Intrinsic Gating Properties of a Cloned G Protein-activated Inward Rectifier K^+ Channel

The voltage-, time-, and K^+-dependent properties of a G protein-activated inwardly rectifying K^+ channel (GIRK1/KGA/Kir3.1) cloned from rat atrium were studied in Xenopus oocytes under two-electrode voltage clamp. During maintained G protein activation and in the presence of high external K^+ (V_K = 0 mV), voltage jumps from V_K to negative membrane potentials activated inward GIRK1 K^+ currents with three distinct time-resolved current components. GIRK1 current activation consisted of an instantaneous component that was followed by two components with time constants T_f~50 ms and T_s~400 ms. These activation time constants were weakly voltage dependent, increasing approximately twofold with maximal hyperpolarization from V_K. Voltage-dependent GIRK1 availability, revealed by tail currents at -80 mV after long prepulses, was greatest at potentials negative to V_K and declined to a plateau of approximately half the maximal level at positive voltages. Voltage-dependent GIRK1 availability shifted with V_K and was half maximal at V_K -20 mV; the equivalent gating charge was ~1.6 e^-. The voltage-dependent gating parameters of GIRK1 did not significantly differ for G protein activation by three heterologously expressed signaling pathways: m2 muscarinic receptors, serotonin 1A receptors, or G protein β1y2 subunits. Voltage dependence was also unaffected by agonist concentration. These results indicate that the voltage-dependent gating properties of GIRK1 are not due to extrinsic factors such as agonist-receptor interactions and G protein-channel coupling, but instead are analogous to the intrinsic gating behaviors of other inwardly rectifying K^+ channels

Evidence that neuronal G-protein-gated inwardly rectifying K+ channels are activated by Gβγ subunits and function as heteromultimers

Guanine nucleotide-binding proteins (G proteins) activate K+ conductances in cardiac atrial cells to slow heart rate and in neurons to decrease excitability. cDNAs encoding three isoforms of a G-protein-coupled, inwardly rectifying K+ channel (GIRK) have recently been cloned from cardiac (GIRK1/Kir 3.1) and brain cDNA libraries (GIRK2/Kir 3.2 and GIRK3/Kir 3.3). Here we report that GIRK2 but not GIRK3 can be activated by G protein subunits Gβ1 and G2 in Xenopus oocytes. Furthermore, when either GIRK3 or GIRK2 was coexpressed with GIRK1 and activated either by muscarinic receptors or by Gβ subunits, G-protein-mediated inward currents were increased by 5- to 40-fold. The single-channel conductance for GIRK1 plus GIRK2 coexpression was intermediate between those for GIRK1 alone and for GIRK2 alone, and voltage-jump kinetics for the coexpressed channels displayed new kinetic properties. On the other hand, coexpression of GIRK3 with GIRK2 suppressed the GIRK2 alone response. These studies suggest that formation of heteromultimers involving the several GIRKs is an important mechanism for generating diversity in expression level and function of neurotransmitter-coupled, inward rectifier K+ channels

Studies of the structure of the plasmasphere as seen by radiosounder measurements aboard the Alovetti-satellite

The structure of the plasmasphere was studied as seen by radiosounder measurements aboard the Alovetti-2 satellite. Magnetic tape data files were obtained from the NASA Ames Research Center to give a reasonably complete set of high latitude electron density profiles. Considerable effort was expended to develop models of ion flow in the topside ionosphere. These models took both H(+) and O(+) into account and permitted various parameter studies to be made of the various factors which affect H(+) escape in polar wind flows. The results of these studies are included. Extensive computer programs were written to display the measured electron density profiles in ways useful to geophysical analysis. The expected mid-latitude trough was easily discernable in the nightime ionosphere at locations expected from similar observations of the plasmapause. In the dayside ionosphere, however, it proved extremely difficult to find any trough-like phenomena. Using the previously developed computer models, it was possible to study the region where the plasmapause appeared to be absent. It was found that over much of the dayside, large fluxes were computed well inside the plasmapause extending down to L-shells as low as 2.5

Behavior of thermal plasma in the ionosphere and magnetosphere

Models of ion flow in the topside ionosphere were developed. These models took both H(+) and O(+) into account and permitted various parameter studies to be made affecting H(+) escape in polar winds. Extensive computer programs were written to display the measured electron density profiles in ways useful to geophysical analysis. The relationship between the location of the plasmapause as it is found in the equatorial plane and the location of the ionospheric trough was also investigated

RGS4 regulates partial agonism of the M2 muscarinic receptor-activated K+ currents.

Partial agonists are used clinically to avoid overstimulation of receptor-mediated signalling, as they produce a submaximal response even at 100% receptor occupancy. The submaximal efficacy of partial agonists is due to conformational change of the agonist-receptor complex, which reduces effector activation. In addition to signalling activators, several regulators help control intracellular signal transductions. However, it remains unclear whether these signalling regulators contribute to partial agonism. Here we show that regulator of G-protein signalling (RGS) 4 is a determinant for partial agonism of the M2 muscarinic receptor (M2R). In rat atrial myocytes, pilocarpine evoked smaller G-protein-gated K(+) inwardly rectifying (KG) currents than those evoked by ACh. In a Xenopus oocyte expression system, pilocarpine acted as a partial agonist in the presence of RGS4 as it did in atrial myocytes, while it acted like a full agonist in the absence of RGS4. Functional couplings within the agonist-receptor complex/G-protein/RGS4 system controlled the efficacy of pilocarpine relative to ACh. The pilocarpine-M2R complex suppressed G-protein-mediated activation of KG currents via RGS4. Our results demonstrate that partial agonism of M2R is regulated by the RGS4-mediated inhibition of G-protein signalling. This finding helps us to understand the molecular components and mechanism underlying the partial agonism of M2R-mediated physiological responses

Evidence for a Functional Interaction between Integrins and G Protein-activated Inward Rectifier K+ Channels

Heteromultimeric G protein-activated inward rectifier K+ (GIRK) channels, abundant in heart and brain, help to determine the cellular membrane potential as well as the frequency and duration of electrical impulses. The sequence arginine-glycine-aspartate (RGD), located extracellularly between the first membrane-spanning region and the pore, is conserved among all identified GIRK subunits but is not found in the extracellular domain of any other cloned K+ channels. Many integrins, which, like channels, are integral membrane proteins, recognize this RGD sequence on other proteins, usually in the extracellular matrix. We therefore asked whether GIRK activity might be regulated by direct interaction with integrin. Here, we present evidence that mutation of the RGD site to RGE, particularly on the GIRK4 subunit, decreases or abolishes GIRK current. Furthermore, wild-type channels can be co-immunoprecipitated with integrin. The total cellular amount of expressed mutant GIRK channel protein is the same as the wild-type protein; however, the amount of mutant channel protein that localizes to the plasma membrane is decreased relative to wild-type, most likely accounting for the diminished GIRK current detected. GIRK channels appear to bind directly to integrin and to require this interaction for proper GIRK channel membrane localization and function

Proceedings of the 1st Space Plasma Computer Analysis Network (SCAN) Workshop

The purpose of the workshop was to identify specific cooperative scientific study topics within the discipline of Ionosphere Magnetosphere Coupling processes and to develop methods and procedures to accomplish this cooperative research using SCAN facilities. Cooperative scientific research was initiated in the areas of polar cusp composition, O+ polar outflow, and magnetospheric boundary morphology studies and an approach using a common metafile structure was adopted to facilitate the exchange of data and plots between the various workshop participants. The advantages of in person versus remote workshops were discussed also

Probing the role of the cation–π interaction in the binding sites of GPCRs using unnatural amino acids

We describe a general application of the nonsense suppression methodology for unnatural amino acid incorporation to probe drug–receptor interactions in functional G protein-coupled receptors (GPCRs), evaluating the binding sites of both the M2 muscarinic acetylcholine receptor and the D2 dopamine receptor. Receptors were expressed in Xenopus oocytes, and activation of a G protein-coupled, inward-rectifying K^+ channel (GIRK) provided, after optimization of conditions, a quantitative readout of receptor function. A number of aromatic amino acids thought to be near the agonist-binding site were evaluated. Incorporation of a series of fluorinated tryptophan derivatives at W6.48 of the D2 receptor establishes a cation–π interaction between the agonist dopamine and W6.48, suggesting a reorientation of W6.48 on agonist binding, consistent with proposed “rotamer switch” models. Interestingly, no comparable cation–π interaction was found at the aligning residue in the M2 receptor

Preliminary studies of pest constraints to cotton seedlings in a direct seeding mulch-based system in Cameroon

The present study evaluated the pest constraints of an innovative crop management system in Cameroon involving conservation tillage and direct seeding mulch-based strategies. We hypothesized that the presence of mulch (i) would support a higher density of phytophagous arthropods particularly millipedes as well as pathogenic fungi that cause severe damage to cotton seedlings and (ii) would reduce early aphid infestations. The impact of two cover-crop mulches Calopogonium mucunoides and Brachiaria ruziziensis on the vigour of seedling cotton stands and arthropod damage was assessed in two independent field experiments conducted in 2001 and 2002 respectively. In both experiments the presence of mulch negatively affected cotton seedling stand by 13–14% compared to non-mulched plots and the proportion of damaged seedlings was higher in mulched than in non-mulched plots supporting the hypothesis that mulch favoured soil pest damage. In both experiments insecticidal seed dressing increased the seedling stand and the number of dead millipedes collected and fungicide had little or no effect on seedling stand and vigour. It was however observed in 2002 that the fungicide seed dressing had a positive effect on seedling stand in non-mulched plots but not in mulched plots suggesting that fungi may have been naturally inhibited by B. ruziziensis mulch. The dynamics of aphid colonization was not influenced by the presence of mulch. In 2001 taller seedlings were found in mulched than non-mulched plots probably due to greater water and nutrient availability in C. mucunoides-mulched plots than in non-mulched plot

Information sectorielle : quelle place dans les rapports des analystes financiers ? Une étude empirique du secteur hôtelier international

Notre recherche propose d'identifier comment les analystes se réfèrent à l'information sectorielle publiée par les groupes et quelle place occupe l'information sectorielle dans leurs modèles financiers. Nombreuses sont les recherches qui se fondent sur l'analyse des consensus, peu s'intéressent au contenu réel des rapports réalisés par les analystes. Nous avons déterminé les références faites à l'information sectorielle tant obligatoire que volontaire au sein des rapports rédigés en 2006 par les analystes financiers des groupes hôteliers internationaux publiant selon la norme SFAS 131 et IAS 14. Nous avons aussi observé si les rapports des analystes financiers présentaient des modèles de prévisions ou de valorisations construits selon une base sectorielle. Nos résultats montrent que les analystes financiers se réfèrent très massivement à l'information sectorielle, qu'il s'agisse de l'information normalisée ou d'une information volontaire. Plus spécifiquement, si elle est disponible, l'information géographique sectorielle est très souvent utilisée. Toutefois, l'approche sectorielle demeure occasionnelle dans les modèles utilisés pour argumenter les prévisions ou les valorisations ; l'approche selon les « secteurs d'activité » étant alors privilégiée. La pertinence des résultats obtenus d'après une approche sectorielle dans les modèles financiers pourra faire l'objet d'une recherche future. La récente adoption de la norme IFRS 8, convergence des référentiels IFRS et US GAAP, pourrait bien aussi impacter les outils de modélisation utilisés par les analystes.Information sectorielle, analystes financiers, groupes hôteliers