µmolmol^–1 to irradiance and leaf-to-air vapour pressure deficit(VPD) at both 360 and 720 µmol mol^–1 to CO₂ wasmeasured to determine how environmental factors interact withCO₂ enrichment to affect stomatal conductance. Seedlings offour species with different conductances and life histories,Cercis canadensis (L.), Quercus rubra (L.), Populus deltoides(Bartr. ex Marsh.) P. nigra (L.), and Pinus taeda (L.), weremeasured in hopes of identifying general responses. Conductanceof seedlings grown at 360 and 720 µmol mol^–1 CO₂were similar and responded in the same manner to measurementCO₂ concentration, irradiance and VPD. Conductance was lowerfor all species when measured at 720 than when measured at 360µmol mol^–1 CO₂ at both VPDs ({small tilde}1.5 and{small tilde}2.5 kPa) and all measured irradiances greater thanzero (100, 300, 600,>1600 µmol m^–2 S^–2)The average decrease in conductance due to measurement in elevatedCO₂ concentration was 32% for Cercis, 29% for Quercus, 26% forPopulus, and 11% for Pinus. For alt species, the absolute decreasein conductance due to measurement in CO₂ enrichment decreasedas irradiance decreased or VPD increased. The proportional decreasedue to measurement in CO₂ enrichment decreased in three of eightcases: from 0.46 to 0.10 in Populus and from 0.18 to 0.07 inPinus as irradiance decreased from>1600 to 100 µmolm^–2 s^–1 and from 0.35 to 0.24 in Cercis as VPD increasedfrom 1.3 to 2.6 kPa. Key words: Stomatal conductance, CO₂ enrichment, irradiance, vapour pressure deficit     

Involvement of anion channel(s) in the modulation of the transient outward K(+) channel in rat ventricular myocytes

The cardiac Ca²⁺-independent transient outward K⁺ current (I_to), a major repolarizing ionic current, is markedly affected by Cl^– substitution and anion channel blockers. We reexplored the mechanism of the action of anions on I_to by using whole cell patch-clamp in single isolated rat cardiac ventricular myocytes. The transient outward current was sensitive to blockade by 4-aminopyridine (4-AP) and was abolished by Cs⁺ substitution for intracellular K⁺. Replacement of most of the extracellular Cl^– with less permeant anions, aspartate (Asp^–) and glutamate (Glu^–), markedly suppressed the current. Removal of external Na⁺ or stabilization of F-actin with phalloidin did not significantly affect the inhibitory action of less permeant anions on I_to. In contrast, the permeant Cl^– substitute Br^– did not markedly affect the current, whereas F^– substitution for Cl^– induced a slight inhibition. The I_to elicited during Br^– substitution for Cl^– was also sensitive to blockade by 4-AP. The ability of Cl^– substitutes to induce rightward shifts of the steady-state inactivation curve of I_to was in the following sequence: NO₃^– > Cl^– Br^– > gluconate^– > Glu^– > Asp^–. Depolymerization of actin filaments with cytochalasin D (CytD) induced an effect on the steady-state inactivation of I_to similar to that of less permeant anions. Fluorescent phalloidin staining experiments revealed that CytD-pretreatment significantly decreased the intensity of FITC-phalloidin staining of F-actin, whereas Asp^– substitution for Cl^– was without significant effect on the intensity. These results suggest that the I_to channel is modulated by anion channel(s), in which the actin cytoskeleton may be implicated. transient outward potassium current; anion channel; actin cytoskeleton; myocyte; potassium ion     

Effect of Root Zone Temperature on the Mineral Composition of Xylem Sap and Plasma Membrane K+-Mg++-ATPase Activity of Grafted-Cucumber and -Figleaf Gourd Root Systems

Cucumber (Cucumis sativus L.) seedlings were grafted onto cucumber-(CG) or figleaf gourd- (FG, Cucurbita ficifolia Bouché)seedlings in order to determine the effect of solution temperature(12, 22, and 32°C) on the mineral composition of xylem sapand the plasma membrane K⁺-Mg⁺⁺-ATPase activities of the roots.Low solution temperature (12°C) lowered the concentrationof NO^–₃ and H₂PO^–₄ in xylem sap of CG plants butnot of FG plants. Concentrations of K⁺, Ca⁺⁺ and Mg⁺⁺ in xylemsap were less affected than anions by solution temperature.The plasma membrane of FG plants grown in 12°C solutiontemperature showed the highest K⁺- Mg⁺⁺-ATPase activity at allATP concentrations up to 3 mM and at low reaction temperatureup to 12°C, indicating resistance of figleaf gourd to lowroot temperature. (Received December 27, 1994; Accepted March 10, 1995)     

Distinct K+ conductive pathways are required for Cl- and K+ secretion across distal colonic epithelium

Secretion of Cl^– and K⁺ in the colonic epithelium operates through a cellular mechanism requiring K⁺ channels in the basolateral and apical membranes. Transepithelial current [short-circuit current (I_sc)] and conductance (G_t) were measured for isolated distal colonic mucosa during secretory activation by epinephrine (Epi) or PGE₂ and synergistically by PGE₂ and carbachol (PGE₂ + CCh). TRAM-34 at 0.5 µM, an inhibitor of K_Ca3.1 (IK, Kcnn4) K⁺ channels (H. Wulff, M. J. Miller, W. Hänsel, S. Grissmer, M. D. Cahalan, and K. G. Chandy. Proc Natl Acad Sci USA 97: 8151–8156, 2000), did not alter secretory I_sc or G_t in guinea pig or rat colon. The presence of K_Ca3.1 in the mucosa was confirmed by immunoblot and immunofluorescence detection. At 100 µM, TRAM-34 inhibited I_sc and G_t activated by Epi (4%), PGE₂ (30%) and PGE₂ + CCh (60%). The IC₅₀ of 4.0 µM implicated involvement of K⁺ channels other than K_Ca3.1. The secretory responses augmented by the K⁺ channel opener 1-EBIO were inhibited only at a high concentration of TRAM-34, suggesting further that K_Ca3.1 was not involved. Sensitivity of the synergistic response (PGE₂ + CCh) to a high concentration TRAM-34 supported a requirement for multiple K⁺ conductive pathways in secretion. Clofilium (100 µM), a quaternary ammonium, inhibited Cl^– secretory I_sc and G_t activated by PGE₂ (20%) but not K⁺ secretion activated by Epi. Thus Cl^– secretion activated by physiological secretagogues occurred without apparent activity of K_Ca3.1 channels but was dependent on other types of K⁺ channels sensitive to high concentrations of TRAM-34 and/or clofilium. epinephrine; prostaglandin E₂; cholinergic; Kcnn4; TRAM-34; clofilium     

Further Characteristics of Salt-Dependent Bicarbonate Use by the Seagrass Zostera muelleri

Millhouse, J. and Strother, S. 1987. Further characteristicsof salt-dependent bicarbonate use by the seagrass Zostera muelleri.—J.exp. Bot. 38: 1055–1068. The contribution of HCO^–₃to photosynthetic O₂ evolutionin the seagrass Zostera muelleri Irmisch ex Aschers. increasedwith increasing salinity of the bathing seawater when the inorganiccarbon concentration was kept constant. K_1/2 (seawater salts)for HCO^–₃ -dependent photosynthesis was 66% of seawatersalinity. Both short- and long-term pretreatment at low salinitiesstimulated photosynthesis in full strength seawater. Twentyfour hours pre-incubation of seagrass plants in 3·0 molm^–3 NaHCO₃ resulted in increased photosynthesis at allsalinities, apparently due to stimulation of HCO^–₃ use(K_1/2 (seawater salts) = 26%). V_max (HCO^–₃) was not affectedby low salinity pretreatment. The kinetics of HCO^–₃ stimulationby the major seawater cations was investigated. Ca²⁺ was themost effective cation with the highest V_max (HCO^–₃) andwith K_1/2(Ca²⁺) = 14 mol m^–3. Mg²⁺ was also very effectiveat less than 50 mol m^–3 but higher concentrations wereinhibitory. This inhibition cannot be accounted for solely byprecipitation of MgCO₃. Na⁺ and K⁺ were both capable of stimulatingHCO^–₃ use. Stimulation was in two distinct parts. Up to500 mol m^–3, both citrate and chloride salts gave similarresults (K_1/2(Na⁺) 81 mol m^–3, V_max(HCO^–₃) 0·26µmol O₂ mg^–1 chl min^–1), but use of citratesalts above 500 mol m^–2 caused a second stimulation ofHCO^–₃ use (K_1/2(Na⁺) 830 mol m^–3, V_max(HCO^–₃)0·68 µmol O₂ mg^–1 chl min^–1). V_max(HCO^–₃)for the second-phase Na⁺ or K⁺ stimulation was of the same orderas for Ca²⁺-stimulated HCO^–₃ use. To further characterizesalt-dependent HCO^–₃ use, the sensitivity of photosynthesisto Tris and TES buffers was investigated. The effects of Trisappear to be due to the action of Tris⁺ causing stimulationof HCO^–₃ -dependent photosynthesis in the absence of salt,but inhibition of HCO^–₃ use in saline media. TES has noeffect on photosynthesis. External carbonic anhydrase, althoughimplicated in salt-dependent HCO^–₃ use in Z. muelleri,could not be detected in whole leaves. Key words: Zostera muelleri, HCO^–₃ use, salinity     

Purification, Properties and Phosphorylation of Anaerobically Induced Enolase in Echinochloa phyllopogon and E. crus-pavonis

Enolase (2-phospho-D-glycerate hydrolyase, EC 4.2.1.11 [EC] ) activityis differentially induced by anoxia in the flood-tolerant speciesE. phyllopogon (Stev.) Koss and the flood-intolerant speciesE. crus-pavonis (H.B.K.) Schult. To examine the regulation ofenolase at the protein level, we purified the enzyme from bothspecies to near homogeneity and compared their physico-chemicaland catalytic properties. Enolase purified from E. phyllopogonexhibits optimal activity at pH 7.0, a K_m of 80 µM for2-PGA, a Q₁₀ of 1.97 and an E_a of 12.3 kcal mol^-1. Similarly,enolase from E. crus-pavonis exhibits optimal activity at pH7.0, a K_m of 50 µM for 2-PGA, a Q₁₀ of 2.04 and an E_aof 12.9 kcal mol^-1. The enzyme from both species is thermostable(100% active after 15 min, 50°C) and is a homodimer of 52.5kDa subunits as resolved by SDS-PAGE and immunoblotting. E.phyllopogon enolase was phosphorylated in vitro using either[     

Dexamethasone treatment causes resistance to insulin-stimulated cellular potassium uptake in the rat

Patients treated with glucocorticoids have elevated skeletal muscle ouabain binding sites. The major Na⁺-K⁺-ATPase (NKA) isoform proteins found in muscle, ₂ and ₁, are increased by 50% in rats treated for 14 days with the synthetic glucocorticoid dexamethasone (DEX). This study addressed whether the DEX-induced increase in the muscle NKA pool leads to increased insulin-stimulated cellular K⁺ uptake that could precipitate hypokalemia. Rats were treated with DEX or vehicle via osmotic minipumps at one of two doses: 0.02 mg·kg^–1·day^–1 for 14 days (low DEX; n = 5 pairs) or 0.1 mg·kg^–1·day^–1 for 7 days (high DEX; n = 6 pairs). Insulin was infused at a rate of 5 mU·kg^–1·min^–1 over 2.5 h in conscious rats. Insulin-stimulated cellular K⁺ and glucose uptake rates were assessed in vivo by measuring the exogenous K⁺ infusion () and glucose infusion (G_inf) rates needed to maintain constant plasma K⁺ and glucose concentrations during insulin infusion. DEX at both doses decreased insulin-stimulated glucose uptake as previously reported. G_inf (in mmol·kg^–1·h^–1) was 10.2 ± 0.6 in vehicle-treated rats, 5.8 ± 0.8 in low-DEX-treated rats, and 5.2 ± 0.6 in high-DEX-treated rats. High DEX treatment also reduced insulin-stimulated K⁺ uptake. (in mmol·kg^–1·h^–1) was 0.53 ± 0.08 in vehicle-treated rats, 0.49 ± 0.14 in low-DEX-treated rats, and 0.27 ± 0.08 in high-DEX-treated rats. DEX treatment did not alter urinary K⁺ excretion. NKA ₂-isoform levels in the low-DEX-treated group, measured by immunoblotting, were unchanged, but they increased by 38 ± 15% (soleus) and by 67 ± 3% (gastrocnemius) in the high-DEX treatment group. The NKA ₁-isoform level was unchanged. These results provide novel evidence for the insulin resistance of K⁺ clearance during chronic DEX treatment. Insulin-stimulated cellular K⁺ uptake was significantly depressed despite increased muscle sodium pump pool size. skeletal muscle; sodium pump; Na⁺-K⁺-ATPase     

Efficiency of Potassium Utilization by Barley Varieties: The Role of Subcellular Compartmentation

Memon, A. R., Saccomani, M. and Glass, A. D. M. 1985. Efficiencyof potassium utilization by barley varieties: The role of subcellularcompartmentation.?J. exp. Bot. 36: 1860–1876. The subcellulardistributions of K⁺ in roots of three barley (Hordeum vulgareL.) varieties, grown at 10 and 100 mmol m^–3 external K⁺([K⁺]_o) were estimated by compartmental analyses. In general,increased [K⁺]_o caused a 2–3 fold increase in vacuolar[K⁺], but cytoplasmic [K⁺] increased only slightly. Nevertheless,the three varieties, which had been selected for study on thebasis of their different rates of K⁺ utilization, showed distinctdifferences in the allocation of K⁺ between cytoplasm and vacuole.At 10 mmol m^–3 [K⁺]_o var. Betzes exhibited typical K⁺deficiency symptoms while var. Fergus and var. Compana did not,even though Betzes had higher [K⁺] in shoots and roots. Theinefficient utilization of K⁺ in this variety appears to beassociated with a failure to mobilize vacuolar K⁺ into the cytoplasmiccompartment (the ratio of vacuolar: cytoplasmic K⁺ contentsfor Betzes was 4.1 compared to 2.7 and 2.5, respectively, forFergus and Compana). Fergus and Betzes, which demonstrate pronouncedgrowth responses to increased [K⁺]₀ between 10 and 100 mmolm^–3, showed significant increases of cytoplasmic [K⁺]in this range of [K⁺]_o. By contrast, cytoplasmic [K⁺] in Compana,a variety whose growth is not stimulated by increased [K⁺]₀(from 10 to 100 mmol m^–3) showed virtually no increase.It is suggested that the efficiency of K⁺ utilization and thegrowth response to [K⁺]₀ in these varieties are functions ofthe subcellular distribution of this ion between cytoplasm andvacuole. Key words: Barley varieties, K⁺ subcellular compartmentation, utilization efficiency     

Response of Photosynthesis, Stomatal Conductance and Water Use Efficiency to Elevated CO2 and Nutrient Supply in Acclimated Seedlings of Phaseolus vulgaris L.

Plants of Phaseolus vulgaris L were grown from seed in open-topgrowth chambers at present day (350 µmol mol^–1)and double the present day (700 µmol mol^–1) atmosphericCO₂ concentration with either low (L, without additional nutrientsolution) or relatively high (H, with additional nutrient solution)nutrient supply Measurements of assimilation rate, stomatalconductance and water use efficiency were started 17 d aftersowing on each fully expanded, primary leaf of three plantsper treatment Measurements were made in external CO₂ concentrations(C₂) of 200, 350, 450, 550 and 700 µmol mol^–1 andrelated to both C_a and to C₁, the mean intercellular space CO₂concentration Fully adjusted, steady state measurements weremade after approx 2 h equilibration at each CO₂ concentration The rate of CO₂ assimilation by leaves increased and stomatalconductance decreased similarly over the range of C_a or C₁ inall four CO₂ and nutrient supply treatments but both assimilationrate and stomatal conductance were higher in the high nutrientsupply treatment than in the low nutrient treatment The relationbetween assimilation rate or stomatal conductance and C₁ wasnot significantly different amongst plants grown in present-dayor elevated CO₂ concentration in either nutrient supply treatment,i e there was no evidence of down regulation of photosynthesisor stomatal response Increase in CO₂ concentration from 350to 700 µmol mol^–1 doubled water use efficiency ofindividual leaves in the high nutrient supply treatment andtripled water use efficiency in the low nutrient supply treatment The results support the hypothesis that acclimation phenomenaresult from unbalanced growth that occurs after the seed reservesare exhausted, when the supply of resources becomes growth limiting CO₂ enrichment, Phaseolus vulgaris L., net CO₂ assimilation rate, stomatal conductance, water use efficiency     

Ionic Currents across the Plasmalemma of Chara inflata Cells: III. WATER-RELATIONS PARAMETERS AND THEIR CORRELATION WITH MEMBRANE ELECTRICAL PROPERTIES

The water-relations parameters of Chara inflata cells were determineddirectly using the micro pressure probe technique. The turgorpressure of cells in artificial pond water (₀ = 0.06 MPa) wasabout 0.65 MPa and the half-time (T_1/2) for water exchange wasabout 6.5 s. The calculated values of the hydraulic conductivity(L_P) were in the range 1–2 ? 10^–6m s^–1 (MPa)^–1.The volumetric elastic modulus () was 32.8 MPa for turgor rangingfrom 0.77 to 0.82 MPa. Large changes in the water-relations parameters and the electricalproperties of the membrane occurred when the turgor was decreasedto low values. These changes included: (i) a decrease in theT_1/2 for water exchange, (ii) an increase in L_P and (iii) depolarizationof the membrane potential difference (V_m). The micro pressure probe, which enabled the turgor pressureof the cell to be altered, was used in combination with thevoltage-clamp technique to determine the relationship betweenK⁺ and Cl^– conductances of the plasmalemma and the cellturgor. The K⁺ conductance increased reversibly as the turgorwas reduced in the range 0 to 0.6 MPa and the Cl^– -conductanceincreased as the turgor was reduced in the range 0.1 to 0.5MPa. It is suggested that these pressure-dependent K⁺ and Cl^–conductances may have a dual role in electrical events and thenon-electrical responses such as changes in the cell volume. Key words: Chara inflata, membrane conductances, ion channels, water-relations parameters     

External Potassium Supply is not Required for Root Growth in Saline Conditions: Experiments with Ricinus communis L. Grown in a Reciprocal Split-Root System

Ricinus communis L. (castor bean) plants were grown in the absence(control) and in the presence of 100molm^–3NaCl with areciprocal split-root system, in which K⁺ was supplied to oneand NO₃^– to the other part of the root system. In theseplants shoot and, to a lesser extent, total root growth wereinhibited compared to plants with non-split roots. Without andwith NaCl, growth of roots receiving NO₃^– but noK⁺ (‘minusK/plus N-roots’) was substantially more vigorous thanunder the reverse conditions (‘plus K/minus N-roots¹).100mol m^–3 NaCl inhibited growth of minus K/plus N-roots¹to the same extent as that of non-split roots, indicating thatexternally supplied K⁺ was not required for root growth undersaline conditions. In growth media without added K⁺ the rootdepleted the external low K ⁺ levels resulting from chemicalsdown to a minimum value C_mln (1.0 to 1.4 mmol m^–3); inthe presence of 100 mol m^–3 NaCl, C_min, was higher (10–18mmol m^–3) and resulted from an initial net loss of K ⁺.C_min, was pH-dependent The distribution of K⁺, Na⁺ and Mg²⁺along the root was measured. In meristematic root tissues, K⁺ concentrations were scarcely affected by external K⁺ or byNaCl, where Na ⁺ concentrations were low, but somewhat elevatedat low external K+ and/or high NaCl. In differentiated, vacuolatedtissues K ⁺ concentrations were low and Na⁺ concentrations high,if K ⁺ was not supplied externally and/or NaCl was present.The longitudinal distribution of ions within the root was usedto estimate cytoplasmic and vacuolar ion concentrations. Thesedata showed a narrow homoeostasis of cytoplasmic K+ concentrations(100–140 mol m^–3) independent of external K ⁺ supplyeven in the presence of 100 mol m ^–3 NaCl. CytoplasmicNa ⁺ concentrations were maintained at remarkably low levels.Hence, external K⁺ concentrations above C_min, were not requiredfor maintaining K/Na selectivity, i.e. for controlling Na⁺ entry.The results are discussed with regard to mechanisms of K/Naselectivity and to the importance of phloem import of K⁺ forsalt tolerance of roots and for cytoplasmic K⁺ homoeostasis. Key words: Ricinus communis, nitrate, potassium, root (split-root), salt tolerance, phloem transport     

Long Term Effects of High CO2 Concentration on Photosynthesis of Water Hyacinth (Eichhornia crassipes (Mart.) Solms)

The photosynthetic response to CO₂ concentration, light intensityand temperature was investigated in water hyacinth plants (Eichhorniacrassipes (Mart.) Solms) grown in summer at ambient CO₂ or at10000 µmol(CO₂) mol^–1 and in winter at 6000 µmol(CO₂)mol^–1 Plants grown and measured at ambient CO₂ had highphotosynthetic rate (35 µmo1(CO₂) m^–2 s^–1),high saturating photon flux density (1500–2000) µmolm^–2 s^–1 and low sensitivity to temperature in therange 20–40 °C. Maximum photosynthetic rate (63 µmol(CO₂)m^–2 s^–1) was reached at an internal CO₂ concentrationof 800 µmol mol^–1. Plants grown at high CO₂ in summerhad photosynthetic capacities at ambient CO₂ which were 15%less than for plants grown at ambient CO₂, but maximum photosyntheticrates were similar. Photosynthesis by plants grown at high CO₂and high light intensity had typical response curves to internalCO₂ concentration with saturation at high CO₂, but for plantsgrown under high CO₂ and low light and plants grown under lowCO₂ and high light intensity photosynthetic rates decreasedsharply at internal CO₂ concentrations above 1000 µmol^–1. Key words: Photosynthesis, CO₂, enrichment, Eichhornia crassipes