Ca Pool Research Articles

Solubility of major cations and Cu, Zn and Cd in soil extracts of some contaminated agricultural soils near a zinc smelter in Norway: modelling with a multisurface extension of WHAM

SummaryMechanistic modelling offers a means of simulating the speciation and solubility of trace metals in soils. The WHAM/Model VI model has previously been used to simulate pH buffering and Al solubility in acid soils, and metal partitioning in very organic soils, but has not previously been applied to agricultural soils. Here we have extended the WHAM/Model VI framework to include surface complexation to oxides, and cation exchange, and applied it to batch titrations of limed agricultural soils contaminated by emissions of Cd and Zn from a metal smelter. In contrast to previous studies on forest soils, model predictions were most sensitive to the size of the geochemically ‘active’ soil Ca pool. Following optimization of this pool the model reproduced trends in pH and major cations well. Blind predictions of soil metal concentrations (Cu, Zn and Cd), from estimates of the active soil metal concentrations obtained by extraction with 0.22 m HNO3, were mostly very reasonable. Where predicted metal solubility was different, the model could be fitted to the data by optimizing the size of the active metal pool. In some cases the optimized metal pool was unrealistically large, which indicates a possible deficiency in the way the model considers binding activity and competition. Organic matter was the dominant binding phase in these soils. These results support the contention that speciation modelling has great promise in providing a holistic description of ion chemistry in soils for both major and trace elements.

Impact of Harvest Intensity on Long-Term Base Cation Budgets in Swedish Forest Soils

The effects of harvesting on the long-term mass balances of calcium (Ca), magnesium (Mg) and potassium (K) were evaluated on a regional level in Sweden. A new high-resolution weathering database was used together with estimates of total deposition, losses through harvest and leaching. Estimates were made for pine and spruce separately and for two harvesting intensity scenarios: stem harvesting and whole-tree harvesting. The mass balance calculations showed net losses of Ca and Mg in almost the whole country for both scenarios. The losses were smaller for pine than for spruce. The K balances were mainly positive for pine but negative for spruce. Leaching was a main factor in the mass balances, especially for Ca and Mg. Whole-tree harvesting in spruce forests led to substantially higher net losses of K and Ca than stem harvesting, according to the calculations. In the whole-tree harvesting scenario in spruce forests the estimated yearly net losses of Ca, Mg and K corresponded to at least 5%, 8% and 3% of the pools of exchangeable base cations, respectively, at 25% of the analysed sites. If losses of this magnitude continue the depletion of the pools of Ca, Mg and K may lead to very low base saturation of the soils, possibly accompanied by negative effects on soil fertility, runoff water quality, tree vitality and tree growth within a forest rotation in parts of Sweden. Avoiding whole-tree harvesting can improve the situation substantially for K, but the losses of Ca and Mg will still be significant.

Store-Operated Ca2+ Entry in Muscle Physiology

Store-operated Ca 2+ entry (SOCE) represents a unique Ca 2+ entry mechanism, where Ca 2+ channels located on the plasma membrane sense the Ca 2+ filling status of intracellular Ca 2+ stores and gate the entry of Ca 2+ from the extracel- lular reservoir to replenish intracellular Ca 2+ storage. This pathway has received great interest not only because of its un- usual nature as a retrograde signal, but also due to its wide occurrence in both excitable and non-excitable cells and its po- tential role in various physiological and pathophysiological situations. In skeletal muscle, contractility is contingent upon the maintenance of intracellular Ca 2+ homeostasis, which requires the preservation of low levels of resting cytosolic Ca 2+ , readily available releasable pool of Ca 2+ from the sarcoplasmic reticulum, as well as functional Ca 2+ uptake and extrusion mechanisms. Recent studies have demonstrated that SOCE is present in skeletal muscle, and may play a significant role in muscle physiology. While the need for SOCE is increased during strenuous muscle exercise and fatigue, disruption of this process can lead to pathophysiological conditions. Repressed SOCE activity has been linked to aging related dysfunction. Elevated SOCE could lead to elevated intracellular Ca 2+ in dystrophic muscle cells and progression of muscular dystro- phy. The role of SOCE in the physiology and pathophysiology of skeletal muscle is a subject of increasing interest in the muscle biology field. Manipulation of SOCE by chemical, pharmacological and genetic approaches should have great po- tential in the treatment of muscle disorders that involve dysfunctional Ca 2+ homeostasis.

Estimation of tree biomass of Norway spruce forest in the Plešné Lake catchment, the Bohemian Forest

This paper evaluates the total biomass and pools of major nutrients and ecologically important metals of the tree layer in the catchment of Plesne jezero (PL) in the Bohemian Forest (Sumava, Czech Republic), and compares them to analogous data on understory vegetation and soils. The results are based on field measurements and semi-automatic image analyses of aerial orthophotographs. The tree layer was relatively sparse with open canopy in some parts of the catchment. Stand density varied between 44 and 328 individuals per hectare. The catchment weighted mean total biomass of trees was 134 t ha−1 dry weight, of which needles, branches, roots, and stems represented 5%, 10%, 14%, and 71%, respectively. The stem wood and bark represented 67% and 4%, respectively, of the total tree biomass. The catchment weighted mean element pools were 568 and 3.0 mol m−2 (i.e., 68 and 0.42 t ha−1) for C and N, respectively. The other pools were 76 mmol P m−2, 602 mmol Ca m−2, 133 mmol Mg m−2, 39 mmol Na m−2, 347 mmol K m−2, 19 mmol Al m−2, 6.2 mmol Fe m−2, and 35 mmol Mn m−2. The element pools accumulated in the tree biomass represented from < 1% (Al, Fe) to 37% (C) of their total pools (soil + tree layer + understory vegetation) in the catchment. Pools of Ca and Mg in the tree biomass were similar to their exchangeable pools in the catchment soils, while those of K were 3 times higher. Nutrient (N, P, Ca, Mg, and K) and C pools in the tree biomass were 2–11 times higher than those in the understory vegetation, with the minimum for P and maximum for C.

Manganese suppresses ATP-dependent intercellular calcium waves in astrocyte networks through alteration of mitochondrial and endoplasmic reticulum calcium dynamics

The neurotoxicity of manganese [Mn] is due in part to glutamate excitotoxicity. Release of ATP by astrocytes is a critical modulator of glutamatergic neurotransmission, which is regulated by calcium (Ca 2+) waves that propagate through astrocytic networks in response to synaptic activity. It was postulated that Mn alters ATP-dependent intracellular Ca 2+ dynamics in astrocytes, thereby suppressing Ca 2+ wave activity. Confluent primary cultures of cortical astrocytes were loaded with the Ca 2+-sensitive dye fluo-4 and examined by fluorescence microscopy for Ca 2+ wave activity following micropipet mechanical stimulation of a single cell. Mitochondrial Ca 2+ was evaluated by fluorescence microscopy following addition of ATP using the mitochondrial-specific Ca 2+ dye rhod-2-AM. Imaging studies revealed that pretreatment of astrocytes with 1–10 μM Mn significantly reduced the rate, area, and amplitude of mechanically induced Ca 2+ waves. This attenuation was not a result of inhibited mitochondrial calcium uptake because robust calcium waves were still observed following pretreatment of astrocytes with Ru360, an inhibitor of mitochondrial Ca 2+ uptake, either in coupling or uncoupling conditions. However, determination of endoplasmic reticulum (ER) Ca 2+ levels in cells using the sarco/endoplasmic reticulum Ca 2+-ATPase inhibitor thapsigargin indicated that Mn reduced the available pool of releasable ER Ca 2+ at concentrations as low as 1 μM. Examination of ATP-stimulated changes in mitochondrial Ca 2+ indicated that, in cells pretreated with Mn, mitochondria retained high levels of Ca 2+. It is concluded that exposure of astrocytes to low concentrations of Mn 2+ results in sequestration of Ca 2+ within the mitochondria that reduces the available pool of releasable Ca 2+ within the ER, thereby inhibiting calcium wave activity.

Thymosin β4 and AcSDKP inhibit the proliferation of HL‐60 cells and induce their differentiation and apoptosis

Our previous works have shown that bone marrow stromal cells secrete thymosin beta4 (Tbeta4) and AcSDKP. Tbeta4 and AcSDKP are existed in the conditioned medium of bone marrow endothelial cells. They exerted inhibitory effects on hematopoietic cells and then had protective effect on the early hematopoietic cells, which were cultured in the presence of hematopoietic stimulators. Thymosin beta4 consists of 43 peptides with a molecular weight of 4963. It contains at its N-terminal end the sequence of the acetylated tetrapeptide Ac-N-Ser-Asp-Lys-Pro (AcSDKP). This study was performed to evaluate the effect of Tbeta4 and AcSDKP on the growth of HL-60 cells. It was showed that Tbeta4 (10(-11)-10(-7)mol/L) and AcSDKP (10(-11)-10(-7)mol/L) had the dose-dependent inhibitory effect on the proliferation of HL-60 cells. Based on cell morphology and NBT reduction, Tbeta4 and AcSDKP induced differentiation of HL-60 cells. Morphologic and DNA fragment analysis proved that Tbeta4 and AcSDKP induced apoptosis of HL-60 cells. In order to analyze the mechanism of the effects of Tbeta4 and AcSDKP, intracellular free Ca(2+) concentration ([Ca(2+)](i)) of HL-60 leukemic cells was tested and Atlas cDNA Expression Array was performed. The results showed that Tbeta4 and AcSDKP could increased [Ca(2+)](i) by stimulating the release of Ca(2+) from intracellular Ca(2+) pool. Moreover, AcSDKP could also elicit a potent extracelluar calcium influx in HL-60 cells. Tbeta4 could also change apoptotic-related gene expression in leukemic cells, and resulted in the inhibition of proliferation and induction of differentiation and apoptosis of leukemic cells.

Two-week feeding of difructose anhydride III enhances calcium absorptive activity with epithelial cell proliferation in isolated rat cecal mucosa

Objective Ingestion of difructose anhydride III (DFAIII) enhances calcium (Ca) absorption in rats. The present study investigated the mechanism involved in increased Ca transport by DFAIII ingestion. The short-term and long-term effects of DFAIII feeding on Ca transport were determined by using isolated epithelium from the small and large intestine in rats. Methods Male Sprague-Dawley rats were fed an 8% cellulose or 5% cellulose plus 3% DFAIII diet for 14 d. Net epithelial Ca transport in the small intestine, cecum, and colon was compared between the two diet groups by using an Ussing chamber. The contents and epithelial tissues in the cecum were analyzed. Results There were no differences in basal and luminal DFAIII-induced Ca transport in the isolated small intestinal and colonic mucosa between the two diet groups. Basal and lumen DFAIII-induced Ca transport in the cecum in the DFAIII-fed group was higher than that in the control group. A decrease in pH and an increase in Ca pools, short-chain fatty acids, or organic acids in the cecal contents and in the depth and number of cells in crypts in cecal tissue were observed. Conclusions The increase in Ca transport involved two mechanisms: the presence of DFAIII in the small intestine directly affected the epithelial tissue and caused increased Ca absorption as a short-term effect, and the degradation of DFAIII by microbial fermentation produced short-chain fatty acids and subsequently enhanced Ca absorption in the large intestine as a long-term effect.

Effects of Thiol Chelation on α1-Adrenoceptor-Induced Vasoconstriction In Vivo

The aims of this study were to determine whether systemic injections of the lipophobic thiol chelator, para-hydroxymercurobenzoic acid (PHMBA) would reduce the vasoconstrictor responses elicited by the alpha1-adrenoceptor agonist, phenylephrine, in urethane-anesthetized rats by chelation of thiol residues in alpha1-adrenoceptors in vascular smooth muscle rather than voltage-sensitive Ca(2+)-channels (Ca(2+)VERSUS-channels). The magnitudes and durations of the vasoconstrictor responses elicited by phenylephrine were markedly reduced after the injections of PHMBA. In contrast, the maximal phenylephrine-induced responses were not affected whereas the durations of these responses were markedly attenuated after injection of the Ca(2+)VERSUS-channel blocker, nifedipine. Nifedipine elicited pronounced and sustained falls in mean arterial blood pressure and vascular resistances in PHMBA-treated rats. Moreover, the vasodilator actions of the nitric oxide-donor, sodium nitroprusside were minimally attenuated by PHMBA whereas they were markedly attenuated by nifedipine. These findings support evidence that the vasoconstrictor responses due to activation of alpha1-adrenoceptors are initiated by mobilization of intracellular pools of Ca(2+) whereas they are sustained by opening of Ca(2+)VERSUS-channels. These findings also suggest that PHMBA diminishes the vasoconstrictor effects of phenylephrine by chelation of thiol residues in alpha1-adrenoceptors rather than by blockade of Ca(2+)VERSUS-channels, and that chelation of these thiol residues prevents agonist occupation and/or activation of these receptors and subsequent mobilization of intracellular pools of Ca(2+).

Diversity of atrial local Ca2+ signalling: evidence from 2‐D confocal imaging in Ca2+‐buffered rat atrial myocytes

Atrial myocytes, lacking t-tubules, have two functionally separate groups of ryanodine receptors (RyRs): those at the periphery colocalized with dihydropyridine receptors (DHPRs), and those at the cell interior not associated with DHPRs. We have previously shown that the Ca(2+) current (I(Ca))-gated central Ca(2+) release has a fast component that is followed by a slower and delayed rising phase. The mechanisms that regulate the central Ca(2+) releases remain poorly understood. The fast central release component is highly resistant to dialysed Ca(2+) buffers, while the slower, delayed component is completely suppressed by such exogenous buffers. Here we used dialysis of Ca(2+) buffers (EGTA) into voltage-clamped rat atrial myocytes to isolate the fast component of central Ca(2+) release and examine its properties using rapid (240 Hz) two-dimensional confocal Ca(2+) imaging. We found two populations of rat atrial myocytes with respect to the ratio of central to peripheral Ca(2+) release (R(c/p)). In one population ('group 1', approximately 60% of cells), R(c/p) converged on 0.2, while in another population ('group 2', approximately 40%), R(c/p) had a Gaussian distribution with a mean value of 0.625. The fast central release component of group 2 cells appeared to result from in-focus Ca(2+) sparks on activation of I(Ca). In group 1 cells intracellular membranes associated with t-tubular structures were never seen using short exposures to membrane dyes. In most of the group 2 cells, a faint intracellular membrane staining was observed. Quantification of caffeine-releasable Ca(2+) pools consistently showed larger central Ca(2+) stores in group 2 and larger peripheral stores in group 1 cells. The R(c/p) was larger at more positive and negative voltages in group 1 cells. In contrast, in group 2 cells, the R(c/p) was constant at all voltages. In group 1 cells the gain of peripheral Ca(2+) release sites (Delta[Ca(2+)]/I(Ca)) was larger at -30 than at +20 mV, but significantly dampened at the central sites. On the other hand, the gains of peripheral and central Ca(2+) releases in group 2 cells showed no voltage dependence. Surprisingly, the voltage dependence of the fast central release component was bell-shaped and similar to that of I(Ca) in both cell groups. Removal of extracellular Ca(2+) or application of Ni(2+) (5 mM) suppressed equally I(Ca) and Ca(2+) release from the central release sites at +60 mV. Depolarization to +100 mV, where I(Ca) is absent and the Na(+)-Ca(2+) exchanger (NCX) acts in reverse mode, did not trigger the fast central Ca(2+) releases in either group, but brief reduction of [Na(+)](o) to levels equivalent to [Na(+)](i) facilitated fast peripheral and central Ca(2+) releases in group 2 myocytes, but not in group 1 myocytes. In group 2 cells, long-lasting (> 1 min) exposures to caffeine (10 mM) or ryanodine (20 microM) significantly suppressed I(Ca)-triggered central and peripheral Ca(2+) releases. Our data suggest significant diversity of local Ca(2+) signalling in rat atrial myocytes. In one group, I(Ca)-triggered peripheral Ca(2+) release propagates into the interior triggering central Ca(2+) release with significant delay. In a second group of myocytes I(Ca) triggers a significant number of central sites as rapidly and effectively as the peripheral sites, thereby producing more synchronized Ca(2+) releases throughout the myocytes. The possible presence of vestigial t-tubules and larger Ca(2+) content of central sarcoplasmic reticulum (SR) in group 2 cells may be responsible for the rapid and strong activation of central release of Ca(2+) in this subset of atrial myocytes.

Intracellular Ca2+ Pools and Fluxes in Cardiac Muscle-Derived H9c2 Cells

Relevant Ca(2+) pools and fluxes in H9c2 cells have been studied using fluorescent indicators and Ca(2+)-mobilizing agents. Vasopressin produced a cytoplasmic Ca(2+) peak with half-maximal effective concentration of 6 nM, whereas thapsigargin-induced Ca(2+) increase showed half-maximal effect at 3 nM. Depolarization of the mitochondrial inner membrane by protonophore was also associated with an increase in cytoplasmic Ca(2+). Ionomycin induced a small and sustained depolarization, while thapsigargin had a small but transient effect. The thapsigargin-sensitive Ca(2+) pool was also sensitive to ionomycin, whereas the protonophore-sensitive Ca(2+) pool was not. The vasopressin-induced cytoplasmic Ca(2+) signal, which caused a reversible discharge of the sarco-endoplasmic reticulum Ca(2+) pool, was sensed as a mitochondrial Ca(2+) peak but was unaffected by the permeability transition pore inhibitor cyclosporin A. The mitochondrial Ca(2+) peak was affected by cyclosporin A when the Ca(2+) signal was induced by irreversible discharge of the intracellular Ca(2+) pool, i.e., adding thapsigargin. These observations indicate that the mitochondria interpret the cytoplasmic Ca(2+) signals generated in the reticular store.

De‐eutrophication of a nitrogen‐saturated Scots pine forest by prescribed litter‐raking

AbstractShort‐term (&lt;7 years) effects of prescribed litter‐raking on forest‐floor nutrient pools, stand nutrition, and seepage water chemistry were studied in an N‐saturated Scots pine (Pinus sylvestris L.) forest in Southern Germany subject to high atmospheric‐nitrogen deposition. The study was based on a comparison of plots with and without annual prescribed litter raking at three sites with different N‐deposition levels. Prescribed litter‐raking resulted in a considerable reduction of forest‐floor thickness and mass, as well as of forest‐floor C, N, P, K, Mg, and Ca pools. Furthermore, it induced a significant decrease of the foliar N content in current‐year needles of the pines and a more balanced nutritional status of the stand. Particularly on the site subject to the highest N deposition, but to a lesser degree also at the other sites, the mean NO$ _3^- $ concentration in the subsoil seepage water and the N export into the groundwater were substantially reduced on the litter‐raked plots. The results show that in N‐saturated Scots pine ecosystems prescribed litter‐raking on areas of limited size, which are used as sources of groundwater‐derived drinking water and/or serve as habitat for endangered plant species, is a quick and effective method to achieve a more balanced nutritional status of the trees and to reduce seepage‐water NO$ _3^- $ concentrations and N export into the groundwater. In terms of sustainable ecosystem nutrient management, the conversion of conifer monocultures into broadleaf‐rich mixed stands is the better, yet less immediately effective method to reduce the seepage‐water N export from conifer forests subject to high atmospheric‐N deposition.

Acidic mist reduces foliar membrane-associated calcium and impairs stomatal responsiveness in red spruce

Acidic deposition can leach essential pools of calcium (Ca) directly from plant foliage. Because of the central role of Ca in environmental signal transduction, disruptions of labile foliar Ca pools could impair physiological responses to a variety of environmental stimuli and stressors. We investigated the possibility that acidic mist-induced depletion of membrane-associated Ca (mCa), which is one form of labile Ca, may alter stomatal responsiveness to water stress, a process known to include Ca in signal transduction cascades. Red spruce (Picea rubens Sarg.) seedlings were exposed to either pH 3.0 or pH 5.0 mist treatments for one growing season. Foliar nutrition was assessed following treatments, and declines in stomatal conductance and net photosynthesis were measured on current-year shoots following stem excision. Seedlings exposed to pH 3.0 acidic mist treatments had reduced mCa relative to the pH 5.0 treated seedlings. Seedlings subjected to the pH 3.0 acidic mist treatment exhibited impaired stomatal functions, including a smaller maximum aperture, slower closure and an increased lag time between stomatal closure and photosynthetic decline following experimental water stress. Delayed stomatal closure could undermine desiccation avoidance mechanisms. Previous work has demonstrated that acidic mist treatments deplete mCa in red spruce and impair cold tolerance, with similar effects in other species. The results we present provide further evidence that acidic mist-induced mCa depletion may cause disruption of a broad range of plant stress responses.

Mechanism of cGMP-mediated protection in a cellular model of myocardial reperfusion injury

Reperfusion injury of the myocardium is characterised by development of cardiomyocyte hypercontracture. Previous studies have shown that cGMP-mediated stimuli protect against reperfusion injury, but the cellular mechanism is still unknown. To simulate ischemia/reperfusion, adult rat cardiomyocytes were incubated anoxically (pH(o) 6.4) and then reoxygenated (pH(o) 7.4). Cytosolic calcium [Ca(2+)](i) (fura-2 ratio), pH(i) (BCECF ratio), cell length, and phospholamban phosphorylation were analysed. Under simulated ischemia cardiomyocytes develop [Ca(2+)](i) overload. When reoxygenated they rapidly undergo hypercontracture, triggered by oscillations of [Ca(2+)](i). We investigated whether cGMP-mediated stimuli can modulate [Ca(2+)](i) or pH(i) recovery and whether this contributes to their protective effect. Membrane-permeable cGMP analogues, 8-bromo-cGMP (1 mmol/L) or 8-pCPT-cGMP (10 micrommol/L), or a receptor-mediated activator of particulate guanylyl cyclase, urodilatin (1 micromol/L), were applied. The investigated stimuli protect against reoxygenation-induced hypercontracture (cell length as percent of end-ischemic length; control: 68+/-1.6; 8-bromo-cGMP: 88+/-1.5*; 8-pCPT-cGMP: 84+/-2.9*; urodilatin: 87+/-1.1*; n=24; *p<0.05). Recovery from [Ca(2+)](i) overload after 2 min reoxygenation [fura-2 ratio (a.u.); control: 1.43+/-0.15; 8-bromo-cGMP: 1.86+/-0.15*; 8-pCPT-cGMP: 1.92+/-0.19*; urodilatin: 1.93+/-0.24*; n=25; *p<0.05] was accelerated, and the frequency of [Ca(2+)](i) oscillations (min(-1)) was significantly reduced (control: 49+/-5.0 min(-1); 8-bromo-cGMP: 18+/-3.5* min(-1); 8-pCPT-cGMP: 18+/-4.5* min(-1); urodilatin: 16+/-4.1* min(-1); n=24; *p<0.05). cGMP-mediated stimuli increased sarcoplasmic Ca(2+) sequestration (caffeine-releasable Ca(2+) pool: 2-3 fold increase vs. control). Inhibition of sarcoplasmic Ca(2+)-ATPase (SERCA) by thapsigargin (150 nmol/L) or of protein kinase G with KT-5823 (1 micromol/L) abolished the effect of these stimuli on [Ca(2+)](i) recovery. The investigated stimuli significantly enhanced phospholamban phosphorylation. We conclude that cGMP-dependent signals activate SERCA via a protein kinase G-dependent phosphorylation of phospholamban. The increase in SERCA activity seems to reduce peak [Ca(2+)](i) and [Ca(2+)](i) oscillation during reoxygenation and to attenuate the excessive activation of the contractile machinery that otherwise leads to the development of hypercontracture.

Ca2+ Signaling in HEK-293 and Skeletal Muscle Cells Expressing Recombinant Ryanodine Receptors Harboring Malignant Hyperthermia and Central Core Disease Mutations

Malignant hyperthermia (MH) and central core disease (CCD) are caused by mutations in the RYR1 gene encoding the skeletal muscle isoform of the ryanodine receptor (RyR1), a homotetrameric Ca(2+) release channel. Rabbit RyR1 mutant cDNAs carrying mutations corresponding to those in human RyR1 that cause MH and CCD were expressed in HEK-293 cells, which do not have endogenous RyR, and in primary cultures of rat skeletal muscle, which express rat RyR1. Analysis of intracellular Ca(2+) pools was performed using aequorin probes targeted to the lumen of the endo/sarcoplasmic reticulum (ER/SR), to the mitochondrial matrix, or to the cytosol. Mutations associated with MH caused alterations in intracellular Ca(2+) homeostasis different from those associated with CCD. Measurements of luminal ER/SR Ca(2+) revealed that the mutations generated leaky channels in all cases, but the leak was particularly pronounced in CCD mutants. Cytosolic and mitochondrial Ca(2+) transients induced by caffeine stimulation were drastically augmented in the MH mutant, slightly reduced in one CCD mutant (Y523S) and completely abolished in another (I4898T). The results suggest that local Ca(2+) derangements of different degrees account for the specific cellular phenotypes of the two disorders.

New Insights into Calcium Depletion in Northeastern Forests

Abstract Acid rain and repeated harvests reduce the amount of calcium available for forest growth. Ecosystem budgets suggest that the exchangeable Ca pool is insufficient for forest regeneration, yet young stands appear to mobilize more than enough Ca from the soil to meet their needs. Extractions of soil parent materials indicate that apatite provides a previously unappreciated source of plant-available Ca for forests in New Hampshire, upstate New York, and Maine with granitoid parent materials. The threat of Ca depletion may not be as grave as previously predicted on these soil types. In contrast, apatite is not important at sites in Pennsylvania with sedimentary parent materials.

Assessment of calcium status in Maine forests: review and future projection

Forest harvesting and acidic deposition can cause substantial decreases in the calcium (Ca) inventory of forest soils if such losses are not replenished through mineral weathering, atmospheric deposition, or fertilization. The net balance between losses and gains defines the forest Ca status. Site-specific studies have measured Ca pools and fluxes in Maine forests, but no synthesis has been published. In this paper, I review the literature on forest Ca and assess the current status and potential future trends. Forest soils in Maine are currently at lesser risk of Ca depletion compared with many forest soils in the central and southeastern United States, because levels of acidic deposition and rates of Ca accumulation in trees are lower in Maine. The rate of Ca accumulation in trees is reduced in Maine as a result of lower growth rates and a higher proportion of conifer trees that require less Ca than hardwoods. However, field-scale biogeochemical studies in Maine and New Hampshire, and regional estimates of harvest removals and soil inventories coupled with low weathering estimates, indicate that Ca depletion is a realistic concern in Maine. The synthesis of site-specific and regional data for Maine in conjunction with the depletion measured directly in surrounding areas indicates that the Ca status of many forest soils in Maine is likely declining. Ca status could decrease further in the future if forest growth rates increase in response to climate trends and recovery from insect-induced mortality and excessive harvesting in recent years. Proposed climate change induced reductions in spruce and fir and increases in hardwoods would also increase the risk of Ca depletion.

Ryanodine receptor‐operated activation of TRP‐like channels can trigger critical Ca2+signaling events in pancreatic β‐cells

There is little information available concerning the link between the ryanodine (RY) receptors and the downstream Ca(2+) signaling events in beta-cells. In fura-2 loaded INS-1E cells, activation of RY receptors by 9-methyl 5,7-dibromoeudistomin D (MBED) caused a rapid rise of [Ca(2+)]i followed by a plateau and repetitive [Ca(2+)]i spikes on the plateau. The [Ca(2+)]i plateau was abolished by omission of extracellular Ca(2+) and by SKF 96365. In the presence of SKF 96365, MBED produced a transient increase of [Ca(2+)]i, which was abolished by thapsigargin. Activation of RY receptors caused Ca(2+) entry even when the ER Ca(2+) pool was depleted by thapsigargin. The [Ca(2+)]i plateau was not inhibited by nimodipine or ruthenium red, but was inhibited by membrane depolarization, La(3+), Gd(3+), niflumic acid, and 2-aminoethoxydiphenyl borate, agents that inhibit the transient receptor potential channels. The [Ca(2+)]i spikes were inhibited by nimodipine and ryanodine, indicating that they were due to Ca(2+) influx through the voltage-gated Ca(2+) channels and Ca(2+)-induced Ca(2+) release (CICR). Activation of RY receptors depolarized membrane potential as measured by patch clamp. Thus, activation of RY receptors leads to coherent changes in Ca(2+) signaling, which includes activation of TRP-like channels, membrane depolarization, activation of the voltage-gated Ca(2+) channels and CICR.

Hydrogen peroxide-induced extracellular signal-regulated kinase activation in cultured feline ileal smooth muscle cells.

H(2)O(2) has been shown to act as a signaling molecule involved in many cellular functions such as apoptosis and proliferation. In the present study, we characterized the effects of H(2)O(2) on the activation of mitogen-activated protein (MAP) kinases and examined the factors involved in the process of extracellular signal-regulated kinase (ERK) activation by H(2)O(2) in ileal smooth muscle cells (ISMC). ISMC were cultured and exposed to H(2)O(2). Western blot analysis was performed with phosphospecific MAP kinase antibodies. Potent activation of ERK and moderate activation of stress-activated protein kinase/c-Jun NH(2)-terminal kinase occurred within 30 min of 1 mM H(2)O(2) treatment. However, p38 MAP kinase was not activated by H(2)O(2). The activation of ERK by H(2)O(2) was reduced by the mitogen-activated/ERK-activating kinase inhibitor PD98059 [2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one], Ras inhibitor S-farnesylthiosalicylic acid, removal of extracellular Ca(2+), depletion of the intracellular Ca(2+) pool by thapsigargin, or pretreatment of ISMC with the calmodulin antagonist W-7. Also, H(2)O(2)-induced ERK activation was attenuated by a receptor tyrosine kinase inhibitor, tyrphostin 51, but not by down-regulation of protein kinase C (PKC) with phorbol 12-myristate 13-acetate or by a PKC inhibitor, GF109203X [3-[1-(dimethylaminopropyl)indol-3-yl]-4-(indol-3-yl)maleimide hydrochloride]. Growth factor receptor antagonist suramin pretreatment inhibited H(2)O(2)-induced ERK activation, highlighting a role for growth factor receptors in this activation. Furthermore, the ERK activation by H(2)O(2) was blocked by pretreatment with either N-acetyl-cysteine, o-phenanthroline, or mannitol indicating that metal-catalyzed free radical formation may mediate the initiation of signal transduction by H(2)O(2). These data suggest that short-term stimulation with H(2)O(2) activates the signaling pathways of cell mitogenic effects which are thought to be a protective response against intestinal oxidative stress.

Two Pathways for Store-mediated Calcium Entry Differentially Dependent on the Actin Cytoskeleton in Human Platelets

A major pathway for stimulated Ca(2+) entry in non-excitable cells is activated following depletion of intracellular Ca(2+) stores. Secretion-like coupling between elements in the plasma membrane (PM) and Ca(2+) stores has been proposed as the most likely mechanism to activate this store-mediated Ca(2+) entry (SMCE) in several cell types. Here we identify two mechanisms for SMCE in human platelets activated by depletion of two independent Ca(2+) pools, which are differentially modulated by the actin cytoskeleton. Ca(2+) entry induced by depletion of a 2,5-di-(tert-butyl)-1,4-hydroquinone (TBHQ)-sensitive pool is increased by disassembly of the actin cytoskeleton and that induced by a TBHQ-insensitive pool is reduced. Stabilization of the actin cytoskeleton prevented Ca(2+) entry by both mechanisms. We propose that the membrane-associated actin network prevents constitutive Ca(2+) entry via both pathways. Reorganization of the actin cytoskeleton permits the activation of Ca(2+) entry via both mechanisms, but only SMCE activated by the TBHQ-insensitive pool requires new actin polymerization, which may support membrane trafficking toward the PM.

Somatic and axonal effects of ATP via P2X2 but not P2X7 receptors in rat thoracolumbar sympathetic neurones.

Excitatory ATP responses in rat cultured thoracolumbar sympathetic neurones are mediated by somatic P2X(2) receptors. The present study investigated a possible role of axonal P2X(2) as well as P2X(7) receptors on the same preparation. Confocal laser scanning microscopy demonstrated P2X(2) and P2X(7) immunoreactivity along the axons as well as P2X(7) immunoreactivity surrounding the cell nuclei. P2X(7) mRNA expression was detected in individual neurones using a single-cell RT-PCR approach. Adenosine triphosphate (ATP) caused a significant increase in axonal Ca(2+) concentration which was dependent on external Ca(2+) but insensitive to depletion of the cellular Ca(2+) pools by cyclopiazonic acid. Pyridoxal-phosphate-6-azophenyl-2',4'-disulfonate (PPADS; 30 micro m) virtually abolished the ATP response, whereas brilliant blue G (0.1 micro m), a selective P2X(7) receptor antagonist, had no effect. Dibenzoyl-ATP (BzATP; 100 micro m) induced a much smaller increase in axonal [Ca(2+)] concentration than ATP at equimolar concentrations. The response to BzATP was distinctly reduced by PPADS but not by brilliant blue G. The overall pharmacological profile of the axonal P2X receptors resembled closely that of the somatic P2X(2) receptors. In conclusion, the present data suggest the occurrence of axonal excitatory P2X(2) receptors in thoracolumbar sympathetic neurones. However, the functional significance of axonal and (peri)-nuclear P2X(7) receptors has still to be proven.