Further characterisation of the [35S]-TBPS binding site of the GABA receptor complex in locust (Schistocerca gregaria) ganglia membranes

1. Using homogenates of supraoesophageal ganglia from locust we observed specific binding of [35S]-TBPS which was linear with protein concentration up to 7 mg/ml, showed a pH optimum at pH 9.0 and was linear with NaCl concentration up to 350 mM. 2. Kinetic analysis of the binding showed positive cooperativity as a result of changes in on and off-rates with occupation of the binding site by the ligand. The apparent KD = 417 nM and Bmax = 1083 fmol/mg of membrane protein were calculated using a computer program for dose-response curve fitting. 3. The binding was enhanced by GABA, pentobarbital and benzodiazepines. Picrotoxinin had no effect on the binding at 0.1 mM. Only the cage convulsants TBPS and IBP were able to displace the binding. 4. Whilst the characteristics of the binding are similar to those reported for house fly thorax and abdomen preparations they are significantly different from those reported for house fly head, cockroach nerve cord and rat brain.     

Behavioural Reproductive Isolation in a Rotifer Hybrid Zone

A hybrid zone between two Brachionus plicatilis rotifer mitochondrial DNA (mtDNA) lineages was recently described in the Iberian Peninsula between a pond (Santed 2) and a lake (Gallocanta). The patterns of mitochondrial and nuclear genetic variation observed suggested that gene flow is mainly male-mediated from the lake to the pond. Here we test two hypotheses: (a) that male-mediated gene flow occurs through assortative mating between individuals from these ponds, (b) that behavioural isolation occurs between the two mtDNA lineages. We isolated, reared and genotyped rotifer clones from resting eggs collected in the sediments of these and two other distant ponds. We devised a quick, inexpensive RFLP method to discriminate between B. plicatilis and its sibling species B. ‘Manjavaeas’ and between both mtDNA B. plicatilis lineages. Behavioural no-choice tests using new-born, virgin males and females were performed between five clones. B. ‘Manjavacas’ and B. plicatilis were reproductively isolated. B. plicatilis clones did not show evidence of reproductive isolation, regardless of their mtDNA lineage, except Santed 2 males, which discriminated strongly against Gallocanta females. These results could help to explain the discrepancies between mitochondrial and nuclear genetic variation reported in the two populations.     

INCREASED INCORPORATION OF [G-3H]LEUCINE INTO A POSSIBLE ''RECEPTOR'' PROTEOLIPID IN DENERVATED MUSCLE IN VIVO

Abstract— The incorporation of radioactive leucine into the total proteins and the proteolipids of normal and denervated rat diaphragm has been studied in vivo. Denervation increased the incorporation of isotopically labelled leucine into each of the isolated proteolipids and the effect was particularly marked in a single proteolipid which has been designated a 'receptor' proteolipid. In normal muscle this particular proteolipid was found to have a higher incorporation of isotopically labelled leucine in the area of the muscle rich in endplates compared with an area devoid of endplates. However the stimulatory effect of denervation on the incorporation of radioactive leucine into this proteolipid was considerably more marked in the latter region. An attempt has been made to correlate these findings with the development of the hypersensitivity to ACh characteristic of denervated muscle.     

Spatial analysis of anthropogenic disturbance regimes and roadside shrubs in a fragmented agricultural landscape

Abstract. In fragmented woodlands in agricultural regions of Australia, roadside environments (road verges) provide important refuges for threatened native fauna and isolated populations of plant species. However, as roads are transport corridors for humans and their vehicles, species survival is affected through destruction, fragmentation and modification of remaining habitat by human activity. Few studies have recognised the effects of anthropogenic disturbances, in the form of historical roadworks activities, on adjacent roadside plant populations. The aim of this study was to investigate the effects of soil disturbance from roadworks on the spatial patterns and structural dynamics of roadside Acacia populations in an agricultural area in southeastern Australia. Stem size and disturbance data were collected from 135 roadside populations of three species of Acacia shrubs in the Lockhart Shire area. Spatial pattern analysis using the Network K‐function and Discriminant Function analyses showed that road verge width, road category, disturbance intensity and distance to nearest town were highly significant variables in relation to disturbance from roadworks and shrub structural dynamics. Colonising populations were more abundant along gravel roads where soil disturbance intensity was high, whereas stable populations were more abundant where soil disturbance intensity was low. Senescent populations were more abundant in narrow, little used road verge corridors 4–6 km from nearby towns. These results suggest that anthropogenic disturbance regimes may be critical to shrub recruitment and persistence in roadside environments, which has important consequences for conservation of threatened native flora and fauna in agricultural landscapes.     

A new global biome reconstruction and data-model comparison for the Middle Pliocene

Aim To produce a robust, comprehensive global biome reconstruction for the Middle Pliocene (c. 3.6–2.6 Ma), which is based on an internally consistent palaeobotanical data set and a state‐of‐the‐art coupled climate–vegetation model. The reconstruction gives a more rigorous picture of climate and environmental change during the Middle Pliocene and provides a new boundary condition for future general circulation model (GCM) studies. Location Global. Methods Compilation of Middle Pliocene vegetation data from 202 marine and terrestrial sites into the comprehensive GIS data base TEVIS (Tertiary Environmental Information System). Translation into an internally consistent classification scheme using 28 biomes. Comparison and synthesis of vegetation reconstruction from palaeodata with the outputs of the mechanistically based BIOME4 model forced by climatology derived from the HadAM3 GCM. Results The model results compare favourably with available palaeodata and highlight the importance of employing vegetation–climate feedbacks and the anomaly method in biome models. Both the vegetation reconstruction from palaeobotanical data and the BIOME4 prediction indicate a general warmer and moister climate for the Middle Pliocene. Evergreen taiga as well as temperate forest and grassland shifted northward, resulting in much reduced tundra vegetation. Warm‐temperate forests (with subtropical taxa) spread in mid and eastern Europe and tropical savannas and woodland expanded in Africa and Australia at the expense of deserts. Discrepancies which occurred between data reconstruction and model simulation can be related to: (1) poor spatial model resolution and data coverage; (2) uncertainties in delimiting biomes using climate parameters; or (3) uncertainties in model physics and/or geological boundary conditions. Main conclusions The new global biome reconstruction combines vegetation reconstruction from palaeobotanical proxies with model simulations. It is an important contribution to the further understanding of climate and vegetation changes during the Middle Pliocene warm interval and will enhance our knowledge about how vegetation may change in the future.     

Genetic tests of ancient asexuality in Root Knot Nematodes reveal recent hybrid origins

Background  

The existence of "ancient asexuals", taxa that have persisted for long periods of evolutionary history without sexual recombination, is both controversial and important for our understanding of the evolution and maintenance of sexual reproduction. A lack of sex has consequences not only for the ecology of the asexual organism but also for its genome. Several genetic signatures are predicted from long-term asexual (apomictic) reproduction including (i) large "allelic" sequence divergence (ii) lack of phylogenetic clustering of "alleles" within morphological species and (iii) decay and loss of genes specific to meiosis and sexual reproduction. These genetic signatures can be hard to assess since it is difficult to demonstrate the allelic nature of very divergent sequences, divergence levels may be complicated by processes such as inter-specific hybridization, and genes may have secondary roles unrelated to sexual reproduction. Apomictic species of Meloidogyne root knot nematodes have been suggested previously to be ancient asexuals. Their relatives reproduce by meiotic parthenogenesis or facultative sexuality, which in combination with the abundance of nematode genomic sequence data, makes them a powerful system in which to study the consequences of reproductive mode on genomic divergence.     

In Vivo Evidence for Serine Biosynthesis-Defined Sensitivity of Lung Metastasis,but Not of Primary Breast Tumors,to mTORC1 Inhibition

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Interrelationship of carbohydrate and the alpha-toxin binding site on the acetylcholine receptor from Torpedo marmorata

The relationship of the carbohydrate components of purified acetylcholine receptor (AChR) to its acetylcholine binding site was investigated by two approaches. In the first, the effect of periodate or glycosidase treatment of AChR on its ability to bind α-bungarotoxin was assessed. Although loss of binding capacity was observed, this could be attributed to increased temperature, acid pH or high salt concentrations of the incubation conditions rather than to the specific action of periodate or glycosidases, indicating that the α-toxin binding site does not directly involve carbohydrate.The second approach involved the use of concanavalin A to block the binding of α-toxin to AChR, when a maximum inhibition of approximately 40% was obtained. The results are interpreted in terms of heterogeneity of AChR molecules, of which some 40% have sterically interacting sites binding concanavalin A and α-toxin respectively.