Carboxyl-terminal Truncations of ClC-Kb Abolish Channel Activation by Barttin Via Modified Common Gating and Trafficking

ClC-K chloride channels are crucial for auditory transduction and urine concentration. Mutations in CLCNKB, the gene encoding the renal chloride channel hClC-Kb, cause Bartter syndrome type III, a human genetic condition characterized by polyuria, hypokalemia, and alkalosis. In recent years, several Bartter syndrome-associated mutations have been described that result in truncations of the intracellular carboxyl terminus of hClC-Kb. We here used a combination of whole-cell patch clamp, confocal imaging, co-immunoprecipitation, and surface biotinylation to study the functional consequences of a frequent CLCNKB mutation that creates a premature stop codon at Trp-610. We found that W610X leaves the association of hClC-Kb and the accessory subunit barttin unaffected, but impairs its regulation by barttin. W610X attenuates hClC-Kb surface membrane insertion. Moreover, W610X results in hClC-Kb channel opening in the absence of barttin and prevents further barttin-mediated activation. To describe how the carboxyl terminus modifies the regulation by barttin we used V166E rClC-K1. V166E rClC-K1 is active without barttin and exhibits prominent, barttin-regulated voltage-dependent gating. Electrophysiological characterization of truncated V166E rClC-K1 demonstrated that the distal carboxyl terminus is necessary for slow cooperative gating. Since barttin modifies this particular gating process, channels lacking the distal carboxyl-terminal domain are no longer regulated by the accessory subunit. Our results demonstrate that the carboxyl terminus of hClC-Kb is not part of the binding site for barttin, but functionally modifies the interplay with barttin. The loss-of-activation of truncated hClC-Kb channels in heterologous expression systems fully explains the reduced basolateral chloride conductance in affected kidneys and the clinical symptoms of Bartter syndrome patients.     

Individually mark–mass release–resight study elucidates effects of patch characteristics and distance on host patch location by an insect herbivore

1. How organisms locate their hosts is of fundamental importance in a variety of basic and applied ecological fields, including population dynamics, invasive species management and biological control. However, tracking movement of small organisms, such as insects, poses significant logistical challenges. 2. Mass‐release and individual–mark–recapture techniques were combined in an individually mark–mass release–resight (IMMRR) approach to track the movement of over 2000 adult insects in an economically important plant–herbivore system. Despite its widespread use for the biological control of the invasive thistle Carduus nutans, the host‐finding behaviour of the thistle head weevil Rhinocyllus conicus has not previously been studied. Insects were released at different distances from a mosaic of artificially created host patches with different areas and number of plants to assess the ecological determinants of patch finding. 3. The study was able to characterize the within‐season dispersal abilities and between‐patch movement patterns of R. conicus. Weevils found host plant patches over 900 m away. Large patches, with tall plants, situated close to the nearest release point had the highest first R. conicus resights. Patch area and plant density had no effect on the number of weevils resighted per plant; however, R. conicus individuals were more likely to disperse out of small patches and into large patches. 4. By understanding how R. conicus locates host patches of C. nutans, management activities for the control of this invasive thistle can be better informed. A deeper mechanistic understanding of host location will also improve prediction of coupled plant–herbivore spatial dynamics in general.     

基于无人机影像的阿拉善东南部荒漠灌丛植被空间格局研究

干旱区灌丛植被空间格局受多种物理和生态过程影响，能够指示生态系统的状态。研究通过量化灌丛斑块大小的空间分布来评估阿拉善高原东南部覆沙荒漠植被生态系统的状态，采用点格局分析法分析灌木种群的相互关系，以阐明不同灌木种在斑块格局形成中的作用，并结合土壤条件及下垫面粗糙度等指标验证评估的准确性，探讨灌丛空间格局差异的内在机理。结果表明，研究区样方2灌丛斑块大小符合截尾幂律分布，其他样方符合对数正态分布，前者的空间结构及生境条件均优于后者，说明植被空间格局可以准确表征生态系统状态。在局地尺度上灌木种内和种间呈现不同的相互关系，以竞争关系为主导是导致斑块破碎化的主要驱动机制。小灌木（如猫头刺）的种内互利关系有利于促进多样化斑块形态的形成，而大灌木（如沙冬青和蒙古扁桃）种间的互利作用则有利于形成异质性更强的复杂空间格局。基于灌丛斑块的空间格局评估生态系统状态，可为保护和恢复生态脆弱区受损植被提供重要的借鉴。     

Electrostatic networks control plug stabilization in the PapC usher

Abstract

The PapC usher, a β-barrel pore in the outer membrane of uropathogenic Escherichia coli, is used for assembly of the P pilus, a key virulence factor in bacterial colonization of human kidney cells. Each PapC protein is composed of a 24-stranded β-barrel channel, flanked by N- and C-terminal globular domains protruding into the periplasm, and occluded by a plug domain (PD). The PD is displaced from the channel towards the periplasm during pilus biogenesis, but the molecular mechanism for PD displacement remains unclear. Two structural features within the β-barrel, an α-helix and β5-6 hairpin loop, may play roles in controlling plug stabilization. Here we have tested clusters of residues at the interface of the plug, barrel, α-helix and hairpin, which participate in electrostatic networks. To assess the roles of these residues in plug stabilization, we used patch-clamp electrophysiology to compare the activity of wild-type and mutant PapC channels containing alanine substitutions at these sites. Mutations interrupting each of two salt bridge networks were relatively ineffective in disrupting plug stabilization. However, mutation of two pairs of arginines located at the inner and the outer surfaces of the PD resulted in an enhanced propensity for plug displacement. One arginine pair involved in a repulsive interaction between the linkers that tether the plug to the β-barrel was particularly sensitive to mutation. These results suggest that plug displacement, which is necessary for pilus assembly and translocation, may require a weakening of key electrostatic interactions between the plug linkers, and the plug and the α-helix.     

Energetics of gating MscS by membrane tension in azolectin liposomes and giant spheroplasts

Mechanosensitive (MS) ion channels are molecular sensors that detect and transduce signals across prokaryotic and eukaryotic cell membranes arising from external mechanical stimuli or osmotic gradients. They play an integral role in mechanosensory responses including touch, hearing, and proprioception by opening or closing in order to facilitate or prevent the flow of ions and organic osmolytes. In this study we use a linear force model of MS channel gating to determine the gating membrane tension (γ) and the gating area change (ΔA) associated with the energetics of MscS channel gating in giant spheroplasts and azolectin liposomes. Analysis of Boltzmann distribution functions describing the dependence of MscS channel gating on membrane tension indicated that the gating area change (ΔA) was the same for MscS channels recorded in both preparations. The comparison of the membrane tension (γ) gating the channel, however, showed a significant difference between the MscS channel activities in these two preparations.     

Analysis of searching movements of a leafminer parasitoid in a structured environment

ABSTRACT. Pnigalio soemius (Hymenoptera, Eulophidae) is a polyphagous ectoparasitoid of the apple leafminer Phyllonorycter cydo-niella (Lepidoptera, Gracillaridae). The tracks of females searching on the unmined leaf lamina and on the mine were segmented into 2 mm steps and analysed for angle distributions and correlations between successive steps. The movements on the mine are characterized by a higher mean and higher variance of turn angles compared to the turn angles on the leaf (kinesis-reaction family). Both klinokinesis and schemakinesis, for which a more precise definition is proposed, are present. Varying the step length has a great influence on the result and can be used to identify different orientation reactions within one track. The insect turns sharply once the mine border has been reached, a behaviour shown to be a crude taxis. An ecologically related species, Sympiesis sericeicornis (Hymenoptera, Eulophidae), moves in the same way. For this species a taxis towards the host could also be observed. A complex mixture of these orientation behaviours allows the parasitoid to remain on the mine and to localise the host.     

Patch occupancy of two hemipterans sharing a common host plant

Aim Two hemipteran species were chosen as a study system for the comparative analysis of patch occupancy and spatial population structure of insects sharing a common host plant. This study tested whether (1) the incidence in the host plant patches differed between the two species, and (2) the two species exhibited a different spatial population structure, i.e. were they affected differentially by isolation and area of the host plant patches. Location The porphyry landscape north of Halle (Saale) in Germany comprising 506 patches of the host plant Brachypodium pinnatum. Methods The host plant patches were surveyed for the two hemipterans. To assess the influence of patch quality on species occurrence the patches were characterized by mean cover abundance of B. pinnatum, type of subsoil, slope, exposure, and shading. The spatial configuration of the patches was considered by patch area and isolation. The influence of the habitat factors and the spatial configuration on the occupancy of the two species was analysed by logistic regression. Results Adarrus multinotatus was found in 441 patches, while Neophilaenus albipennis was found in only 90 patches. While A. multinotatus showed virtually no relationship to the habitat factors, the occupancy of N. albipennis was influenced by subsoil type, cover abundance, and shading. The effects of area and isolation on occupancy of the patches also differed between the two species. The occupancy of N. albipennis was determined largely by area and isolation, whereas in A. multinotatus no considerable effect of spatial configuration was found. Main conclusions The study revealed a marked difference between the two hemipteran species in respect of spatial population structure. Adarrus multinotatus built up a ‘patchy population’, whereas N. albipennis showed a ‘metapopulation’ structure within the same set of patches in the same landscape. Spatial population structure was found to be not only a function of spatial configuration of habitat patches, but population structure differed between the habitat generalist A. multinotatus and the habitat specialist N. albipennis.     

Host kairomone learning and foraging success in an egg parasitoid: a simulation model

Abstract 1. Trissolcus basalis (Wollaston) (Hymenoptera: Scelionidae) is an egg parasitoid that recognises chemical residues left by its host the green stink bug Nezara viridula (L.) (Heteroptera: Pentatomidae) as kairomone signals, enabling it to find egg masses in which to lay eggs. 2. Kairomones are usually present as patches deposited by N. viridula females, and recent results (Peri et al., Journal of Experimental Biology, 209 , 3629–3635, 2006) indicated that females of T. basalis are able to learn the features of their foraging environment and to adjust accordingly the amount of time spent on the patches of kairomones they are visiting, depending on whether or not host eggs are found. 3. In order to assess the impact of this learning ability, a Monte Carlo, spatially explicit and individual‐based simulation model was built to quantify the foraging efficiency of T. basalis females in environments with different levels of host abundance and distribution. In all cases, the present study compared the foraging efficiency of simulated T. basalis females having the ability to learn with those lacking this ability. 4. Learning females always visited a higher number of kairomone patches and attacked a higher number of hosts than non‐learning females, especially when there was a high density of kairomone patches in the environment. 5. Learning ability globally appears to allow the maintenance of efficient foraging success, especially when there is a low probability for the kairomone patches to contain discoverable hosts. 6. The increase in foraging efficiency for learning females appears to depend on the characteristics of the habitat in which they are foraging. Results thus suggest that significant variation in learning ability is likely to occur in natural wasp populations facing different environments with different host spatial distributions.