The response of the tandem pore potassium channel TASK-3 (K2P9.1) to voltage : gating at the cytoplasmic mouth

Although the tandem pore potassium channel TASK-3 is thought to open and shut at its selectivity filter in response to changes of extracellular pH, it is currently unknown whether the channel also shows gating at its inner, cytoplasmic mouth through movements of membrane helices M2 and M4.We used two electrode voltage clamp and single channel recording to show that TASK-3 responds to voltage in a way that reveals such gating. In wild-type channels, Popen was very low at negative voltages, but increased with depolarisation. The effect of voltage was relatively weak and the gating charge small, ∼0.17.Mutants A237T (in M4) and N133A (in M2) increased Popen at a given voltage, increasing mean open time and the number of openings per burst. In addition, the relationship between Popen andvoltagewas shifted to lesspositive voltages. Mutation of putative hinge glycines (G117A, G231A), residues that are conserved throughout the tandem pore channel family, reduced Popen at a given voltage, shifting the relationship with voltage to a more positive potential range. None of these mutants substantially affected the response of the channel to extracellular acidification. We have used the results from single channel recording to develop a simple kinetic model to show how gating occurs through two classes of conformation change, with two routes out of the open state, as expected if gating occurs both at the selectivity filter and at its cytoplasmic mouth

Comparative Feeding Ecology of Sea Birds of a Tropical Oceanic Island

In 1963 and 1964, 800 food samples, regurgitated or dropped by captured birds, were collected on Christmas Island (Pacific Ocean) from Phaethon rubricauda, Puffinus nativitatis, Pterodroma alba, Sterna fuscata, Anous stolidus, Gygis alba, Anous tenuirostris and Procelsterna cerulea....https://elischolar.library.yale.edu/peabody_museum_natural_history_bulletin/1023/thumbnail.jp

The landscape ecological impact of afforestation on the British uplands and some initiatives to restore native woodland cover

The majority of forest cover in the British Uplands had been lost by the beginning of the Nineteenth Century, because of felling followed by overgrazing by sheep and deer. The situation remained unchanged until a government policy of afforestation, mainly by exotic conifers, after the First World War up to the present day. This paper analyses the distribution of these predominantly coniferous plantations, and shows how they occupy specific parts of upland landscapes in different zones throughout Britain. Whilst some landscapes are dominated by these new forests, elsewhere the blocks of trees are more localised. Although these forests virtually eliminate native ground vegetation, except in rides and unplanted land, the major negative impacts are at the landscape level. For example, drainage systems are altered and ancient cultural landscape patterns are destroyed. These impacts are summarised and possible ways of amelioration are discussed. By contrast, in recent years, a series of projects have been set up to restore native forest cover, as opposed to the extensive plantations of exotic species. Accordingly, the paper then provides three examples of such initiatives designed to restore native forests to otherwise bare landscapes, as well as setting them into a policy context. Whilst such projects cover a limited proportion of the British Uplands they nevertheless restore forest to landscapes at a local level

The selectivity, voltage-dependence and acid sensitivity of the tandem pore potassium channel TASK-1 : contributions of the pore domains

We have investigated the contribution to ionic selectivity of residues in the selectivity filter and pore helices of the P1 and P2 domains in the acid sensitive potassium channel TASK-1. We used site directed mutagenesis and electrophysiological studies, assisted by structural models built through computational methods. We have measured selectivity in channels expressed in Xenopus oocytes, using voltage clamp to measure shifts in reversal potential and current amplitudes when Rb+ or Na+ replaced extracellular K+. Both P1 and P2 contribute to selectivity, and most mutations, including mutation of residues in the triplets GYG and GFG in P1 and P2, made channels nonselective. We interpret the effects of these—and of other mutations—in terms of the way the pore is likely to be stabilised structurally. We show also that residues in the outer pore mouth contribute to selectivity in TASK-1. Mutations resulting in loss of selectivity (e.g. I94S, G95A) were associated with slowing of the response of channels to depolarisation. More important physiologically, pH sensitivity is also lost or altered by such mutations. Mutations that retained selectivity (e.g. I94L, I94V) also retained their response to acidification. It is likely that responses both to voltage and pH changes involve gating at the selectivity filter

Orai and TRPC channel characterization in FcεRI-mediated calcium signaling and mediator secretion in human mast cells.

Inappropriate activation of mast cells via the FcεRI receptor leads to the release of inflammatory mediators and symptoms of allergic disease. Calcium influx is a critical regulator of mast cell signaling and is required for exocytosis of preformed mediators and for synthesis of eicosanoids, cytokines and chemokines. Studies in rodent and human mast cells have identified Orai calcium channels as key contributors to FcεRI-initiated mediator release. However, until now the role of TRPC calcium channels in FcεRI-mediated human mast cell signaling has not been published. Here, we show evidence for the expression of Orai 1,2, and 3 and TRPC1 and 6 in primary human lung mast cells and the LAD2 human mast cell line but, we only find evidence of functional contribution of Orai and not TRPC channels to FcεRI-mediated calcium entry. Calcium imaging experiments, utilizing an Orai selective antagonist (Synta66) showed the contribution of Orai to FcεRI-mediated signaling in human mast cells. Although, the use of a TRPC3/6 selective antagonist and agonist (GSK-3503A and GSK-2934A, respectively) did not reveal evidence for TRPC6 contribution to FcεRI-mediated calcium signaling in human mast cells. Similarly, inactivation of STIM1-regulated TRPC1 in human mast cells (as tested by transfecting cells with STIM1-KK(684-685)EE - TRPC1 gating mutant) failed to alter FcεRI-mediated calcium signaling in LAD2 human mast cells. Mediator release assays confirm that FcεRI-mediated calcium influx through Orai is necessary for histamine and TNFα release but is differentially involved in the generation of cytokines and eicosanoids

Estimation and simulation of foraging trips in land-based marine predators

The behavior of colony-based marine predators is the focus of much research globally. Large telemetry and tracking data sets have been collected for this group of animals, and are accompanied by many empirical studies that seek to segment tracks in some useful way, as well as theoretical studies of optimal foraging strategies. However, relatively few studies have detailed statistical methods for inferring behaviors in central place foraging trips. In this paper we describe an approach based on hidden Markov models, which splits foraging trips into segments labeled as “outbound”, “search”, “forage”, and “inbound”. By structuring the hidden Markov model transition matrix appropriately, the model naturally handles the sequence of behaviors within a foraging trip. Additionally, by structuring the model in this way, we are able to develop realistic simulations from the fitted model. We demonstrate our approach on data from southern elephant seals (Mirounga leonina) tagged on Kerguelen Island in the Southern Ocean. We discuss the differences between our 4-state model and the widely used 2-state model, and the advantages and disadvantages of employing a more complex model

A multiplex marker set for microsatellite typing and sexing of sooty terns Onychoprion fuscatus

OBJECTIVES: Seabirds have suffered dramatic population declines in recent decades with one such species being the sooty tern Onychoprion fuscatus. An urgent call to re-assess their conservation status has been made given that some populations, such as the one on Ascension Island, South Atlantic, have declined by over 80% in three generations. Little is known about their population genetics, which would aid conservation management through understanding ecological processes and vulnerability to environmental change. We developed a multiplex microsatellite marker set for sooty terns including sex-typing markers to assist population genetics studies. RESULTS: Fifty microsatellite loci were isolated and tested in 23 individuals from Ascension Island. Thirty-one were polymorphic and displayed between 4 and 20 alleles. Three loci were Z-linked and two autosomal loci deviated from Hardy-Weinberg equilibrium. The remaining 26 autosomal loci together with three sex-typing makers were optimised in seven polymerase chain reaction plexes. These 26 highly polymorphic markers will be useful for understanding genetic structure of the Ascension Island population and the species as a whole. Combining these with recently developed microsatellite markers isolated from Indian Ocean birds will allow for assessment of global population structure and genetic diversity