379 research outputs found

    Agonist and antagonist properties of an insect GABA-gated chloride channel (RDL) are influenced by heterologous expression conditions

    Get PDF
    Pentameric ligand-gated ion channels (pLGICs) activated by the inhibitory neurotransmitter γ-aminobutyric acid (GABA) are expressed widely in both vertebrate and invertebrate species. One of the best characterised insect GABA-gated chloride channels is RDL, an abbreviation of 'resistance to dieldrin', that was originally identified by genetic screening in Drosophila melanogaster. Here we have cloned the analogous gene from the bumblebee Bombus terrestris audax (BtRDL) and examined its pharmacological properties by functional expression in Xenopus oocytes. Somewhat unexpectedly, the sensitivity of BtRDL to GABA, as measured by its apparent affinity (EC50), was influenced by heterologous expression conditions. This phenomenon was observed in response to alterations in the amount of cRNA injected; the length of time that oocytes were incubated before functional analysis; and by the presence or absence of a 3' untranslated region. In contrast, similar changes in expression conditions were not associated with changes in apparent affinity with RDL cloned from D. melanogaster (DmRDL). Changes in apparent affinity with BtRDL were also observed following co-expression of a chaperone protein (NACHO). Similar changes in apparent affinity were observed with an allosteric agonist (propofol) and a non-competitive antagonist (picrotoxinin), indicating that expression-depended changes are not restricted to the orthosteric agonist binding site. Interestingly, instances of expression-dependent changes in apparent affinity have been reported previously for vertebrate glycine receptors, which are also members of the pLGIC super-family. Our observations with BtRDL are consistent with previous data obtained with vertebrate glycine receptors and indicates that agonist and antagonist apparent affinity can be influenced by the level of functional expression in a variety of pLGICs

    Recurrent Gaussian processes

    Get PDF
    We define Recurrent Gaussian Processes (RGP) models, a general family of Bayesian nonparametric models with recurrent GP priors which are able to learn dynamical patterns from sequential data. Similar to Recurrent Neural Networks (RNNs), RGPs can have different formulations for their internal states, distinct inference methods and be extended with deep structures. In such context, we propose a novel deep RGP model whose autoregressive states are latent, thereby performing representation and dynamical learning simultaneously. To fully exploit the Bayesian nature of the RGP model we develop the Recurrent Variational Bayes (REVARB) framework, which enables efficient inference and strong regularization through coherent propagation of uncertainty across the RGP layers and states. We also introduce a RGP extension where variational parameters are greatly reduced by being reparametrized through RNN-based sequential recognition models. We apply our model to the tasks of nonlinear system identification and human motion modeling. The promising obtained results indicate that our RGP model maintains its highly flexibility while being able to avoid overfitting and being applicable even when larger datasets are not available

    Mutations on M3 helix of Plutella xylostella glutamate-gated chloride channel confer unequal resistance to abamectin by two different mechanisms

    Get PDF
    Abamectin is one of the most widely used avermectins for agricultural pests control, but the emergence of resistance around the world is proving a major threat to its sustained application. Abamectin acts by directly activating glutamate-gated chloride channels (GluCls) and modulating other Cys-loop ion channels. To date, three mutations occurring in the transmembrane domain of arthropod GluCls are associated with target-site resistance to abamectin: A309V in Plutella xylostella GluCl (PxGluCl), G323D in Tetranychus urticae GluCl1 (TuGluCl1) and G326E in TuGluCl3. To compare the effects of these mutations in a single system, A309V/I/G and G315E (corresponding to G323 in TuGluCl1 and G326 in TuGluCl3) substitutions were introduced individually into the PxGluCl channel. Functional analysis using Xenopus oocytes showed that the A309V and G315E mutations reduced the sensitivity to abamectin by 4.8- and 493-fold, respectively. In contrast, the substitutions A309I/G show no significant effects on the response to abamectin. Interestingly, the A309I substitution increased the channel sensitivity to glutamate by one order of magnitude (∼12-fold). Analysis of PxGluCl homology models indicates that the G315E mutation interferes with abamectin binding through a steric hindrance mechanism. In contrast, the structural consequences of the A309 mutations are not so clear and an allosteric modification of the binding site is the most likely mechanism. Overall the results show that both A309V and G315E mutations may contribute to target-site resistance to abamectin and may be important for the future prediction and monitoring of abamectin resistance in P. xylostella and other arthropod pests

    Evaluation of the postharvest quality of Cagaita fruits (Eugenia dysenterica DC.) coated with chitosan and associated with refrigeration

    Get PDF
    Cagaita fruits are subject to seasonality and perishability. This work aims to use scanning electron microscopy (SEM) to evaluate the physicochemical characteristics, texture, color and physical structure of cagaita fruits coated with different chitosan concentrations. The fruits were divided as follows: T0 (uncoated fruits), T1 (fruits coated with 1% (v/v) chitosan), T2 (fruits coated with 2% (v/v) chitosan) and T3 (fruits coated with 3% (v/v) chitosan). They were analyzed at 0, 10, 20 and 30 days of storage. Titratable acidity and soluble solids content showed no conservation of fruit characteristics; they showed better results for uncoated fruits, as well as weight loss, vitamin C and peak strain. The color of cagaita fruits confirmed ripening during storage regardless of treatment. Scanning electron microscopy showed that the film solution did not adhere, as desired, to the cell wall of fruits. As the results of fruits coated with 3% pectin were close to control, further studies should be carried out with higher coating percentages so that the fruit quality is maintained during storage.Keywords: Physical structure, film solution, quality, shelf life

    Characterizing photonic crystal waveguides with an expanded k-space evanescent coupling technique

    Full text link
    We demonstrate a direct, single measurement technique for characterizing the dispersion of a photonic crystal waveguide (PCWG) using a tapered fiber evanescent coupling method. A highly curved fiber taper is used to probe the Fabry-Pérot spectrum of a closed PCWG over a broad k-space range, and from this measurement the dispersive properties of the waveguide can be found. Waveguide propagation losses can also be estimated from measurements of closed waveguides with different lengths. The validity of this method is demonstrated by comparing the results obtained on a 'W1' PCWG in chalcogenide glass with numerical simulation. © 2008 Optical Society of America
    • …
    corecore