Structural Properties and Features of Parasitic Bdellovibrio bacteriovorus

The structure of five parasitic strains of Bdellovibrio bacteriovorus was studied by electron microscope after negative staining and in shadow-case and etched freeze-fractured preparations. Special attention was paid to the cell wall and the flagellar sheath which is continuous with the wall or part of it. These structural components reveal distinct features which are induced by certain staining substances; they are exceedingly susceptible to disruption by physical treatments, and in old cells often appear impaired. In freeze-fractured cells the wall shows characteristic fracturing tendencies not known in other microorganisms. These structural properties and features are distinct to Bdellovibrio wall and flagellar sheath, the structural integrity of which is a fundamental requirement for the infectivity and survival of this organism. The anterior end of Bdellovibrio is differentiated: 6 to 12 ring-like structures (9 to 12 nm, outer diameter) are built into its wall and several fibers (7 to 10 nm wide, up to 1.5 μm long) emerge from it. Intracellular structures, which are revealed as compact oval bodies bulging from the cell border and have internal laminated organization, are characteristic of Bdellovibrio after negative staining with certain compounds. These findings on the structure of parasitic Bdellovibrio substantiate previous observations indicating the uniqueness of this organism and add criteria for the identification of this genus.     

Lesions of the Basal Forebrain Cholinergic System in Mice Disrupt Idiothetic Navigation

Loss of integrity of the basal forebrain cholinergic neurons is a consistent feature of Alzheimer’s disease, and measurement of basal forebrain degeneration by magnetic resonance imaging is emerging as a sensitive diagnostic marker for prodromal disease. It is also known that Alzheimer’s disease patients perform poorly on both real space and computerized cued (allothetic) or uncued (idiothetic) recall navigation tasks. Although the hippocampus is required for allothetic navigation, lesions of this region only mildly affect idiothetic navigation. Here we tested the hypothesis that the cholinergic medial septo-hippocampal circuit is important for idiothetic navigation. Basal forebrain cholinergic neurons were selectively lesioned in mice using the toxin saporin conjugated to a basal forebrain cholinergic neuronal marker, the p75 neurotrophin receptor. Control animals were able to learn and remember spatial information when tested on a modified version of the passive place avoidance test where all extramaze cues were removed, and animals had to rely on idiothetic signals. However, the exploratory behaviour of mice with cholinergic basal forebrain lesions was highly disorganized during this test. By contrast, the lesioned animals performed no differently from controls in tasks involving contextual fear conditioning and spatial working memory (Y maze), and displayed no deficits in potentially confounding behaviours such as motor performance, anxiety, or disturbed sleep/wake cycles. These data suggest that the basal forebrain cholinergic system plays a specific role in idiothetic navigation, a modality that is impaired early in Alzheimer’s disease.     

The sodium current underlying action potentials in guinea pig hippocampal CA1 neurons

Neurons were acutely dissociated from the CA1 region of hippocampal slices from guinea pigs. Whole-cell recording techniques were used to record and control membrane potential. When the electrode contained KF, the average resting potential was about -40 mV and action potentials in cells at -80 mV (current-clamped) had an amplitude greater than 100 mV. Cells were voltage-clamped at 22-24 degrees C with electrodes containing CsF. Inward currents generated with depolarizing voltage pulses reversed close to the sodium equilibrium potential and could be completely blocked with tetrodotoxin (1 microM). The amplitude of these sodium currents was maximal at about -20 mV and the amplitude of the tail currents was linear with potential, which indicates that the channels were ohmic. The sodium conductance increased with depolarization in a range from -60 to 0 mV with an average half-maximum at about -40 mV. The decay of the currents was not exponential at potentials more positive than -20 mV. The time to peak and half-decay time of the currents varied with potential and temperature. Half of the channels were inactivated at a potential of -75 mV and inactivation was essentially complete at -40 to -30 mV. Recovery from inactivation was not exponential and the rate varied with potential. At lower temperatures, the amplitude of sodium currents decreased, their time course became longer, and half-maximal inactivation shifted to more negative potentials. In a small fraction of cells studied, sodium currents were much more rapid but the voltage dependence of activation and inactivation was very similar.     

Effects of Seedbed Management on Blast Development in Susceptible and Partially Resistant Rice Cultivars

Four rice cultivars, susceptible or partially resistant to Pyricularia grisea were evaluated for their apparent infection rates (r) and for terminal severity values in seedlings grown in seedbeds. Susceptibility of partially resistant cultivars decreased when seeded in wet seedbed compared to those grown in raised or upland seedbed. The degree of blast reduction due to flooding varied with rice genotype, seeding rate, and environmental conditions. Flooding was effective in managing seedling blast of partially resistant cultivars but not of susceptible cultivar. Use of a lower seed rate in comparison to a higher seed rate also led to reductions in r-values and terminal disease severities. However, in case of the susceptible cultivar IR50, effect of seed rate was more pronounced, when low amount and short duration of rainfall occurred. Flooding of wet beds with low seed rate further reduced the blast incidence. Results of this study suggest that with appropriate use of water, seed rate, and partial host resistance there are prospects for effective control of leaf blast in the tropics.     

Precocene causes male determination in the aphid Myzus persicae

When adult apterous viviparous females of Myzus persicae, reared in short night conditions at 21–23°C, are treated with the precocene analogue 6-methoxy-7-ethoxy-2,2-dimethylchromene, they deposit males towards the end of their reproductive lives. The first-born normal daughters of the treated females also deposit some males at various times in their reproductive lives. Karyotypic analysis was used to investigate the sequence of male and female embryos in the ovarioles of precociously metamorphosed aphids. The experiments support the hypothesis (Mittler et al., 1979) that juvenile hormone level controls the sex determination process in aphids. Since male aphids have an XO sex chromosome constitution, this implies that juvenile hormone level influences the behaviour of the X-chromosomes at or before the single maturation division of the egg. At this division one X-chromosome is eliminated from eggs which will develop as males. Aphids provide the first example of a specific endocrine influence on chromosome behaviour in sex determination.     

Penetration of Bdellovibrio bacteriovorus into Host Cells

Electron microscopy reveals that, in Bdellovibrio infection, after the formation of a passage pore in the host cell wall, the differentiated parasite penetration pole is associated with the host protoplast. This firm contact persists throughout the parasite penetration and after this process is completed. In penetrated hosts this contact is also apparent by phase microscopy. The association between the walls of the parasite and the host at the passage pore, on the other hand, is transient. Bdellovibrio do not penetrate hosts whose protoplast and cell walls are separated by plasmolysis, or in which the membrane-wall relationship is affected by low turgor pressure. It is concluded, therefore, that for penetration to occur it is essential that the host protoplast be within reach of the parasite, so that a firm contact can be established between them. A penetration mechanism is proposed that is effected by forces generated by fluxes of water and solutes due to structural changes in the infected host envelope. These forces cause a differential expansion of the host protoplast and cell wall and their separation from each other around the entry site, while the parasite remains firmly anchored to the host protoplast. Consequently, the parasite ends up enclosed in the expanded host periplasm. The actual entry, therefore, is a passive act of the parasite.     

Transduction of Merodiploidy: Induced Duplication of Recipient Genes

Escherichia coli PB160, which carries a tandem duplication with the gene order metB(+)argH(-)su(159) (+)thi(+): metB(+)argH(+)su(159) (-)thi(+), was used to study the mechanism of P1 transduction of genes in the duplicated region. Transduction of the su(159) (+) allele contained within the duplicated segment yields two kinds of su(159) (+) recombinants: 91% are haploid su(159) (+) and 9% are su(159) (+)/su(159) (-) merodiploids. The duplication in these merodiploid transductants includes the metB locus; however, both copies of the metB locus usually are derived from the recipient. Thus, the requirements for transduction of the "condition of merodiploidy" appear to be the cotransduction of the repeat point (the region where the duplication begins to repeat itself) and, of course, the selected marker (in this case su(159) (+)). A mechanism whereby two recipient chromosomes interact with the transduced "repeat point" region to regenerate the tandem duplication is implicated. It appears that a duplication much larger than the quantity of genetic material carried by a P1 phage can be produced in a transductant.     

Senescence Mutants of Saccharomyces Cerevisiae with a Defect in Telomere Replication Identify Three Additional Est Genes

The primary determinant for telomere replication is the enzyme telomerase, responsible for elongating the G-rich strand of the telomere. The only component of this enzyme that has been identified in Saccharomyces cerevisiae is the TLC1 gene, encoding the telomerase RNA subunit. However, a yeast strain defective for the EST1 gene exhibits the same phenotypes (progressively shorter telomeres and a senescence phenotype) as a strain deleted for TLC1, suggesting that EST1 encodes either a component of telomerase or some other factor essential for telomerase function. We designed a multitiered screen that led to the isolation of 22 mutants that display the same phenotypes as est1 and tlc1 mutant strains. These mutations mapped to four complementation groups: the previously identified EST1 gene and three additional genes, called EST2, EST3 and EST4. Cloning of the EST2 gene demonstrated that it encodes a large, extremely basic novel protein with no motifs that provide clues as to function. Epistasis analysis indicated that the four EST genes function in the same pathway for telomere replication as defined by the TLC1 gene, suggesting that the EST genes encode either components of telomerase or factors that positively regulate telomerase activity.