POPULATION STRUCTURE AND KINSHIP IN POLISTES (HYMENOPTERA,VESPIDAE): AN ANALYSIS USING RIBOSOMAL DNA AND PROTEIN ELECTROPHORESIS

Six variable protein loci and one variable ribosomal DNA restriction site were used for an analysis of population structure in five species of Polistes from Texas. A sample-reuse algorithm was developed that estimated F_ST, F_IS, and ø (the coefficient of kinship) from probabilities of identity. Of the four species analyzed in detail only one, Polistes exclamans, had statistically significant values of F_ST. These values may reflect natural constraints on successful nesting for migrants of this species. Three of the four species had significant values of F_IS and three of the four species had significant values of ø. In many cases ø also differed from the expected value under haplodiploidy and random mating. Values of ø did not differ from expectations under haplodiploidy and local inbreeding. These results emphasize that theories of social behavior and evolution based on coefficients of kinship should include some explicit consideration of population structure.     

Stream flow and predation effects on the spatial dynamics of benthic invertebrates

Field experiments were carried out to determine the influence of predation and prey movements on the accumulation of prey in enclosures. Experimental enclosures permitted exchange of prey with the benthos, but not of the large, predatory larvae of the caddisfly, Plectrocnemia conspersa (Curtis). Unseasonally heavy rainfalls during the experiment resulted in high flows and enabled us to examine the effects of a major, abiotic disturbance on invertebrate spatial dynamics. Prey colonization rates of cages without predators were determined in nine 24 h periods. Colonization rates increased exponentially with flow and were species-specific, depending on dispersal behaviour. Prey accumulation and predator impacts were measured in cages, with and without P. conspersa larvae, placed in the stream for 1, 2 or 3 weeks. Prey densities in cages increased with exposure time, but increases were not gradual and depended on flow regime. Flow was reduced within cages and they accumulated large numbers of invertebrates during high discharge. Analogous, naturally occurring refugia in the stream channel could be important for the recovery of lotic communities after major disturbances. Overall, prey densities were lowest in cages with predators. For fast colonizers, predation effects were detectable early in the experiment, but quickly obscured thereafter by continuous exchange of prey. For slow colonists, predation effects were detectable later, but persisted longer. Consumption rates for P. conspersa varied with prey density and flow regime. We suggest that the spatial dynamics of benthic invertebrates, especially as they are influenced by stochastic events, are important in understanding and detecting predation effects in stream communities.     

Isolation of a high specific activity pink, monomeric nitrous oxide reductase from Achromobacter cycloclastes

Nitrous oxide reductase from Achromobacter cycloclastes has been purified to homogeneity under aerobic conditions via DEAE-cellulose, phenyl-Sepharose, hydroxyapatite, and Sephacryl S-200 chromatography. It consists of a single polypeptide of MW 72 kDa, and contains 3.8 +/- 0.1 copper atoms per molecule. The enzyme is pink as isolated, yet exhibits a specific activity (86 U/mg) that is ca. 40 times greater than that observed for other N2O reductases under similar conditions. Double integration of the anomalous EPR spectrum at 77K showed the presence of 2.0 +/- 0.1 spins per molecule, implying the presence of EPR-silent copper atoms and/or spin-coupled mixed-valent centers.     

Growth of Octopine-Catabolizing Pseudomonas spp. under Octopine Limitation in Chemostats and Their Potential To Compete with Agrobacterium tumefaciens

The growth characteristics of five octopine-catabolizing pseudomonads have been determined in batch and continuous cultures. All five strains belonged to rRNA homology group I and showed a more psychrotrophic growth pattern than did Agrobacterium tumefaciens B6 and ATCC 15955. In chemostats limited by octopine, either as the source of carbon and nitrogen or the sole source of nitrogen, maximum specific growth rates and substrate affinities were lower than those in chemostats limited by glutamate. These growth dynamics were similar to those observed for Agrobacterium strains B6 and ATCC 15955 even though the catabolic genes and pathways are believed to be different in the two genera. An analysis of the yields in octopine-limited chemostats indicated that the use of octopine as the sole source of carbon and nitrogen was grossly inefficient. Octopine and presumably lysopine and octopinic acid provided a better source of nitrogen than of carbon. One of the Pseudomonas fluorescens strains, E175D, was able to produce its highest yield on octopine as a nitrogen source. Competition models formulated on pure culture parameters indicated that two of the Pseudomonas spp. would dominate A. tumefaciens B6 and ATCC 15955 when in simple competition for octopine as a limiting substrate.     

Ontogeny of an Extracellular Matrix Component of Sea Urchins and its Role in Morphogenesis

A monoclonal antibody, Sp14, recognizes fibers that form a complex meshwork within the blastocoel of embryos of the sea urchin, Strongylocentrotus purpuratus . The fibers first appear as the blastocoel begins to form and increase in density throughout development. Ultrastructural localizations using the immunoperoxidase method show bundles of 20 nm fibers that are continuous with the basal lamina and have an indistinct axial periodicity. Embryos treated with tunicamycin, β-D-xylopyranoside, β-aminoproprionitrile, proline analogues, or deprived of sulfate all form immunoreactive fibers although in some treatments the pattern formed is abnormal. Immunoreactivity of extracted fibers is not affected by digestion with chondroitinase ABC, hyaluronidase, collagenase or heparinase. However, proteinase K readily destroys immunoreactivity. Fibers will form in cultures of micromeres or mesenchyme 24 to 48 hr after plating with or without horse serum. In embryos in which the blastocoelar matrix has been altered by injection with Sp14, there is inhibition of the release of secondary mesenchyme from the tip of the archenteron and in some embryos supernumerary skeletal elements are formed. It is proposed that Sp14 recognizes a component of the blastocoelar extracellular matrix that is required for the migration of mesenchyme.     

Measurement of release of endogenous GABA and catabolites of [3H]GABA from synaptosomal preparations using ion-exchange chromatography

Picomole quantities of endogenous GABA in acidified superfusates of synaptosomal preparations have been measured using micro-bore ion-exchange chromatography and post-column formation of the fluorescent iso-indole derivative. Using this technique superfusates have been analyzed directly, without further manipulations, to investigate the release of endogenous GABA. Spontaneous release of GABA was 2–5 pmol/200 l superfusate increasing to 20 pmol/200 l with potassium stimulation. When -vinyl GABA (RMI 71754), an inhibitor of GABA-T was injected into rats (750 mg/kg) and synaptosomes prepared the potassium-evoked release of GABA was increased 3-fold compared to controls. Chromatographic separations and measurement of release of endogenous and radiolabeled GABA allowed the real specific activity of released GABA to be calculated. Only when 500 M amino-oxyacetic acid was added during isolation of synaptosomes was the specific activity of released GABA the same as the initial specific activity.     

Presence of Radiolabelled Metabolites in Release Studies Using [3H]γ-Aminobutyric Acid

Abstract: Most studies on γ-aminobutyric acid (GABA) release from nervous tissue have been conducted using radiolabelled GABA in the presence of aminooxyacetic acid (AOAA) to inhibit GABA: 2-oxoglutarate aminotransferase (GABA-T) to prevent conversion of labelled GABA to labeled catabolites. Here we present data showing that even in the presence of 10 μM-AOAA the spontaneous release of tritium from rat cortical synaptosomes prelabelled with 2,3-[³H]GABA is mainly in the form of tritiated water but that the increase in tritium release in the presence of unlabelled GABA or high potassium-ion concentrations is in the form of authentic [³H]GABA. Interpretation of results should take these facts into account.