Cl Flux Research Articles

Lung Fluid Balance During Development

After completing this article, readers should be able to: 1. Describe the major driving forces for liquid formation in and liquid removal from the lumen of the fetal lung. 2. Delineate the role of a normal volume of liquid within the lumen of the fetal lung for normal lung growth during development. 3. Describe the effects of labor on Na,K-ATPase activity in lung epithelial cells and clearance of lung liquid after birth. 4. Explain the route by which most residual liquid drains from the lung. 5. List the conditions associated with preterm birth that may contribute to delayed removal of fetal lung liquid. Curious students of mammalian development recognized more than a century ago that the lungs are filled with liquid during fetal life, (1) but the origin of that liquid remained obscure until 1948, when an unexpected discovery by two French scientists dispelled the prevailing view that fetal lung liquid derived from intrauterine inhalation of amniotic fluid. In studies designed to explore the ontogeny of the pituitary-adrenal axis, tracheal ligation of fetal rabbits for 9 days resulted in fluid distention of the lungs. (2) This serendipitous finding, subsequently confirmed and embellished by others, (3)(4)(5) established that the fetal lung itself, rather than the amniotic sac, is the source of the liquid that fills the lung during development. This liquid forms a slowly expanding structural template that prevents collapse and promotes growth of the fetal lung. Subsequent studies, conducted mostly with sheep, showed that liquid in the lumen of the fetal lung forms as a result of chloride secretion in the respiratory epithelium, (6)(7) a process that can be inhibited by diuretics that block Na,K-2Cl cotransport. (8)(9) In vitro experiments using cultured explants of lung tissue and monolayers of epithelial cells harvested from human fetal lung have …

Effect of divalent cations on ion fluxes and leaf photochemistry in salinized barley leaves

Photosynthetic characteristics, leaf ionic content, and net fluxes of Na(+), K(+), and Cl(-) were studied in barley (Hordeum vulgare L) plants grown hydroponically at various Na/Ca ratios. Five weeks of moderate (50 mM) or high (100 mM) NaCl stress caused a significant decline in chlorophyll content, chlorophyll fluorescence characteristics, and stomatal conductance (g(s)) in plant leaves grown at low calcium level. Supplemental Ca(2+) enabled normal photochemical efficiency of PSII (F(v)/F(m) around 0.83), restored chlorophyll content to 80-90% of control, but had a much smaller (50% of control) effect on g(s). In experiments on excised leaves, not only Ca(2+), but also other divalent cations (in particular, Ba(2+) and Mg(2+)), significantly ameliorated the otherwise toxic effect of NaCl on leaf photochemistry, thus attributing potential targets for such amelioration to leaf tissues. To study the underlying ionic mechanisms of this process, the MIFE technique was used to measure the kinetics of net Na(+), K(+), and Cl(-) fluxes from salinized barley leaf mesophyll in response to physiological concentrations of Ca(2+), Ba(2+), Mg(2+), and Zn(2+). Addition of 20 mM Na(+) as NaCl or Na(2)SO(4) to the bath caused significant uptake of Na(+) and efflux of K(+). These effects were reversed by adding 1 mM divalent cations to the bath solution, with the relative efficiency Ba(2+)>Zn(2+)=Ca(2+)>Mg(2+). Effect of divalent cations on Na(+) efflux was transient, while their application caused a prolonged shift towards K(+) uptake. This suggests that, in addition to their known ability to block non-selective cation channels (NSCC) responsible for Na(+) entry, divalent cations also control the activity or gating properties of K(+) transporters at the mesophyll cell plasma membrane, thereby assisting in maintaining the high K/Na ratio required for optimal leaf photosynthesis.

On Membrane Motor Activity and Chloride Flux in the Outer Hair Cell: Lessons Learned from the Environmental Toxin Tributyltin

The outer hair cell (OHC) underlies mammalian cochlea amplification, and its lateral membrane motor, prestin, which drives the cell's mechanical activity, is modulated by intracellular chloride ions. We have previously described a native nonselective conductance (GmetL) that influences OHC motor activity via Cl flux across the lateral membrane. Here we further investigate this conductance and use the environmental toxin tributyltin (TBT) to better understand Cl-prestin interactions. Capitalizing on measures of prestin-derived nonlinear capacitance to gauge Cl flux across the lateral membrane, we show that the Cl ionophore TBT, which affects neither the motor nor GmetL directly, is capable of augmenting the native flux of Cl in OHCs. These observations were confirmed using the chloride-sensitive dye MQAE. Furthermore, the compound's potent ability, at nanomolar concentrations, to equilibrate intra- and extracellular Cl concentrations is shown to surpass the effectiveness of GmetL in promoting Cl flux, and secure a quantitative analysis of Cl-prestin interactions in intact OHCs. Using malate as an anion replacement, we quantify chloride effects on the nonlinear charge density and operating voltage range of prestin. Our data additionally suggest that ototoxic effects of organotins can derive from their disruption of OHC Cl homeostasis, ultimately interfering with anionic modulation of the mammalian cochlear amplifier. Notably, this observation identifies a new environmental threat for marine mammals by TBT, which is known to accumulate in the food chain.

Functional Comparison of Mouse slc26a6 Anion Exchanger with Human SLC26A6 Polypeptide Variants: DIFFERENCES IN ANION SELECTIVITY, REGULATION, AND ELECTROGENICITY

The unusually low 78% amino acid identity between the orthologous human SLC26A6 and mouse slc26a6 polypeptides prompted systematic comparison of their anion transport functions in Xenopus oocytes. Multiple human SLC26A6 variant polypeptides were also functionally compared. Transport was studied as unidirectional fluxes of (36)Cl(-), [(14)C]oxalate, and [(35)S]sulfate; as net fluxes of HCO(3)(-) by fluorescence ratio measurement of intracellular pH; as current by two-electrode voltage clamp; and as net Cl(-) flux by fluorescence intensity measurement of relative changes in extracellular and intracellular [Cl(-)]. Four human SLC26A6 polypeptide variants each exhibited rates of bidirectional [(14)C]oxalate flux, Cl(-)/HCO(3)(-) exchange, and Cl(-)/OH(-) exchange nearly equivalent to those of mouse slc26a6. Cl(-)/HCO(3)(-) exchange by both orthologs was cAMP-sensitive, further enhanced by coexpressed wild type cystic fibrosis transmembrane regulator but inhibited by cystic fibrosis transmembrane regulator DeltaF508. However, the very low rates of (36)Cl(-) and [(35)S]sulfate transport by all active human SLC26A6 isoforms contrasted with the high rates of the mouse ortholog. Human and mouse orthologs also differed in patterns of acute regulation. Studies of human-mouse chimeras revealed cosegregation of the high (36)Cl(-) transport phenotype with the transmembrane domain of mouse slc26a6. Mouse slc26a6 and human SLC26A6 each mediated electroneutral Cl(-)/HCO(3)(-) and Cl(-)/OH(-) exchange. In contrast, whereas Cl(-)/oxalate exchange by mouse slc26a6 was electrogenic, that mediated by human SLC26A6 appeared electroneutral. The increased currents observed in oocytes expressing either mouse or human ortholog were pharmacologically distinct from the accompanying monovalent anion exchange activities. The human SLC26A6 polypeptide variants SLC26A6c and SLC26A6d were inactive as transporters of oxalate, sulfate, and chloride. Thus, the orthologous mouse and human SLC26A6 proteins differ in anion selectivity, transport mechanism, and acute regulation, but both mediate electroneutral Cl(-)/HCO(3)(-) exchange.

Ongoing hydrothermal heat loss from the 1912 ash-flow sheet, Valley of Ten Thousand Smokes, Alaska

The June 1912 eruption of Novarupta filled nearby glacial valleys on the Alaska Peninsula with ash-flow tuff (ignimbrite), and post-eruption observations of thousands of steaming fumaroles led to the name ‘Valley of Ten Thousand Smokes’ (VTTS). By the late 1980s most fumarolic activity had ceased, but the discovery of thermal springs in mid-valley in 1987 suggested continued cooling of the ash-flow sheet. Data collected at the mid-valley springs between 1987 and 2001 show a statistically significant correlation between maximum observed chloride (Cl) concentration and temperature. These data also show a statistically significant decline in the maximum Cl concentration. The observed variation in stream chemistry across the sheet strongly implies that most solutes, including Cl, originate within the area of the VTTS occupied by the 1912 deposits. Numerous measurements of Cl flux in the Ukak River just below the ash-flow sheet suggest an ongoing heat loss of ∼250 MW. This represents one of the largest hydrothermal heat discharges in North America. Other hydrothermal discharges of comparable magnitude are related to heat obtained from silicic magma bodies at depth, and are quasi-steady on a multidecadal time scale. However, the VTTS hydrothermal flux is not obviously related to a magma body and is clearly declining. Available data provide reasonable boundary and initial conditions for simple transient modeling. Both an analytical, conduction-only model and a numerical model predict large rates of heat loss from the sheet 90 years after deposition.

Effect of mercuric chloride on electrical parameters and anion fluxes in the toad skin

The amphibian skin, widely used for studying the transepithelial passage of electrolytes, exhibits anion pathways relatively specific for Cl −. We studied the effect of HgCl 2, 1.0×10 −4 M on its electrical parameters and unidirectional anion fluxes. In the presence of Cl −, the transepithelial conductance (G) of the isolated skin of the Bufo arenarum toad increased considerably following exposure to HgCl 2, whereas short-circuit current (SCC)—reflecting transepithelial Na + transport—underwent only slight stimulation. Following the blockade of Na + intake by amiloride, 1.0×10 −4 M, the removal of Cl − from the solution bathing the epidermal border of the skin brought about a decrease in G, and gave rise to a gradient-induced SCC (SCCg) consistent with transepithelial passage of Cl − along its gradient. Addition of mercaptoethanol, 5.0×10 −3 M to the bath containing Hg 2+ fully reversed these effects. The increase in G was accompanied by an increase in the unidirectional (epidermal to dermal) fluxes of 36Cl − and 131I −, and a decrease in the passage of 99mTcO 4 −. These results show the effects of HgCl 2 to be similar to those of theophylline, although exhibiting a different selectivity. Our data suggest that anion passage following exposure to HgCl 2 is, like that stimulated by theophylline, predominantly if not exclusively transcellular, and does not involve a significant opening of the tight junctions.

Effect of aluminium on membrane potential and ion fluxes at the apices of wheat roots.

Aluminium (Al) tolerance in wheat (Triticum aestivum L.) is associated with the Al-activated efflux of malate and K+ from the root apices. We tested the hypothesis that these Al-activated ion fluxes would induce changes in the membrane potential (Vm) and that these responses would differ between wheat genotypes that differ in Al tolerance. Within minutes of exposing wheat roots to 50 μm AlCl3, a significant depolarisation was measured in the Al-tolerant ET8 genotype but not in a near-isogenic, Al-sensitive genotype, ES8. We investigated the ion movements that may be responsible for these changes in Vm by measuring real-time fluxes of Cl-, H+ and K+ at the root apices of wheat seedlings using the non-invasive microelectrode ion flux estimation (MIFE) technique. Addition of 50 μm AlCl3 to the bathing solution stimulated an increase in K+ efflux and H+ influx in ET8 but not in ES8. The differences between the genotypes were sustained for 24 h and were observed only at the elongating zone and not the meristematic zone. After 24 h Al increased Cl- influx in ET8 but inhibited ES8 influx in a dose-dependent manner. These results provide new temporal and spatial information on the Al-activated ion fluxes from intact wheat plants.

Vasopressin‐stimulated CFTR Cl− currents are increased in the renal collecting duct cells of a mouse model of Liddle's syndrome

Liddle's syndrome is a genetic form of hypertension linked to Na(+) retention caused by activating mutations in the COOH terminus of the beta or gamma subunit of the epithelial sodium channel (ENaC). In this study, we used the short-circuit current (I(sc)) method to investigate the effects of deamino-8-d-arginine vasopressin (dDAVP) on Na(+) and Cl(-) fluxes in primary cultures of cortical collecting ducts (CCDs) microdissected from the kidneys of mice with Liddle's syndrome carrying a stop codon mutation, corresponding to the beta-ENaC R(566) stop mutation (L) found in the original pedigree. Compared to wild-type (+/+) CCD cells, untreated L/+ and L/L CCD cells exhibited 2.7- and 4.2-fold increases, respectively, in amiloride-sensitive (Ams) I(sc), reflecting ENaC-dependent Na(+) absorption. Short-term incubation with dDAVP caused a rapid and significant increase (approximately 2-fold) in Ams I(sc) in +/+, but not in L/+ or L/L CCD cells. In sharp contrast, dDAVP induced a greater increase in 5-nitro-2-(3-phenylpropamino)benzoate (NPPB)-inhibited apical Cl(-) currents in amiloride-treated L/L and L/+ cells than in their +/+ counterparts. I(sc) recordings performed under apical ion substituted conditions revealed that the dDAVP-stimulated apical secretion of Cl(-), which was absent in cultured CCDs lacking CFTR, was 1.8-fold greater in L/+ and 3.7-fold greater in L/L CCD cells than in their +/+ CCD counterparts. After the basal membrane had been permeabilized with nystatin and a basal-to-apical Cl(-) gradient had been imposed, dDAVP also stimulated larger Cl(-) currents across L/L and L/+ CCD layers than +/+ CCD layers. These findings demonstrate that vasopressin stimulates greater apical CFTR Cl(-) conductance in the renal CCD cells of mice with Liddle's syndrome than in wild-type mice. This effect could contribute to the enhanced NaCl reabsorption observed in the distal nephron of patients with Liddle's syndrome.

The co-crystallization of beryl-structure compounds in the Al 2Be 3Si 6O 18—Mg,Ca/F,Cl system

An Al 2Be 3Si 6O 18 — Mg, Ca/F, Cl flux system unused earlier has been chosen for the synthesis of beryl (Al 2Be 3Si 6O 18) and its varieties. In such a case of disregarding the well-known rules of the choice of a solvent, the following crystallization features of beryl have been revealed: (1) the beryl co-crystallizes successively and/or simultaneously with isostructural beryllian indialite (Mg 2BeAl 2Si 6O 18); (2) the bulk of beryl crystals is formed only in regions with high concentration of a flux; (3) the solution-melt liquates; (4) the solvent evaporates intensively. The advantages of the flux proposed are both the possibility of using inexpensive steel crucibles and good chromophoric properties of Cr, V, and Ti. Ratios of constitutient elements and some additive those between beryl and beryllian indialite, both grown simultaneously, and between pinacoid and prism in crystals of these compounds are reported.

Combined discriminative stimulus effects of midazolam with other positive GABAA modulators and GABAA receptor agonists in rhesus monkeys

Interactions among compounds at GABA(A) receptors might have important implications for the therapeutic and other effects of positive GABA(A) modulators (e.g. benzodiazepines). This study examined whether a midazolam discriminative stimulus is modified by GABA(A) agonists that act at sites other than benzodiazepine sites. Rhesus monkeys discriminating midazolam (0.32 mg/kg SC) received direct-acting GABA(A) receptor agonists (e.g. muscimol and gaboxadol), an indirect-acting GABA(A) receptor agonist (progabide), ethanol, another benzodiazepine (triazolam), a barbiturate (pentobarbital), or a neuroactive steroid (pregnanolone) alone and in combination with midazolam. When administered alone, triazolam (0.1 mg/kg), pentobarbital (17.8 mg/kg) and pregnanolone (5.6 mg/kg) occasioned high levels of midazolam lever responding, ethanol (1-3 g/kg) occasioned intermediate levels of midazolam lever responding, and muscimol (0.32-1 mg/kg), gaboxadol (3.2-10 mg/kg) and progabide (10-32 mg/kg) occasioned low levels of midazolam lever responding. When combined with less-than-fully effective doses of midazolam, progabide (32 mg/kg) and ethanol (1 g/kg), but not muscimol and gaboxadol, enhanced the midazolam discriminative stimulus. Triazolam, pregnanolone and pentobarbital increased the potency of midazolam to occasion midazolam lever responding and the effects of these combinations were additive. Direct-acting GABA(A) receptor agonists are qualitatively different from positive GABA(A) modulators in rhesus monkeys trained to discriminate midazolam. Although GABA(A) receptor agonists and modulators can enhance the actions of benzodiazepines at the GABA(A) receptor complex, the same drugs do not necessarily modify the discriminative stimulus effects of benzodiazepines. These results underscore the importance of the mechanism by which drugs alter Cl(-) flux at the GABA(A) receptor complex as a determinant not only of drug action but also of drug interaction and whether any particular drug enhances the behavioral effects of a benzodiazepine.

Water-soluble chlorides in massive seafloor serpentinites: a source of chloride in subduction zones

Dissolved cation and anion species were measured in the water-soluble fraction of massive seafloor serpentinites leached with distilled H2O. Soluble chloride contents ranged from 0.09 to 0.51 wt.% (average=0.26 wt.%), SO42− to 0.09 wt.%, Mg2+ to 0.16 wt.% and Na+ to 0.09 wt.%. The concentration of evolved ions was the same in samples equilibrated with water at 25 and 230 °C, demonstrating that a significant amount of Cl present in serpentinites occurs as water-soluble salts. A nearly equal amount of Cl exists in the leached serpentinite, likely sited in serpentinite itself. The formation of intergranular salts in the serpentinite can be explained by seawater hydrothermal alteration of peridotite. Hydration reactions and H2O-incorporation in the serpentine leave the residual fluid with ever-increasing salinity and ultimately lead to precipitation of water-soluble chlorides. Chlorine isotope ratios in the water-soluble fraction average −1.49‰ (vs. standard mean ocean chlorine, SMOC) from a core at the Iberian Abyssal Plain, and +0.16‰ from a much younger core at Hess Deep. The δ37Cl value of insoluble Cl is ∼0.33‰ higher than the soluble fraction. The nonzero δ37Cl values demonstrate that the Cl in the serpentinites is not an artifact of seawater incorporation during drilling.The following conclusions are presented: (1) Serpentinite may be the major conduit for surficial chlorine transfer to mantle depths and an important part of the global chlorine cycle. The Cl flux in serpentinites into the mantle is larger than all other previously identified fluxes. (2) Cl is sited in both a water-soluble and -insoluble component. (3) A high-salinity fluid ‘plume’ is evolved during serpentinite dehydration of a subducting slab. The high-salinity fluid affects metamorphic reactions and melting properties above the slab and can cause intense metasomatism. The unusual bulk composition of the whiteschist from Dora Maira can be explained by channeled passage of a high-salinity fluid. (4) High-salinity fluids reduce the stability field of serpentine, causing its dehydration to occur over a broad interval, which may partly explain the broad locus of intermediate-depth earthquakes.

Variation in throughfall deposition across a deciduous beech ( Fagus sylvatica L.) forest edge in Flanders

Throughfall deposition and canopy exchange of acidifying and eutrophying compounds and major base cations were studied by means of throughfall analysis in a deciduous beech ( Fagus sylvatica L.) forest edge in Belgium over a period of 1 year. Throughfall fluxes of Cl −, NH 4 + and Na + were significantly elevated at the forest edge compared to the forest interior. As no edge effect on throughfall water volume could be detected, the observed edge enhancement effects were mainly due to dry deposition and canopy exchange patterns. Indeed, there was an elevated dry deposition of Cl −, Na +, K +, Ca 2+ and Mg 2+ up to 50 m from the field/forest border. Within the forest, throughfall and dry deposition of SO 4 2− were highly variable and no significant differences were observed between the forest edge and the forest interior. Leaching of K + and Ca 2+ was reduced in the forest edge up to a distance of 30 m from the border. The measured nitrogen and acidic depositions far exceeded the current Flemish critical loads with respect to the protection of biodiversity in forests, especially at the forest edge. This points to an urgent need for controlling emissions as well as the need to consider the elevated deposition load in forest edges when calculating the critical loads in forests.

Identification of CFTR activators and inhibitors: chance or design?

The cystic fibrosis transmembrane conductance regulator (CFTR) is a cAMP-activated Cl(-) channel expressed in various epithelial cells, and is a pharmacological target for activators and inhibitors. Activators are useful for the pharmacotherapy of cystic fibrosis, specifically for those mutations that affect CFTR protein by reducing its ability to stay in the open state. Conversely, inhibitors are potentially useful to treat secretory diarrhoea caused by enterotoxins, as the CFTR is the main route for Cl(-) flux in the intestine. Recently, a variety of potent modulators of the CFTR Cl(-) channel activity have been identified by high-throughput screening of a large collections of small molecules. The identification of CFTR activators and inhibitors with novel chemical scaffolds might help with the rational design of compounds with improved pharmacological properties.

4-Chloro-benzo[F]isoquinoline (CBIQ) activates CFTR chloride channels and KCNN4 potassium channels in Calu-3 human airway epithelial cells.

1 Calu-3 cells have been used to investigate the actions of 4-chloro-benzo[F]isoquinoline (CBIQ) on short-circuit current (SCC) in monolayers, whole-cell recording from single cells and by patch clamping. 2 CBIQ caused a sustained, reversible and repeatable increase in SCC in Calu-3 monolayers with an EC50 of 4.0 microm. Simultaneous measurements of SCC and isotopic fluxes of 36Cl- showed that CBIQ caused electrogenic chloride secretion. 3 Apical membrane permeabilisation to allow recording of basolateral membrane conductance in the presence of a K+ gradient suggested that CBIQ activated the intermediate-conductance calcium-sensitive K(+)-channel (KCNN4). Permeabilisation of the basolateral membranes of epithelial monolayers in the presence of a Cl- gradient suggested that CBIQ activated the Cl(-)-channel CFTR in the apical membrane. 4 Whole-cell recording in the absence of ATP/GTP of Calu-3 cells showed that CBIQ generated an inwardly rectifying current sensitive to clotrimazole. In the presence of the nucleotides, a more complex I/V relation was found that was partially sensitive to glibenclamide. The data are consistent with the presence of both KCNN4 and CFTR in Calu-3. 5 Isolated inside-out patches from Calu-3 cells revealed clotrimazole-sensitive channels with a conductance of 12 pS at positive potentials after activation with CBIQ and demonstrating inwardly rectifying properties, consistent with the known properties of KCNN4. Cell-attached patches showed single channel events with a conductance of 7 pS and a linear I/V relation that were further activated by CBIQ by an increase in open state probability, consistent with known properties of CFTR. It is concluded that CBIQ activates CFTR and KCNN4 ion channels in Calu-3 cells.

Phenotypic differences in cholinergic responses of distal colonic epithelium.

The Flinders sensitive line (FSL) rats exhibit an increased cholinergic responsiveness in vivo when compared to their counterparts, the Flinders resistant line (FRL) rats. The functional consequences of this phenotypic difference on colonic mucosal function are not known. We sought to determine whether isolated distal colonic mucosa from the two strains exhibit differential responses to cholinergic agonists. The responses of the distal colonic mucosa from two lines of rats to carbachol were compared by recording changes in short-circuit current. The ion movements associated with these changes were assessed by flux analysis of the radiotracers, 22Na and 36Cl. The anticipated hyper-responsiveness to cholinergic stimulation in FSL rats was not seen. Carbachol responses were significantly enhanced by indomethacin pretreatment only in FRL rats. Tetrodotoxin (TTX) pretreatment significantly reduced responses to carbachol in FSL rats at all concentrations tested, though this was only seen with lower concentrations in FRL rats. Flux analysis indicated that both lines absorbed Na+ and Cl- under basal conditions and that a significant residual flux was present. Stimulation with carbachol led to significant reductions in net Na+ and Cl- fluxes in both lines. The changes in net Na+ and Cl- flux in both lines stem largely from a decrease in mucosal to serosal fluxes of both ions with an increase in serosal to mucosal flux of Cl-. The striking difference is the significant reduction in residual flux seen only in FRL rats. Indomethacin pretreatment abolished the changes in residual flux seen in FRL rats. Thus the responses to carbachol in these rats had at least three components: (a) a direct effect on the transporting colonocyte, (b) an indirect effect mediated by an arachidonic acid metabolite, and (c) another indirect effect involving a neurotransmitter. The relative contributions of each of these components were different in the two lines.

WNK4 regulates apical and basolateral Cl- flux in extrarenal epithelia.

Mutations in the serine-threonine kinase WNK4 [with no lysine (K) 4] cause pseudohypoaldosteronism type II, a Mendelian disease featuring hypertension with hyperkalemia. In the kidney, WNK4 regulates the balance between NaCl reabsorption and K(+) secretion via variable inhibition of the thiazide-sensistive NaCl cotransporter and the K(+) channel ROMK. We now demonstrate expression of WNK4 mRNA and protein outside the kidney. In extrarenal tissues, WNK4 is found almost exclusively in polarized epithelia, variably associating with tight junctions, lateral membranes, and cytoplasm. Epithelia expressing WNK4 include sweat ducts, colonic crypts, pancreatic ducts, bile ducts, and epididymis. WNK4 is also expressed in the specialized endothelium of the blood-brain barrier. These epithelia and endothelium all play important roles in Cl(-) transport. Because WNK4 is known to regulate renal Cl(-) handling, we tested WNK4's effect on the activity of mediators of epithelial Cl(-) flux whose extrarenal expression overlaps with WNK4. WNK4 proved to be a potent inhibitor of the activity of both the Na(+)-K(+)-2Cl(-) cotransporter (NKCC1) and the Cl(-)/base exchanger SLC26A6 (CFEX) (>95% inhibition of NKCC1-mediated (86)Rb influx, P < 0.001; >80% inhibition of CFEX-mediated [(14)C] formate uptake, P < 0.001), mediators of Cl(-) flux across basolateral and apical membranes, respectively. In contrast, WNK4 showed no inhibition of pendrin, a related Cl(-)/base exchanger. These findings indicate a general role for WNK4 in the regulation of electrolyte flux in diverse epithelia. Moreover, they reveal that WNK4 regulates the activities of a diverse group of structurally unrelated ion channels, cotransporters, and exchangers.

Factors Regulating the Distribution of Elements in the Sediments of a Seasonally Anoxic Lake in Tropical Taiwan

Sixteen metals, both total and acid-leached, and their binding formations were quantified for the sediment column of the seasonally anoxic Great Ghost Lake. Since marked variation in the content of organic matter plays a major role in regulating the distribution of metals in sediments, this study provided insight into the relationship between organic matter and metals in a naturally anoxic water environment. Metals, Al, Cs, Mg, Rb and V, had significant negative correlations with total carbon content (TC) and occurred less often in mobile fractions. These findings indicate that these metals came from lithogenic sources and did not change much during early diagenesis. As a result, signals of these elements, whose origins can be traced back to long-range transport dust, were better preserved. Acid-leached contents of Cs, Mg and Rb also had negative correlations with TC, while both total and acid-leached As and Cd had good positive correlations with TC. Large portions of both As and Cd, were observed in mobile fractions (especially As) and came from organic sources and/or were incorporated into organic matter during early diagenesis (especially Cd). Long-term variations in redox conditions of the lake were induced by climate changes and might have influenced distributions of Sr and redox-sensitive Mn and V in mobile fractions. In addition to natural processes, enrichment of Pb and Cd near the core top was from anthropogenic aeolian sources. Finally, recent aeolian fluxes of Al, As, Ba, Br, Ca, Cd, Ce, Cl, Cr, Cs, Cu, Er, Eu, Fe, Ga, Gd, K, La, Lu, Mg, Mn, Na, Nd, Ni, Pb, Rb, Si, Sm, Sr, Ti, Tm, V, Yb, Zn and Zr were estimated and large amounts of micronutrient Fe (~6O μg cm^(-2) yr^(-1)), of aeolian origin, was also noted.

Cellular and whole-plant chloride dynamics in barley: insights into chloride-nitrogen interactions and salinity responses.

The first analysis of chloride fluxes and compartmentation in a non-excised plant system is presented, examining ten ecologically pertinent conditions. The short-lived radiotracer couple (38)Cl/(39)Cl was used as a Cl(-) tracer in intact barley ( Hordeum vulgare L. cv. Klondike) seedlings, which were cultured and investigated under four external [Cl(-)], from abundant (0.1 mM) to potentially toxic (100 mM). Chloride-nitrogen interactions were investigated by varying N source (NO(3)(-) or NH(4)(+)) and strength (0.1 or 10 mM), in order to examine, at the subcellular compartmentation level, the antagonism, previously documented at the influx level, between Cl(-) and NO(3)(-), and the potential role of Cl(-) as a counterion for NH(4)(+) under conditions in which cytosolic [NH(4)(+)] is excessive. Cytosolic [Cl(-)] increased with external [Cl(-)] from 6 mM to 360 mM. Cl(-) influx, fluxes to vacuole and shoot, and, in particular, efflux to the external medium, also increased along this gradient. Efflux reached 90% of influx at the highest external [Cl(-)]. Half-times of cytosolic Cl(-) exchange decreased between high-affinity and low-affinity influx conditions. The relationship between cytosolic [Cl(-)] and shoot flux indicated the presence of a saturable low-affinity transport system ("SLATS") responsible for xylem loading of Cl(-). N source strongly influenced Cl(-) flux to the vacuole, and moderately influenced Cl(-) influx and shoot flux, whereas efflux and half-time were insensitive to N source. Cytosolic pool sizes were not strongly or consistently influenced by N source, indicating the low potential for Cl(-) to act as a counterion to hyperaccumulating NH(4)(+). We discuss our results in relation to salinity responses in cereals.

Plasma etching of high dielectric constant materials on silicon in halogen chemistries

Plasma etching of ZrO2 and HfO2 was studied in BCl3/Cl2 plasmas, as functions of the ion energy, chamber pressure, microwave power, and gas compositions. MClx is found to be the major etching product in Cl2 plasmas while MBxCly is the major etching product in BCl3 plasmas. The etching selectivity to Si is increased at lower ion energies and higher electron temperatures. Increasing microwave powers and reducing chamber pressures in BCl3/Cl2 plasmas increased the Cl and BCl2+ densities in the gas phase and consequently increased the metal oxide etch rate. A phenomenological model that takes into account the Cl density, BCl2+ density, and metal oxygen bond strength is proposed to describe the etch rate of ZrO2 and HfO2 in pure BCl3 plasmas as functions of the ion energy, microwave power, and chamber pressure. More accurate Cl flux measurement is needed to improve the model predictions.

Net uptake of chloride across the posterior gills of the Chinese crab ( Eriocheir sinensis)

Two methods are commonly used for the determination of transbranchial net fluxes of Na + and Cl −: direct analysis of changes in ion concentrations in the external medium using flame spectrophotometry or titration (net flux method), and measurement of unidirectional ion fluxes by means of radioactive tracers (tracer method). When we applied both methods in the same preparation, the isolated perfused posterior gill of freshwater-acclimated Eriocheir sinensis, to determine net fluxes of Cl −, the results differed substantially. In artificial fresh water (AFW) containing NaCl, the net flux method yielded a net uptake, but the tracer method showed a net efflux of Cl −. The net uptake of Cl − was abolished in Na +-free AFW indicating that Cl − uptake is coupled with the uptake of Na +. Applying the tracer method, net efflux of Cl − remained almost unchanged in Na +-free AFW. This suggests the opposite mechanism, i.e. uncoupled uptake of Na + and Cl −. The discrepancy in the results obviously depends on the method employed. Since the data obtained with the net flux method explain the osmoregulatory performance of crabs living in fresh water, we consider this method as appropriate for determining net transbranchial ion fluxes.