Inorganic Osmolytes Research Articles

Screening of tomato genotypes for salinity tolerance based on early growth attributes and leaf inorganic osmolytes

ABSTRACTThe experiment containing three replicates of completely randomized factorial treatments was conducted in a glasshouse under controlled conditions with three simulated soil salinity levels (control, 10 and 15 dS m−1). Morpho-physiological traits (i.e. lengths, fresh weights and dry weights of root and shoot, number of leaves, root/shoot ratio, shoot Na+ accumulation, K+/Na+ ratio, Ca2+/Na+ ratio, membrane stability index, lycopene contents, chlorophyll-a and -b) were recorded to determine mechanism of salt tolerance of tomato at seedling stage. Principal component analysis (PCA) was used to express a three-way interaction of genotype × salinity level × traits that scattered the 25 tomato genotypes based on their morpho-physiological response to different NaCl levels. The negative association of Na+ with all other traits except root/shoot ratio and the morpho-physiological response trend of genotypes exposed that probable mechanism of salt tolerance was initially Na+ exclusion by abscising older leaves to have younger physiologically energetic, and lastly a higher activity of plants for root development to sustain them in saline soil. PCA three-way biplot efficiently recognized ANAHU, LA-2821, LO-2752, LO-2707, PB-017909, LO-2831-23 and 017860 as salt tolerant genotypes. On the other hand, ZARNITZA, GLACIER, LO-2692, LO-2576, BL-1079, 006233, 006232, 017856, NUTYT-701 and NAGINA were found to be salt susceptible.

Plant Growth is Highly Associated with the Concentration of Organic and Inorganic Osmolytes, Antioxidant Activities in Salt Stress Tomato (Lycopersicon esculentum) Plants

Plant Growth is Highly Associated with the Concentration of Organic and Inorganic Osmolytes, Antioxidant Activities in Salt Stress Tomato (Lycopersicon esculentum) Plants

Eco-physiological adaptations of Panicum antidotale to hyperosmotic salinity: Water and ion relations and anti-oxidant feedback

Threshold of salt resistance of plants is determined by their response to osmotic and ionic stress (primary constraints) imposed upon them. However, recent reports emphasize the importance of secondary constraints like oxidative stress. The aim of this study was to determine the effect of salinity on growth, mineral nutrition, water relations, compatible solutes, and the antioxidant system in Panicum antidotale.Five levels of salinity (0, 125, 250, 375 and 500mM NaCl) were applied using a quick check system in a fully randomized greenhouse study. Plant growth parameters, water relations, organic (proline and soluble sugars), inorganic osmolytes (Na+, K+, Ca++and Mg++), and macronutrients such as carbon or nitrogen were measured beside the activities of the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APx) and glutathione reductase (GR) and non-enzymatic antioxidant metabolites (oxidized and reduced ascorbate).Sodium chloride concentrations above 125mM substantially inhibited growth. This inhibition was attributed to high energy costs needed for osmotic adjustment, ion compartmentalization, synthesis of organic osmolytes (such as proline and sugars), ROS scavenging and the maintenance of ionic homeostasis. The plants resisted against oxidative stress by increasing activities of antioxidant enzymes such as SOD, APx, GR and CAT and elevating levels of oxidized and reduced ascorbate (DAsA and AsA) at higher salinity. They also maintained a low redox ratio of ascorbate/dehydro-ascorbate and therefore a high capacity to manage oxidative stress. Thus P. antidotale is capable of managing ROS stress at high salinity and therefore can be sustainably grown as a fodder crop in saline arid regions.

The blood biochemistry of overwintering diamondback terrapins (Malaclemys terrapin)

Estuarine ectothermic vertebrates, such as diamondback terrapins (Malaclemys terrapin, Schwartz 1955), inhabit a dynamic environment, and many aspects of their biology reflect their ability to withstand and respond to these environmental challenges. The physiological adjustments necessary to maintain water and salt balance and the metabolic adjustments that accompany seasonal changes in activity and behavior have not been well-characterized for overwintering terrapins under field conditions. To investigate terrapin osmotic and metabolic physiology during winter when activity levels are depressed, we obtained repeat blood samples from 10 radio-tagged female terrapins maintained in a semi-natural, open-air salt marsh enclosure in southeastern North Carolina, USA. From November 2011 to April 2012, we measured monthly plasma osmolality, plasma concentrations of inorganic osmolytes (Na+, K+, Cl−), and protein catabolic indices (urea and uric acid), as well as monthly plasma concentrations of total Ca2+, lactate, and glucose as metabolic indices. We used linear mixed models to determine the best predictors of blood chemistry, where time (i.e., day) and environmental variables were fixed factors and individual terrapins were random factors. Day was a poor predictor of blood chemistry concentrations, indicating that the progression of winter did not elicit corresponding changes in biochemical indices as documented in other semi-aquatic turtles exposed to more severe winter and/or laboratory conditions. Carapace temperature was the most common predictor of blood chemistry concentrations in all models, underscoring its relative influence on physiology. In contrast to previous laboratory-based studies on the overwinter physiology of terrapins, our study demonstrates that terrapins in their natural environment are able to maintain biochemical homeostasis throughout winter. The use of evasive behavioral strategies may be an important factor for terrapins to reduce the passive exchange of water and salts with the estuarine environment.

Exogenous 24-Epibrassinolide Elevates the Salt Tolerance Potential of Pea (Pisum sativum L.) by Improving Osmotic Adjustment Capacity and Leaf Water Relations

Salinity, as a wide spread problem around the globe, can drastically limits the crop growth and productivity. Therefore, there is a growing need to mitigate the drastic effects of this abiotic stress through the use some sustaiable short-term or long term approaches. An investigation was carried out to assess the potential role of 24-epibrassinolide (EBL) in mitigating the drastic effects of salt stress on growth, water relations, osmolytes, and ions accumulation of salt-tolerant and salt-sensitive pea genotypes. EBL treatment significantly altered the leaf water status and accumulation of osmolytes and various ions in salt-stressed [with sodium chloride (+NaCl)] plants. The plants of both salt-tolerant and salt-sensitive genotypes, subjected to +NaCl conditions had a significant reduction in plant fresh and dry weights, internodal distance, leaf area (LA), leaf water potential (w), leaf osmotic potential (o), leaf turgor potential (t), relative water content (RWC), number of seeds pod−1, seed weight, root/leaf inorganic osmolytes (Ca2+, K+ and Mg2+), while increased the levels of root/leaf proline, glycinebetaine, total free amino acids, total soluble sugars, sodium (Na+), and chloride (Cl−). There was non-significant effect of EBL on number of pods plant−1 both under saline and non-saline conditions. However, the foliar application of EBL significantly alleviated the deleterious effects of salinity by improving the plant biomass, water relations and concentration of organic/inorganic osmolytes and yield attributes. However, EBL minimized the drastic effects of salt stress by limiting the root/leaf Na+ and Cl− contents. It can be concluded that EBL efficiently alleviated the salinity-induced drastic effects by improving the water status of plant tissues, which led to salt dilution within plant tissues, thus minimized the ion-toxicity, ultimately elevated the plant growth by minimizing the toxic action of Na+ and Cl−. Secondly, EBL augmented the salt-tolerance capacity of tested pea genotypes by enhancing their osmotic adjustment potential, in terms of high accumulation of organic and inorganic osmolytes within leaf and root tissues.

Ion homeostasis in a salt-secreting halophytic grass.

Salinity adversely affects plant growth and development, and disturbs intracellular ion homeostasis, resulting in cellular toxicity. Plants that tolerate salinity, halophytes, do so by manifesting numerous physiological and biochemical processes in coordination to alleviate cellular ionic imbalance. The present study was undertaken to analyse the salt tolerance mechanism in Aeluropus lagopoides (L.) trin. Ex Thw. (Poaceae) at both physiological and molecular levels. Plants secreted salt from glands, which eventually produced pristine salt crystals on leaves and leaf sheaths. The rate of salt secretion increased with increasing salt concentration in the growth medium. Osmotic adjustment was mainly achieved by inorganic osmolytes (Na(+)) and at 100 mM NaCl no change was observed in organic osmolytes in comparison to control plants. At 300 mM NaCl and with 150 mM NaCl + 150 mM KCl, the concentration of proline, soluble sugars and amino acids was significantly increased. Transcript profiling of transporter genes revealed differential spatial and temporal expressions in both shoot and root tissues in a manner synchronized towards maintaining ion homeostasis. In shoots, AlHKT2;1 transcript up-regulation was observed at 12 and 24 h in all the treatments, whereas in roots, maximum induction was observed at 48 h with K(+) starvation. The HAK transcript was relatively abundant in shoot tissue with all the treatments. The plasma membrane Na(+)/H(+) antiporter, SOS1, and tonoplast Na(+)/H(+) antiporter, NHX1, were found to be significantly up-regulated in shoot tissue. Our data demonstrate that AlHKT2;1, HAK, SOS1, NHX1 and V-ATPase genes play a pivotal role in regulating the ion homeostasis in A. lagopoides.

Linking osmotic adjustment and stomatal characteristics with salinity stress tolerance in contrasting barley accessions.

Salinity tolerance is a complex trait - both physiologically and genetically - and the issue of which mechanism or trait has bigger contribution towards the overall plant performance is still hotly discussed in the literature. In this work, a broad range of barley (Hordeum vulgare L. and Hordeum spontaneum L.) genotypes contrasting in salinity stress tolerance were used to investigate the causal link between plant stomatal characteristics, tissue ion relations, and salinity tolerance. In total, 46 genotypes (including two wild barleys) were grown under glasshouse conditions and exposed to moderate salinity stress (200mM NaCl) for 5 weeks. The overall salinity tolerance correlated positively with stomata density, leaf K+ concentration and the relative contribution of inorganic ions towards osmotic adjustment in the shoot. At the same time, no correlation between salinity tolerance and stomatal conductance or leaf Na+ content in the shoot was found. Taken together, these results indicate the importance of increasing stomata density as an adaptive tool to optimise efficiency of CO2 assimilation under moderate saline conditions, as well as benefits of the predominant use of inorganic osmolytes for osmotic adjustment in barley. Another finding of note was that wild barleys showed rather different strategies dealing with salinity, as compared with cultivated varieties.

Oxidative stress protection and stomatal patterning as components of salinity tolerance mechanism in quinoa (Chenopodium quinoa)

Two components of salinity stress are a reduction in water availability to plants and the formation of reactive oxygen species. In this work, we have used quinoa (Chenopodium quinoa), a dicotyledonous C3 halophyte species displaying optimal growth at approximately 150 mM NaCl, to study mechanisms by which halophytes cope with the afore-mentioned components of salt stress. The relative contribution of organic and inorganic osmolytes in leaves of different physiological ages (e.g. positions on the stem) was quantified and linked with the osmoprotective function of organic osmolytes. We show that the extent of the oxidative stress (UV-B irradiation) damage to photosynthetic machinery in young leaves is much less when compared with old leaves, and attribute this difference to the difference in the size of the organic osmolyte pool (1.5-fold difference under control conditions; sixfold difference in plants grown at 400 mM NaCl). Consistent with this, salt-grown plants showed higher Fv/Fm values compared with control plants after UV-B exposure. Exogenous application of physiologically relevant concentrations of glycine betaine substantially mitigated oxidative stress damage to PSII, in a dose-dependent manner. We also show that salt-grown plants showed a significant (approximately 30%) reduction in stomatal density observed in all leaves. It is concluded that accumulation of organic osmolytes plays a dual role providing, in addition to osmotic adjustment, protection of photosynthetic machinery against oxidative stress in developing leaves. It is also suggested that salinity-induced reduction in stomatal density represents a fundamental mechanism by which plants optimize water use efficiency under saline conditions.

Vacuolar Transporters in Their Physiological Context

Vacuoles in vegetative tissues allow the plant surface to expand by accumulating energetically cheap inorganic osmolytes, and thereby optimize the plant for absorption of sunlight and production of energy by photosynthesis. Some specialized cells, such as guard cells and pulvini motor cells, exhibit rapid volume changes. These changes require the rapid release and uptake of ions and water by the vacuole and are a prerequisite for plant survival. Furthermore, seed vacuoles are important storage units for the nutrients required for early plant development. All of these fundamental processes rely on numerous vacuolar transporters. During the past 15 years, the transporters implicated in most aspects of vacuolar function have been identified and characterized. Vacuolar transporters appear to be integrated into a regulatory network that controls plant metabolism. However, little is known about the mode of action of these fundamental processes, and deciphering the underlying mechanisms remains a challenge for the future.

Analysis of constituents for phenotyping drought tolerance in crop improvement.

Investigators now have a wide range of analytical tools to use in measuring metabolites, proteins and transcripts in plant tissues. These tools have the potential to assist genetic studies that seek to phenotype genetic lines for heritable traits that contribute to drought tolerance. To be useful for crop breeding, hundreds or thousands of genetic lines must be assessed. This review considers the utility of assaying certain constituents with roles in drought tolerance for phenotyping genotypes. Abscisic acid (ABA), organic and inorganic osmolytes, compatible solutes, and late embryogenesis abundant proteins, are considered. Confounding effects that require appropriate tissue and timing specificity, and the need for high-throughput and analytical cost efficiency are discussed. With future advances in analytical methods and the value of analyzing constituents that provide information on the underlying mechanisms of drought tolerance, these approaches are expected to contribute to development crops with improved drought tolerance.

Ion transport and osmotic adjustment in plants and bacteria.

Plants and bacteria respond to hyperosmotic stress by an increase in intracellular osmolality, adjusting their cell turgor to altered growth conditions. This can be achieved either by increased uptake or de novo synthesis of a variety of organic osmolytes (so-called 'compatible solutes'), or by controlling fluxes of ions across cellular membranes. The relative contributions of each of these mechanisms have been debated in literature for many years and remain unresolved. This paper summarises all the arguments and reopens a discussion on the efficiency and strategies of osmotic adjustment in plants and bacteria. We show that the bulk of osmotic adjustment in both plants and bacteria is achieved by increased accumulation of inorganic osmolytes such as K+, Na+ and Cl-. This is applicable to both halophyte and glycophyte species. At the same time, de novo synthesis of compatible solutes is an energetically expensive and slow option and can be used only for the fine adjustment of the cell osmotic potential. The most likely role the organic osmolytes play in osmotic adjustment is in osmoprotection of key membrane transport proteins and reactive oxygen species (ROS) scavenging. The specific mechanisms by which compatible solutes regulate activity of ion transporters remain elusive and require more thorough investigation. It is concluded that creating transgenic species with increased levels of organic osmolytes by itself is counterproductive due to high yield penalties; all these attempts should be complemented by a concurrent increase in the accumulation of inorganic ions directly used for osmotic adjustment.

Receptor regulation of osmolyte homeostasis in neural cells

The capacity of cells to correct their volume in response to hyposmotic stress via the efflux of inorganic and organic osmolytes is well documented. However, the ability of cell-surface receptors, in particular G-protein-coupled receptors (GPCRs), to regulate this homeostatic mechanism has received much less attention. Mechanisms that underlie the regulation of cell volume are of particular importance to cells in the central nervous system because of the physical restrictions of the skull and the adverse impact that even small increases in cell volume can have on their function. Increases in brain volume are seen in hyponatraemia, which can arise from a variety of aetiologies and is the most frequently diagnosed electrolyte disorder in clinical practice. In this review we summarize recent evidence that the activation of GPCRs facilitates the volume-dependent efflux of osmolytes from neural cells and permits them to more efficiently respond to small, physiologically relevant, reductions in osmolarity. The characteristics of receptor-regulated osmolyte efflux, the signalling pathways involved and the physiological significance of receptor activation are discussed. In addition, we propose that GPCRs may also regulate the re-uptake of osmolytes into neural cells, but that the influx of organic and inorganic osmolytes is differentially regulated. The ability of neural cells to closely regulate osmolyte homeostasis through receptor-mediated alterations in both efflux and influx mechanisms may explain, in part at least, why the brain selectively retains its complement of inorganic osmolytes during chronic hyponatraemia, whereas its organic osmolytes are depleted.

Seed germination, seedling survival, and physiological response of sunflowers under saline and alkaline conditions

Salinization and alkalization of soil are widespread environmental problem and the alkali stress is more destructive than the effects caused by salt stress. To compare the mechanism of salt and alkali stresses, a sunflower variety (Helianthus annuus L. cv. Baikuiza 6) was tested under saline or alkaline conditions by mixing two neutral salts (NaCl and Na2SO4) or two alkaline salts (NaHCO3 and Na2CO3). The results showed that saline conditions differed greatly from alkaline conditions in their threshold intensities where sunflower can germinate, survive and grow. Under saline conditions, the emergence time was delayed, and the emergence rate and seedling survival rate also decreased with increasing salinity. However, under alkaline conditions, the rate of seedling survival decreased sharply but the emergence time and emergence rate did not change. In addition, the damaging effects of alkali stress on growth and photosynthesis were more severe than those of saline. In shoots, the main inorganic osmolyte and cation was K+ rather than Na+; the primary organic osmolytes were organic acid and soluble sugar rather than proline. Organic acid, NO3 −, and Cl− (only under saline condition) were the main source of anion. In addition, the osmotic adjustment and ion balance differed among sunflower roots, stems, and leaves. In conclusion, saline and alkaline conditions are two different stress conditions and there are special responses to two stress conditions for sunflower.

Heteromeric AtKC1·AKT1 Channels in Arabidopsis Roots Facilitate Growth under K+-limiting Conditions

Plant growth and development is driven by osmotic processes. Potassium represents the major osmotically active cation in plants cells. The uptake of this inorganic osmolyte from the soil in Arabidopsis involves a root K(+) uptake module consisting of the two K(+) channel alpha-subunits, AKT1 and AtKC1. AKT1-mediated potassium absorption from K(+)-depleted soil was shown to depend on the calcium-sensing proteins CBL1/9 and their interacting kinase CIPK23. Here we show that upon activation by the CBL.CIPK complex in low external potassium homomeric AKT1 channels open at voltages positive of E(K), a condition resulting in cellular K(+) leakage. Although at submillimolar external potassium an intrinsic K(+) sensor reduces AKT1 channel cord conductance, loss of cytosolic potassium is not completely abolished under these conditions. Depending on channel activity and the actual potassium gradients, this channel-mediated K(+) loss results in impaired plant growth in the atkc1 mutant. Incorporation of the AtKC1 subunit into the channel complex, however, modulates the properties of the K(+) uptake module to prevent K(+) loss. Upon assembly of AKT1 and AtKC1, the activation threshold of the root inward rectifier voltage gate is shifted negative by approximately -70 mV. Additionally, the channel conductance gains a hypersensitive K(+) dependence. Together, these two processes appear to represent a safety strategy preventing K(+) loss through the uptake channels under physiological conditions. Similar growth retardation phenotypes of akt1 and atkc1 loss-of-function mutants in response to limiting K(+) supply further support such functional interdependence of AKT1 and AtKC1. Taken together, these findings suggest an essential role of AtKC1-like subunits for root K(+) uptake and K(+) homeostasis when plants experience conditions of K(+) limitation.

The Fastest Flights in Nature: High-Speed Spore Discharge Mechanisms among Fungi

BackgroundA variety of spore discharge processes have evolved among the fungi. Those with the longest ranges are powered by hydrostatic pressure and include “squirt guns” that are most common in the Ascomycota and Zygomycota. In these fungi, fluid-filled stalks that support single spores or spore-filled sporangia, or cells called asci that contain multiple spores, are pressurized by osmosis. Because spores are discharged at such high speeds, most of the information on launch processes from previous studies has been inferred from mathematical models and is subject to a number of errors.Methodology/Principal FindingsIn this study, we have used ultra-high-speed video cameras running at maximum frame rates of 250,000 fps to analyze the entire launch process in four species of fungi that grow on the dung of herbivores. For the first time we have direct measurements of launch speeds and empirical estimates of acceleration in these fungi. Launch speeds ranged from 2 to 25 m s−1 and corresponding accelerations of 20,000 to 180,000 g propelled spores over distances of up to 2.5 meters. In addition, quantitative spectroscopic methods were used to identify the organic and inorganic osmolytes responsible for generating the turgor pressures that drive spore discharge.Conclusions/SignificanceThe new video data allowed us to test different models for the effect of viscous drag and identify errors in the previous approaches to modeling spore motion. The spectroscopic data show that high speed spore discharge mechanisms in fungi are powered by the same levels of turgor pressure that are characteristic of fungal hyphae and do not require any special mechanisms of osmolyte accumulation.

Effects of osmolytes on hexokinase kinetics combined with macromolecular crowding: Test of the osmolyte compatibility hypothesis towards crowded systems

We investigated the effect of compatible and non-compatible osmolytes in combination with macromolecular crowding on the kinetics of yeast hexokinase. This was motivated by the fact that almost all studies concerning the osmolyte effects on enzyme activity have been performed in diluted buffer systems, which are far from the physiological conditions within cells, where the cytosol contains several hundred mg protein ml − 1 . Four organic (glycerol, betaine, TMAO and urea) and one inorganic (NaCl) osmolyte were tested. It was concluded that the effect of compatible osmolytes (glycerol, betaine and TMAO) on V max and K M was practically equivalent in pure buffer and in 200–250 mg BSA ml − 1 supporting the view that these small organic osmolytes do minimal perturbance on enzyme function in physiological solutions. The effect of urea on enzyme kinetics was not independent of protein concentration, since the presence of 250 mg BSA ml − 1 partly compensated the perturbing effect of urea. Even though the organic osmolytes glycerol, betaine and TMAO are generally considered compatible with enzyme function, especially glycerol did have a significant effect on hexokinase kinetics, decreasing both k cat, K M and k cat/ K M. The osmolytes decreased k cat/ K M in the order: NaCl > Urea > TMAO/glycerol > betaine. For the organic osmolytes this order correlates with the degree of exclusion from protein–water interfaces. Thus, the stronger the exclusion the weaker the perturbing effects on k cat/ K M.

Osmotic stress and muscle tissue volume response of a freshwater bivalve

The freshwater bivalve, Corbicula fluminea, when submitted to hyperosmotic solutions, behaves as a hyperosmoconformer; we have observed an increase in osmolality and ions in its extracellular fluid. Osmotic and ionic changes in its watery environment represent a challenge for the tissues of this mollusk. Thus we evaluated, in vitro, muscle tissue volume variations (based on wet weight change) under anisosmotic salines, as well the possible regulatory mechanisms involved in the processes. This tissue did not exhibit complete volume regulation under anisosmotic saline solutions, but showed less variation than would be predicted by Van't Hoff's law, and tissue volume remained essentially stable throughout 90 min of exposure. To minimize tissue swelling in hyposmotic situations, C. fluminea muscle mobilizes organic osmolytes (ninhydrin positive substances) and inorganic ions (K+ and Cl−). While under hyperosmotic stimulus, apparently only inorganic osmolytes (Na+ and Cl−) are mobilized by the tissue. Our results indicate ionic accumulation by the Na+–K+–2Cl− cotransporter and the Na+/H+ coupled to Cl−/HCO3− exchangers. Exposure of the muscle tissue to Ca2+-free anisosmotic saline did not result in a detectable inhibition of the mechanisms described above. The Ca2+ gradient that derives from the absence of this ion, even apparently enhances the regulatory mechanisms. These responses of this freshwater mollusk in hyperosmotic solutions, and the muscle tissue under anisosmotic (hypo and hyperosmotic) saline solutions, have not been previously characterized in the manner and approach as reported here. Specifically, we analyze both organic and inorganic osmolytes mobilized under hyposmotic stress, and can infer the participation of Na+ and Cl− pathways stimulated by hyperosmotic stress. From the perspective gained in this study, tissue volume responses may be used as models for toxicological investigations.

Feeding and osmoregulation: dual function of the marine teleost intestine

Experiments on Gulf toadfish Opsanus beta demonstrate how feeding impacts osmoregulation in the marine teleost intestine. A high Ca(2+) diet of pilchards Sardina pilchardus ([Ca(2+)]=404.2 mmol kg(-1)) was compared to a low Ca(2+) diet of common squid Loligo forbesi ([Ca(2+)]=1.3 mmol kg(-1)), as high [Ca(2+)] has been shown to stimulate intestinal anion exchange. Gastrointestinal fluids and blood plasma were collected over a time course from pre-feeding to 216 h post feeding. Following food intake, monovalent ions were largely absorbed across the intestinal epithelium, leaving a fluid rich in divalent ions, which have a lower osmotic coefficient and effectively reduce osmotic pressure in the lumen to allow for enhanced fluid absorption. Concentrations of Cl(-) and HCO (-)(3) in fluid along the gastrointestinal tract of fish fed both diets, particularly 1 and 2 days post-feeding, demonstrate that apical Cl(-)/HCO (-)(3) exchange plays a vital role in postprandial Cl(-) and water absorption. Postprandial acid-base balance disturbance as indicated by plasma alkalinization was limited or absent, indicating compensation for gastric acid secretion in this teleost fish. Plasma osmolality peaked 12 h post-feeding in toadfish fed squid, but was not accompanied by a significant increase in inorganic ion concentrations. Transient fluid secretion by the gastrointestinal tract was evident from reduced luminal Mg(2+) and SO (2-)(4) concentrations for 24-48 h post feeding. Discrepancy between the sum of inorganic osmolytes and measured osmotic pressure was attributed to organic osmolytes, which occurred at high concentrations in the stomach and anterior intestine for up to 24 h post feeding.

Leishmania donovani Amastigotes Mobilize Organic and Inorganic Osmolytes During Regulatory Volume Decrease

The protozoan parasite Leishmania donovani encounters large fluctuations in osmolality as it cycles between its insect vector and human host. The flagellated promastigote exhibits regulatory volume responses involving organic and inorganic osmolytes, but little is known about volume regulation in the clinically relevant amastigote that multiplies within the parasitophorous vacuoles of mammalian host cells. Using a combination of morphological, X-ray microanalytical, and biochemical approaches we determined that non-motile amastigotes respond to hypotonic stress with (1) an amino acid and l-alanine-mediated regulatory volume decrease, and (2) a parallel release of Na+, K+, P (presumably as negatively charged phosphates), and subsequently Cl- from cytoplasm and the cell as a whole. In addition P, Zn2+, and subsequently Ca2+ increase in acidocalcisomes as Cl- content declines in this compartment. This evidence is the first to document subcellular translocation of, and thus a potential role for, zinc in volume regulatory responses. These coordinated changes in organic and inorganic osmolytes demonstrate that amastigote subcellular compartments, particularly acidocalcisomes, function in maintaining ionic homeostasis in the response of Leishmania amastigotes to hypo-osmotic stress.

Ion channels and transporters involved in cell volume regulation and sensor mechanisms.

All animal cell types have an appropriate volume. Even under physiological conditions of constant extracellular osmolarity, cells must regulate their volume. Cell volume is subjected to alterations because of persistent physicochemical osmotic load resulting from Donnan-type colloid osmotic pressure and of cell activity-associated changes in intracellular osmolarity resulting from osmolyte transport and metabolism. The strategy adopted by animal cells for coping with volume regulation on osmotic perturbation is to activate transport pathways, including channels and transporters, mainly for inorganic osmolytes to drive water flow. Under normotonic conditions, cells undergo volume regulation by pump-mediated mechanisms. Under anisotonic conditions, volume regulation occurs by additional channel/transporter-mediated mechanisms. Cell volume regulation is also attained through adjustment of intracellular levels not only of inorganic but also of organic osmolytes with changing the expression of their transporters or regulation of metabolism. In cell volume regulation mechanism, several "volume sensors" are thought to be involved. A volume-sensitive Cl- channel has lately attracted considerable attention in this regard.