Potassium-selective channel in the red beet vacuolar membrane

In higher plants the vacuolar K(+)-selective (VK) channel was identified solely in guard cells. This patch-clamp study describes a 40 pS homologue of the VK channel in Beta vulgaris taproot vacuoles. This voltage-independent channel is activated by submicromolar Ca(2+), and is ideally selective for K(+) over Cl(-) and Na(+).     

Infection by Trypanosoma cruzi Enhances Anion Conductance in Rat Neonatal Ventricular Cardiomyocytes

Recent studies on malaria-infected erythrocytes have shown increased anion channel activity in the host cell membrane, increasing the exchange of solutes between the cytoplasm and exterior. In the present work, we addressed the question of whether another intracellular protozoan parasite, Trypanosoma cruzi, alters membrane transport systems in the host cardiac cell. Neonatal rat cardiomyocytes were cultured and infected with T. cruzi in vitro. Ion currents were measured by patch-clamp technique in the whole-cell configuration. Two small-magnitude instantaneous anion currents, outward- and inward-rectifying, were recorded in all noninfected cardiomyocytes. In addition, ~10% of cardiomyocytes expressed a large anion-preferable, time-dependent current activated at positive membrane potentials. Hypotonic (230?mOsm) treatment resulted in the disappearance of the time-dependent current but provoked a dramatic increase of the instantaneous outward-rectifying one. Both instantaneous currents were suppressed by intracellular Mg(2+). T. cruzi infection did not provoke new anion currents in the host cells but caused an increase of the density of intrinsic swelling-activated outward current, up to twice in heavily infected cells. The occurrence of a time-dependent current dramatically increased in infected cells in the presence of Mg(2+) in the intracellular solution, from ~10 to ~80%, without a significant change of the current density. Our findings represent one further, besides the known Plasmodium falciparum, example of an intracellular parasite which upregulates the anionic currents expressed in the host cell.     

Palaeoecological significance of Barremian ammonite assemblages and facies variations from Southwest Mexico

Analyses of ammonite shell forms of two Barremian stratigraphic sections from Southwest Mexico consist of two well-defined morphotypes: (1) Small uncoiled, mostly leptoceratoid ancyloconic shells of the families Ancyloceratidae and Hamulinidae, and (2) middle-sized involute to moderately evolute oxycone to discocone shells of the family Pulchelliidae. Index taxa allow the recognition of standard ammonite biozones for the Barremian, which permit the relative dating of different processes that occurred through the water column in the environment of deposition. The vertical distribution of ammonite morphotypes and facies suggests changes of the palaeoceanographic and sedimentological conditions that prevailed in the area during Barremian time. Petrologic data, analyses of the organic carbon and carbonate contents of the rocks support the idea that oxygen-deficient bottom waters existed within a shallow marine, tectonically active area with little carbonate deposition during the early early Barremian (upper part of the Taveraidiscus hugii Zone through the base of the Nicklesia pulchella Zone). These conditions in the basin caused a proliferation of middle-water depth ammonites of Morphotype 1 but prevented the abundance of nektobenthic forms of Morphotype 2. Oxic conditions on a more calcareous and open normal marine environment seem to have been reestablished progressively during a transgressive episode from late early-early late Barremian (upper part of the Nicklesia pulchella Zone through the Gerhardtia sartousiana Zone). This environmental setting supported more facies dependent nektobenthic ammonites of Morphotype 2 to flourish within the basin.     

Asymmetric block of the plant vacuolar Ca2+-permeable channel by organic cations

In this work we have analysed the voltage-dependent block of the slow activating channel from red beet vacuoles by Tris, quaternary ammonium ions and the natural polyamines putrescine, spermidine and spermine. All these organic cations when applied from the cytosolic side blocked the channel by binding apparently deep (zδ values in the range of 0.65–1.35) within the pore. Tetraethylammonium ion did not pass the selectivity filter, whereas the cations with a smaller cross-section and Tris could pass across the entire pore, as evidenced by a relief of block at high positive voltages. Voltage dependence of the establishment of block from cytosolic side and of its relief was anomalously strong in the sense that the total charge moved across the pore for all blockers tested, with a notable exception of spermine, was in excess of their actual valence. This behaviour is consistent with the existence of multiple binding sites within a long pore, their simultaneous occupancy and interaction between different ions. In contrast, binding of blockers from the vacuolar (lumenal) side appears to follow a single-ion handling rule, with a common binding site for all amines located at approximately 30% of the electrical distance from the lumenal side. Received: 22 February 1999 / Revised version: 6 July 1999 / Accepted: 8 July 1999     

Mechanism of luminal Ca<Superscript>2+</Superscript> and Mg<Superscript>2+</Superscript> action on the vacuolar slowly activating channels

The non-selective slow vacuolar (SV) channel can dominate tonoplast conductance, making it necessary to tightly control its activity. Applying the patch-clamp technique to vacuoles from sugar beet (Beta vulgaris L.) taproots we studied the effect of divalent cations on the vacuolar side of the SV channel. Our results show that the SV channel has two independent binding sites for vacuolar divalent cations, (i) a less selective one, inside the channel pore, binding to which impedes channel conductance, and (ii) a Ca²⁺-selective one outside the membrane-spanning part of the channel protein, binding to which stabilizes the channels closed conformations. Vacuolar Ca²⁺ and Mg²⁺ almost indiscriminately blocked ion fluxes through the open channel pore, decreasing measured single-channel current amplitudes. This low-affinity block displays marked voltage dependence, characteristic of a permeable blocker. Vacuolar Ca²⁺—with a much higher affinity than Mg²⁺—slows down SV channel activation and shifts the voltage dependence to more (cytosol) positive potentials. A quantitative analysis results in a model that exactly describes the Ca²⁺-specific effects on the SV channel activation kinetics and voltage gating. According to this model, multiple (approximately three) divalent cations bind with a high affinity at the luminal interface of the membrane to the channel protein, favoring the occupancy of one of the SV channels closed states (C2). Transition to another closed state (C1) diminishes the effective number of bound cations, probably due to mutual repulsion, and channel opening is accompanied by a decrease of binding affinity. Hence, the open state (O) is destabilized with respect to the two closed states, C1 and C2, in the presence of Ca²⁺ at the vacuolar side. The specificity for Ca²⁺ compared to Mg²⁺ is explained in terms of different binding affinities for these cations. In this study we demonstrate that vacuolar Ca²⁺ is a crucial regulator to restrict SV channel activity to a physiologically meaningful range, which is less than 0.1% of maximum SV channel activity.Abbreviation SV Slow vacuolar     

Origin and Phylogeny of Microbes Living in Permanent Antarctic Lake Ice

Abstract The phylogenetic diversity of bacteria and cyanobacteria colonizing sediment particles in the permanent ice cover of an Antarctic lake was characterized by analyses of 16S rRNA genes amplified from environmental DNA. Samples of mineral particles were collected from a depth of 2.5 m in the 4-m-thick ice cover of Lake Bonney, McMurdo Dry Valleys, Antarctica. A rRNA gene clone library of 198 clones was made and characterized by sequencing and oligonucleotide probe hybridization. The library was dominated by representatives of the cyanobacteria, proteobacteria, and Planctomycetales, but also contained diverse clones representing many other microbial groups, including the Acidobacterium/Holophaga division, the Green Non-Sulfur division, and the Actinobacteria. Six oligonucleotide probes were made for the most abundant clades recovered in the library. To determine whether the ice microbial community might originate from wind dispersal of the algal mats found elsewhere in Taylor Valley, the probes were hybridized to 16S rDNAs amplified from three samples of terrestrial cyanobacterial mats collected at nearby sites, as well as to bacterial 16S rDNAs from the lake ice community. The results demonstrate the presence of a diverse microbial community dominated by cyanobacteria in the lake ice, and also show that the dominant members of the lake ice microbial community are found in terrestrial mats elsewhere in the area. The lake ice microbial community appears to be dominated by organisms that are not uniquely adapted to the lake ice ecosystem, but instead are species that originate elsewhere in the surrounding region and opportunistically colonize the unusual habitat provided by the sediments suspended in lake ice. Received: 16 August 1999; Accepted: 28 December 1999; Online Publication: 28 April 2000     

Conduction of Monovalent and Divalent Cations in the Slow Vacuolar Channel

The conduction properties of individual physiologically important cations Na+, K+, Mg2+, and Ca2+ were determined in the slowly activating (SV) channel of sugar beet vacuoles. Current-voltage relationships of the open channel were measured on excised tonoplast patches in a continuous manner by applying a +/-140 mV ramp-wave protocol. Applying KCl gradients of either direction across the patch we have determined that the relative Cl- to K+ permeability was < or =1%. Symmetrical increase of the concentration of tested cation caused an increase of the single channel conductance followed by saturation. Fitting of binding isotherms at zero voltage to the Michaelis-Menten equation resulted in values of maximal conductance of 300, 385, 18, and 13 pS, and of apparent dissociation constants of 64, 103, 0.04, and 0.08 mm for Na+, K+, Mg2+, and Ca2+, respectively. Deviations from the single-ion occupancy mechanism are documented, and alternative models of permeation are discussed. The magnitude of currents carried by divalent cations at low concentrations can be explained by an unrealistically wide (approximately 140 A) radius of the pore entrance. We propose instead a fixed negative charge in the pore vestibules, which concentrates the cations in their proximity. The conduction properties of the SV channel are compared with reported characteristics of voltage-dependent Ca2+-permeable channels, and consequences for a possible reduction of postulated multiplicity of Ca2+ pathways across the tonoplast are drawn.     

Factors controlling upper Jurassic ammonite assemblages in north-central Mexico

Lower Kimmeridgian to Lower Tithonian (Upper Jurassic) sections studied at Sierra de Palotes (Durango) and Sierra de Catorce (San Luis Potosí), Mexico, show low-energy deposits in which the composition of fossil macroinvertebrate assemblages, including megabenthos, reflects biostratinomic control. Monotonous siltstones provide continuous records of ammonite assemblages and reflect dominant deposition of shells in living areas; meanwhile, discontinues records were forced by episodic post-mortem transportation of shells, which was especially accentuated under storm influence. Rhythmic marly-silty limestones and marls illustrate a fossil record probably determined by minor transgressive-regressive pulses. The major changes in lithofacies are reflected by condensed silty and phosphatic mudstones deposited during significant floodings affecting areas under dominant terrigenous sedimentation. These changes determined more or less significant variations in the composition of fossil assemblages according to their relation to changing ecological conditions. However, shifting ecospaces exhibit no direct relationship to changes in lithofacies. Post-mortem transportation, operating in relation to both marine floodings and changes in the pattern of upper-water currents, was the main biostratinomic factor affecting the areal distribution of ammonite populations. Shell transportation and sedimentation rate controlled preservation and ultimately influenced diversity in recorded ammonite assemblages. The post-mortem behaviour (interpreted from shell structure and preservation), and therefore distribution, of ammonite shells points to shallow-water environments during the Kimmeridgian - Early Tithonian in areas (such as SE Durango and San Luís Potosí) close to the changing boundary between dominant carbonate and terrigenous sedimentation. No reworking affecting ammonite biostratigraphy has been identified in the sections studied.