Comparison of Sex Pheromone and Kairomone-Enhanced Pheromone Lures for Monitoring Oriental Fruit Moth (Lepidoptera: Tortricidae) in Mating Disruption and Non-Disruption Tree Fruit Orchards.

Oriental fruit moth, Grapholita molesta (Busck), populations were monitored using standard sex pheromone lures (OFM L2) and kairomone-enhanced lures to aid the interpretation of trap captures with enhanced relative to conventional lures. Initially, comparison of 10 different lures showed that a10X load of OFM pheromone, codlemone, terpinyl acetate, and acetic acid were key components of the most attractive lures (TRE11034 and 1123). Subsequent trapping studies in mating disruption and non-disrupted orchards in the United States and Spain compared trap captures with TRE1123 and OFM L2 lures. Compared to the OFM L2 lure, the TRE1123 lure captured more moths in mating disruption and non-disrupted orchards, caught female moths, improved the precision of mean population estimates, and led to greater resolution of generational flights. Suppression of trap captures in mating disruption versus non-disrupted orchards was similar with both lures. There were significant linear correlations between weekly trap captures with the two lures in the majority of mating disruption and non-disrupted orchards across locations and years. Furthermore, regression of the slopes of trap capture regressions (i.e., attractiveness of enhanced lures relative to sex pheromone lures alone) versus moth density (as measured by mean cumulative moth capture with TRE1123 and OFM L2 lures) exhibited a significant positive relationship in non-disrupted orchards, indicating enhanced lures were relatively more attractive under high population densities. This relationship was not significant in mating disruption orchards, likely due to the density independent, non-competitive mechanism of mating disruption for oriental fruit moth when using high-dose reservoir dispensers.

Functional recruitment of dynamin requires multimeric interactions for efficient endocytosis.

During clathrin mediated endocytosis (CME), the concerted action of dynamin and its interacting partners drives membrane scission. Essential interactions occur between the proline/arginine-rich domain of dynamin (dynPRD) and the Src-homology domain 3 (SH3) of various proteins including amphiphysins. Here we show that multiple SH3 domains must bind simultaneously to dynPRD through three adjacent motifs for dynamin's efficient recruitment and function. First, we show that mutant dynamins modified in a single motif, including the central amphiphysin SH3 (amphSH3) binding motif, partially rescue CME in dynamin triple knock-out cells. However, mutating two motifs largely prevents that ability. Furthermore, we designed divalent dynPRD-derived peptides. These ligands bind multimers of amphSH3 with >100-fold higher affinity than monovalent ones in vitro. Accordingly, dialyzing living cells with these divalent peptides through a patch-clamp pipette blocks CME much more effectively than with monovalent ones. We conclude that dynamin drives vesicle scission via multivalent interactions in cells.

Engineering selective competitors for the discrimination of highly conserved protein-protein interaction modules.

Designing highly specific modulators of protein-protein interactions (PPIs) is especially challenging in the context of multiple paralogs and conserved interaction surfaces. In this case, direct generation of selective and competitive inhibitors is hindered by high similarity within the evolutionary-related protein interfaces. We report here a strategy that uses a semi-rational approach to separate the modulator design into two functional parts. We first achieve specificity toward a region outside of the interface by using phage display selection coupled with molecular and cellular validation. Highly selective competition is then generated by appending the more degenerate interaction peptide to contact the target interface. We apply this approach to specifically bind a single PDZ domain within the postsynaptic protein PSD-95 over highly similar PDZ domains in PSD-93, SAP-97 and SAP-102. Our work provides a paralog-selective and domain specific inhibitor of PSD-95, and describes a method to efficiently target other conserved PPI modules.

Modulation of AMPA receptor surface diffusion restores hippocampal plasticity and memory in Huntington's disease models.

Impaired hippocampal synaptic plasticity contributes to cognitive impairment in Huntington's disease (HD). However, the molecular basis of such synaptic plasticity defects is not fully understood. Combining live-cell nanoparticle tracking and super-resolution imaging, we show that AMPAR surface diffusion, a key player in synaptic plasticity, is disturbed in various rodent models of HD. We demonstrate that defects in the brain-derived neurotrophic factor (BDNF)-tyrosine receptor kinase B (TrkB) signaling pathway contribute to the deregulated AMPAR trafficking by reducing the interaction between transmembrane AMPA receptor regulatory proteins (TARPs) and the PDZ-domain scaffold protein PSD95. The disturbed AMPAR surface diffusion is rescued by the antidepressant drug tianeptine via the BDNF signaling pathway. Tianeptine also restores the impaired LTP and hippocampus-dependent memory in different HD mouse models. These findings unravel a mechanism underlying hippocampal synaptic and memory dysfunction in HD, and highlight AMPAR surface diffusion as a promising therapeutic target.

Differential Nanoscale Topography and Functional Role of GluN2-NMDA Receptor Subtypes at Glutamatergic Synapses.

NMDA receptors (NMDARs) play key roles in the use-dependent adaptation of glutamatergic synapses underpinning memory formation. In the forebrain, these plastic processes involve the varied contributions of GluN2A- and GluN2B-containing NMDARs that have different signaling properties. Although the molecular machinery of synaptic NMDAR trafficking has been under scrutiny, the postsynaptic spatial organization of these two receptor subtypes has remained elusive. Here, we used super-resolution imaging of NMDARs in rat hippocampal synapses to unveil the nanoscale topography of native GluN2A- and GluN2B-NMDARs. Both subtypes were found to be organized in separate nanodomains that vary over the course of development. Furthermore, GluN2A- and GluN2B-NMDAR nanoscale organizations relied on distinct regulatory mechanisms. Strikingly, the selective rearrangement of GluN2A- and GluN2B-NMDARs, with no overall change in NMDAR current amplitude, allowed bi-directional tuning of synaptic LTP. Thus, GluN2A- and GluN2B-NMDAR nanoscale organizations are differentially regulated and seem to involve distinct signaling complexes during synaptic adaptation.

Pre-post synaptic alignment through neuroligin-1 tunes synaptic transmission efficiency.

The nanoscale organization of neurotransmitter receptors regarding pre-synaptic release sites is a fundamental determinant of the synaptic transmission amplitude and reliability. How modifications in the pre- and post-synaptic machinery alignments affects synaptic currents, has only been addressed with computer modelling. Using single molecule super-resolution microscopy, we found a strong spatial correlation between AMPA receptor (AMPAR) nanodomains and the post-synaptic adhesion protein neuroligin-1 (NLG1). Expression of a truncated form of NLG1 disrupted this correlation without affecting the intrinsic AMPAR organization, shifting the pre-synaptic release machinery away from AMPAR nanodomains. Electrophysiology in dissociated and organotypic hippocampal rodent cultures shows these treatments significantly decrease AMPAR-mediated miniature and EPSC amplitudes. Computer modelling predicts that ~100 nm lateral shift between AMPAR nanoclusters and glutamate release sites induces a significant reduction in AMPAR-mediated currents. Thus, our results suggest the synapses necessity to release glutamate precisely in front of AMPAR nanodomains, to maintain a high synaptic responses efficiency.

CaMKII Metaplasticity Drives Aß Oligomer-Mediated Synaptotoxicity.

Alzheimer's disease (AD) is emerging as a synaptopathology driven by metaplasticity. Indeed, reminiscent of metaplasticity, oligomeric forms of the amyloid-ß peptide (oAß) prevent induction of long-term potentiation (LTP) via the prior activation of GluN2B-containing NMDA receptors (NMDARs). However, the downstream Ca2+-dependent signaling molecules that mediate aberrant metaplasticity are unknown. In this study, we show that oAß promotes the activation of Ca2+/calmodulin-dependent kinase II (CaMKII) via GluN2B-containing NMDARs. Importantly, we find that CaMKII inhibition rescues both the LTP impairment and the dendritic spine loss mediated by oAß. Mechanistically resembling metaplasticity, oAß prevents subsequent rounds of plasticity from inducing CaMKII T286 autophosphorylation, as well as the associated anchoring and accumulation of synaptic AMPA receptors (AMPARs). Finally, prolonged oAß treatment-induced CaMKII misactivation leads to dendritic spine loss via the destabilization of surface AMPARs. Thus, our study demonstrates that oAß engages synaptic metaplasticity via aberrant CaMKII activation.

Determination of the Baseline Susceptibility of European Populations of Cydia pomonella (Lepidoptera: Tortricidae) to Chlorantraniliprole and the Role of Cytochrome P450 Monooxygenases.

The codling moth, Cydia pomonella (L.) (Lepidoptera: Tortricidae), is the key pest on pome fruit and walnut orchards worldwide. Its resistance to available insecticides has been widely reported. Chlorantraniliprole is an anthranilic diamide that was introduced in European countries in 2008-2009 and acts by activating the insect's ryanodine receptors. The aims of this study were to determine the baseline susceptibility of European populations of C. pomonella to chlorantraniliprole, to establish the discriminant concentrations (DC) to check the possible development of resistance, and to know the role of cytochrome P450 monooxygenases (P450) in the possible susceptibility decrease of field populations to the insecticide. Ten field populations from Spain along with others were used to calculate the baseline response of larvae to chlorantraniliprole incorporated into the diet. A pooled probit line was calculated, and three DC were established: 0.3 mg a.i./kg (close to the LC50), 1.0 mg a.i./kg (close to the LC90), and 10 mg a.i./kg diets (threefold the LC99). The DC were used to test the susceptibility of 27 field populations from France, Germany, Hungary, Italy, and Spain. The corrected mortality observed in all cases ranged within the expected interval, even with Spanish populations that showed between 12.1 and 100.0% of individuals with high P450 activity. However, the mortality caused by the DC0.3 decreased as the mean P450 activity increased. Field populations resistant to other insecticides were susceptible to chlorantraniliprole. The determined baseline codling moth susceptibility is a valuable reference for tracking possible future alterations in the efficacy of the insecticide.

Monitoring resistance of Cydia pomonella (L.) Spanish field populations to new chemical insecticides and the mechanisms involved.

BACKGROUND: Widespread resistance of Cydia pomonella to organophosphates was demonstrated in populations from the Spanish Ebro Valley area which showed high levels of enzymatic detoxification. To determine the efficacy of new insecticides, neonate larval bioassays were carried out on 20 field codling moth populations collected from three different Spanish apple production areas. Synergist bioassays were performed to determine the enzymatic mechanisms involved. RESULTS: The least active ingredients were methoxyfenozide, with 100% of the populations showing significantly lower mortality than the susceptible strain, and lambda-cyhalothrin, with very high resistance ratios (872.0 for the most resistant field population). Approximately 50% of the populations were resistant or tolerant to thiacloprid. By contrast, tebufenozide was very effective in all the field populations, as was chlorpyrifos-ethyl despite its widespread use during the last few years. Indoxacarb, spinosad and chlorantraniliprole also provided high efficacy, as did emamectin and spinetoram, which are not yet registered in Spain. CONCLUSION: The resistant Spanish codling moth populations can be controlled using new reduced-risk insecticides. The use of synergists showed the importance of the concentration applied and the difficulty of interpreting results in field populations that show multiple resistance to different active ingredients. © 2017 Society of Chemical Industry.

Comparative Effect of Three Neurotoxic Insecticides With Different Modes of Action on Adult Males and Females of Three Tortricid Moth Pests.

Insecticides are the dominant pest management method in fruit and vegetable crops worldwide owing to their quick effect, low cost, and relatively easy application, but they bear negative effects on human health and the environment. Insecticide mode of action (MoA), target species, and sex are variables that could affect insecticide-induced mortality. We recorded the mortality caused by three neurotoxic insecticides with different modes of action (chlorpyrifos [organophosphate, acetylcholinesterase inhibitor], λ-cyhalothrin [pyrethroid, sodium channel modulator], and thiacloprid [neonicotinoid, nicotinic acetylcholinesterase receptor agonist]) applied topically to adult males and females of three economically important tortricid species [Cydia pomonella (L.), Grapholita molesta (Busck), and Lobesia botrana (Denis & Schiffermüller)] that strongly depend on insecticide use for their control. Concentration and dose-mortality curves were recorded at 24 and 48 h postapplication. Large mortality differences between insecticides (maximum 7,800-fold for LD50) were followed by much lower, yet important, differences between species (maximum 115-fold), and sexes (maximum 41.5-fold). Significant interactions between the three factors indicate that they are not independent from each other. Interestingly, with the organophosphate chlorpyrifos, males of the three species were less susceptible than females, which was unexpected, as females are larger than males. Higher female sensitivity to organophosphates has been reported previously but only in G. molesta, not in other moth species. Our results highlight the importance of taking into account sex in dose-mortality studies with adult moths.

Lengthening of the Stargazin Cytoplasmic Tail Increases Synaptic Transmission by Promoting Interaction to Deeper Domains of PSD-95.

PSD-95 is a prominent organizer of the postsynaptic density (PSD) that can present a filamentous orientation perpendicular to the plasma membrane. Interactions between PSD-95 and transmembrane proteins might be particularly sensitive to this orientation, as "long" cytoplasmic tails might be required to reach deeper PSD-95 domains. Extension/retraction of transmembrane protein C-tails offer a new way of regulating binding to PSD-95. Using stargazin as a model, we found that enhancing the apparent length of stargazin C-tail through phosphorylation or by an artificial linker was sufficient to potentiate binding to PSD-95, AMPAR anchoring, and synaptic transmission. A linear extension of stargazin C-tail facilitates binding to PSD-95 by preferentially engaging interaction with the farthest located PDZ domains regarding to the plasma membrane, which present a greater affinity for the stargazin PDZ-domain-binding motif. Our study reveals that the concerted orientation of the stargazin C-tail and PSD-95 is a major determinant of synaptic strength.

Intraspecific Variation in Female Sex Pheromone of the Codling Moth Cydia pomonella.

The codling moth, Cydia pomonella L. (Lepidoptera, Tortricidae), is a major pest of apple, pear and walnut orchards worldwide. This pest is often controlled using the biologically friendly control method known as pheromone-based mating disruption. Mating disruption likely exerts selection on the sexual communication system of codling moth, as male and female moths will persist in their attempt to meet and mate. Surprisingly little is known on the intraspecific variation of sexual communication in this species. We started an investigation to determine the level of individual variation in the female sex pheromone composition of this moth and whether variation among different populations might be correlated with use of mating disruption against those populations. By extracting pheromone glands of individual females from a laboratory population in Canada and from populations from apple orchards in Spain and Italy, we found significant between- and within-population variation. Comparing females that had been exposed to mating disruption, or not, revealed a significant difference in sex pheromone composition for two of the minor components. Overall, the intraspecific variation observed shows the potential for a shift in female sexual signal when selection pressure is high, as is the case with continuous use of mating disruption.

Neurexin-1ß binding to neuroligin-1 triggers the preferential recruitment of PSD-95 versus gephyrin through tyrosine phosphorylation of neuroligin-1.

Adhesion between neurexin-1ß (Nrx1ß) and neuroligin-1 (Nlg1) induces early recruitment of the postsynaptic density protein 95 (PSD-95) scaffold; however, the associated signaling mechanisms are unknown. To dissociate the effects of ligand binding and receptor multimerization, we compared conditions in which Nlg1 in neurons was bound to Nrx1ß or nonactivating HA antibodies. Time-lapse imaging, fluorescence recovery after photobleaching, and single-particle tracking demonstrated that in addition to aggregating Nlg1, Nrx1ß binding stimulates the interaction between Nlg1 and PSD-95. Phosphotyrosine immunoblots and pull-down of gephyrin by Nlg1 peptides in vitro showed that Nlg1 can be phosphorylated at a unique tyrosine (Y782), preventing gephyrin binding. Expression of Nlg1 point mutants in neurons indicated that Y782 phosphorylation controls the preferential binding of Nlg1 to PSD-95 versus gephyrin, and accordingly the formation of inhibitory and excitatory synapses. We propose that ligand-induced changes in the Nlg1 phosphotyrosine level control the balance between excitatory and inhibitory scaffold assembly during synapse formation and stabilization.

Electrophilic derivatives antagonise pheromone attraction in Cydia pomonella.

BACKGROUND: Pheromone antagonists are good disruptants of the pheromone communication in insects and, as such, have been used in mating disruption experiments. In this study, new non-fluorinated electrophilic keto derivatives structurally related to the pheromone of Cydia pomonella (codlemone) have been synthesised and tested as putative pheromone antagonists. RESULTS: Codlemone (1) was prepared in excellent stereoselectivity in a new, iterative approach involving two Horner-Wadsworth-Emmons reactions. Methyl ketone (2), keto ester (3) and diketone (4) were obtained from codlemone in straightforward approaches in good overall yields and excellent stereochemical purity (≥98% E,E). In electrophysiology, only compound 2 displayed inhibition of the antennal response to the pheromone after presaturation of the antennal receptors. Compounds 2 to 4 did not inhibit the pheromone-degrading enzyme responsible for codlemone metabolism, but mixtures of ketone 2 and diketone 4 with codlemone elicited erratic flights on males in a wind tunnel. In the field, blends of either compound (2 or 4) with the pheromone caught significantly fewer males than codlemone alone. CONCLUSION: Codlemone and the potential antagonists 2 to 4 have been synthesised in good yields and excellent stereoselectivity. These chemicals behave as pheromone antagonists of the codling moth both in the laboratory and in the field.

Inhibition of PDZ domain-mediated interactions.

Modulating protein-protein interactions constitutes a promising strategy both for the investigation of biological mechanisms and for developing new therapeutic approaches. Among the many types of inter-actions, PDZ domain-mediated interactions (PDMIs) have emerged over the last decade as attractive targets in the drug discovery field. Indeed, these small domains are involved in the regulation of many signaling pathways and possess structural properties which are favorable for the design of competing ligands. Herein, we describe the recent approaches developed to inhibit this class of protein-protein interactions.

Applications of 3-aminolactams: design, synthesis, and biological evaluation of a library of potential dimerisation inhibitors of HIV1-protease.

In the context of our studies on the applications of 3-aminolactams as conformationally restricted pseudodipeptides, we report here the synthesis of a library of potential dimerisation inhibitors of HIV1-protease. Two of the pseudopeptides were active on the wild type virus (HIV1) at micromolar levels (EC(50)). Although the peptides showed lower anti-viral activity than previously reported dimerisation inhibitors, our results demonstrate that the piperidone moiety does not prevent cell penetration, and hence that such derivatization is compatible with potential anti-HIV treatment.

Synthesis of allylic trifluoromethyl ketones and their activity as inhibitors of the sex pheromone of the leopard moth, Zeuzera pyrina L. (Lepidoptera: Cossidae).

BACKGROUND: Trifluoromethyl ketones (TFMKs), structurally related to the pheromones, are good inhibitors of pheromone communication in insects. To determine their activity on Zeuzera pyrina L. (Lepidoptera: Cossidae), a polyphagous pest, the authors have prepared two diunsaturated TFMK analogues of the major (3) and the minor (4) pheromone components, and two monounsaturated ones (5, 6). Their biological activity in electroantennogram (EAG), wind tunnel and field tests is presented. RESULTS: The synthetic strategy to obtain the allylic TFMKs 3 and 5 is based on the reactions of diene 10 and 1-octadecene with trifluoroacetaldehyde ethyl hemiacetal, followed by Dess-Martin oxidation of the resulting homoallylic trifluoromethyl alcohols. In EAG, topical application of analogues 3 and 4 on male antennae significantly reduced the pheromone response. In the wind tunnel, compound 4 reduced the number of contacts with the pheromone source. In the field, traps baited with mixtures of pheromone and inhibitors captured significantly fewer males than the pheromone alone. CONCLUSION: An efficient synthesis of allylic TFMKs is reported, with good overall yield, regiospecificity and diastereoselectivity. These compounds are good inhibitors of the pheromone in electrophysiology, wind tunnel and field tests. The results show the importance of two unsaturations at positions 2 and 13 of the trifluoroacyl group in the structure of the analogues, the latter being critical for inhibitory activity.

Improved Fmoc-based solid-phase synthesis of homologous peptide fragments of human and mouse prion proteins.

The synthesis of difficult peptide sequences has been a challenge since the very beginning of SPPS. The self-assembly of the growing peptide chains has been proposed as one of the causes of this synthetic problem. However, there is an increasing need to obtain peptides and proteins that are prone to aggregate. These peptides and proteins are generally associated with diseases known as amyloidoses. We present an efficient SPPS of two homologous peptide fragments of HuPrP (106-126) and MoPrP105-125 based on the use of the PEGA resin combined with proper coupling approaches. These peptide fragments were also studied by CD and TEM to determine their ability to aggregate. On the basis of these results, we support PEG-based resins as an efficient synthetic tool to prepare peptide sequences prone to aggregate on-resin.