Transduction of the chemotactic signal to the actin cytoskeleton of Dictyostelium discoideum

Dictyostelium discoideum amebae chemotax toward folate during vegetative growth and toward extracellular cAMP during the aggregation phase that follows starvation. Stimulation of starving amebae with extracellular cAMP leads to both actin polymerization and pseudopod extension (Hall et al., 1988, J. Cell. Biochem. 37, 285-299). We have identified an actin nucleation activity (NA) from starving amebae that is regulated by cAMP receptors and controls actin polymerization (Hall et al., 1989, J. Cell Biol., in press). We show here that NA from vegetative cells is also regulated by chemotactic receptors for folate. Our studies indicate that NA is an essential effector in control of the actin cytoskeleton by chemotactic receptors. Guided by a recently proposed model for signal transduction from the cAMP receptor (Snaar-Jagalska et al., 1988, Dev. Genet. 9, 215-225), we investigated which of three signaling pathways activates the NA effector. Treatment of whole cells with a commercial pertussis toxin preparation (PT) inhibited cAMP-stimulated NA. However, endotoxin contamination of the PT appears to account for this effect. The synag7 mutation and caffeine treatment do not inhibit activation of NA by cAMP. Thus, neither activation of adenylate cyclase nor a G protein sensitive to PT treatment of whole cells is necessary for the NA response. Actin nucleation activity stimulated with folate is normal in vegetative fgdA cells. However, cAMP suppresses rather than activates NA in starving fgdA cells. This indicates that the components of the actin nucleation effector are present and that a pathway regulating the inhibitor(s) of nucleation remains functional in starving fgdA cells. The locus of the fgdA defect, a G protein implicated in phospholipase C activation, is directly or indirectly responsible for transduction of the stimulatory chemotactic signal from cAMP receptors to the nucleation effector in Dictyostelium.     

Cloning of an Aeromonas hydrophila type IV pilus biogenesis gene cluster: complementation of pilus assembly functions and characterization of a type IV leader peptidase/N-methyltransferase required for extracellular protein secretion

Aeromonas hydrophila secretes several extracellular proteins that are associated with virulence including an enterotoxin, a protease, and the hole-forming toxin, aerolysin. These degradative enzymes and toxins are exported by a conserved pathway found in many Gram-negative bacteria. In Pseudomonas aeruginosa this export pathway and type IV pilus biogenesis are dependent on the product of the pilD gene. PilD is a bifunctional enzyme that processes components of the extracellular secretory pathway as well as a type IV prepilin. An A. hydrophila genomic library was transferred into a P. aeruginosa pilD mutant that is defective for type IV pilus biogenesis. The A. hydrophila pilD homologue, tapD , was identified by its ability to complement the pilD mutation in P. aeruginosa . Transconjugants containing tapD were sensitive to the type IV pilus-specific phage, PO4. Sequence data revealed that tapD is part of a cluster of genes ( tapABCD ) that are homologous to P. aeruginosa type IV pilus biogenesis genes ( pilABCD ). We showed that TapB and TapC are functionally homologous to P. aeruginosa PilB and PilC, the first such functional complementation of pilus assembly demonstrated between bacteria that express type IV pili. In vitro studies revealed that TapD has both endopeptidase and N -methyltransferase activities using P. aeruginosa prepilin as substrate. Furthermore, we show that tapD is required for extracellular secretion of aerolysin and protease, indicating that tapD may play an important role in the virulence of A. hydrophila     

Geochemical partitioning and bioavailability of copper to aquatic plants in an artificial oxide-organic sediment

This study investigated the effects of competition between binding substrates (organic matter and iron oxide) and between metals (cadmium and copper), on the partitioning of sedimentary copper and its subsequent bioavailability to an aquatic plant. Organic matter and a synthesized iron oxide, ferrihydrite, were added singly and in combination to a series of sand sediments, which were then dosed with environmentally realistic concentrations of cadmium and copper and planted with rice,Oryza sativa. Organic matter controlled copper partitioning and bioavailability, whereas the synthetic ferrihydrite bound negligible amounts of either metal, even in the absence of organic matter. As organic matter concentrations increased, operationally-defined leachable copper decreased, organic-associated copper increased and the survival of rice plants improved in an approximately linear fashion. At a nominal starting copper concentration of 5.8 μg g dry wt⁻¹, plant survival after four weeks averaged 0–8% in sediments without organic matter, 25% in a sediment containing 0.18% organic matter and 58% in a sediment containing 0.36% organic matter. These results suggest that organic-associated forms of copper are unavailable to plants, and that the operational definition of ‘leachable’ copper (extracted with dilute ammonium acetate) adequately represents the species of copper that is (are) available to plants. Our study using a well-characterized artificial sediment supports the copper fractionation patterns and correlations between copper partitioning and bioavailability reported from the heterogeneous, poorly characterized sediments of natural lake and river sediments.     

The transposable element mariner can excise in non-drosophilid insects

Plasmid-based excision assays performed in embryos of two non-drosophilid species using the mariner transposable element from Drosophila mauritiana resulted in empty excision sites identical to those observed after the excision of mariner from D. mauritiana chromosomes. In the presence of the autonomous mariner element Mos1, excision products were recovered from D. melanogaster, D. mauritiana and the blowfly Lucilia cuprina. When a hsp82 heat shock promoter-Mos1 construct was used to supply mariner transposase, excision products were also recovered from the Queensland fruitfly Bactrocera tryoni. Analysis of DNA sequences at empty excision sites led us to hypothesise that the mariner excision/repair process involves the formation of a heteroduplex at the excision breakpoint. The success of these assays suggests that they will provide a valuable tool for assessing the ability of mariner and mariner-like elements to function in non-drosophilid insects and for investigating the basic mechanisms of mariner excision and repair.     

Habitat use and support preference of two free-ranging saltatory lemurs (Lepilemur edwardsi and Avahi occidentalis)

Lepilemur edwardsi and Avahi occidentalis are two species of nocturnal, folivorous 'vertical clingers and leapers' (VCL). They have a similar body mass and share the same morphological adaptation for leaping. In a field study under sympatric conditions at Ampijoroa, Madagascar, comparison of support use with support availability using Jacobs' D preference values (Jacobs, 1974) showed that both species actively chose or avoided branches with certain qualities. However, while both species showed a preference for small oblique and horizontal branches, they selected them at different heights in the forest and with varying degrees of preference and avoidance for the other available supports. Despite their traditional locomotor assignation, both species showed a surprisingly strong preference for horizontal supports. These striking variations in detail of support preference may aid the maintenance of species segregation and niche difference.     

The Cohesion-Tension theory of sap ascent: current controversies

In recent years, the Cohesion-Tension (C-T) theory of sap ascentin plants has come under question because of work publishedby Professor Ulrich Zimmermann and colleagues at the Universityof Wrzburg, Germany. The purpose of this review is to (1) statethe essential and testable elements of the C-T theory, (2) summarizethe negative evidence for the C-T theory, and (3) review criticallythe positive evidence for the C-T theory and the evidence thatthe Scholander-Hammel pressure bomb measures xylem pressurepotential (P_x) correctly, because much of the evidence for theC-T theory depends on pressure bomb data. Much of the current evidence negates the conclusions drawn byZimmermann from studies using the xylem pressure probe (XPP),but it is not yet clear in every instance why the XPP resultsdisagree with those of other methods for estimating xylem pressure.There is no reason to reject the XPP as a useful new tool forstudying xylem tensions in the range of 0 to –0.6 MPa.Additional research is needed to test the C-T theory with boththe XPP and traditional methods. Key words: Cohesion-Tension theory, cavitation, embolism, xylem pressure probe, pressure bomb     

Carbohydrate and the cytokine response to 2.5h of running

Nehlsen-Cannarella, S. L., O. R. Fagoaga, D. C. Nieman, D. A. Henson, D. E. Butterworth, R. L. Schmitt, E. M. Bailey, B. J. Warren, A. Utter, and J. M. Davis. Carbohydrate and the cytokineresponse to 2.5 h of running. J. Appl.Physiol. 82(5): 1662-1667, 1997.This randomized,double-blind, placebo-controlled study was designed to determine theinfluence of 6% carbohydrate (C) vs. placebo (P) beverage ingestion oncytokine responses (5 total samples over 9 h) to 2.5 h ofhigh-intensity running (76.7 ± 0.4% maximalO₂ uptake) by 30 experiencedmarathon runners. For interleukin-6 (IL-6), a difference in the patternof change between groups was found, highlighted by a greater increasein P vs. C immediately postrun (753 vs. 421%) and 1.5 h postrun (193 vs. 86%) [F(4,112) = 3.77, P = 0.006]. Forinterleukin-1-receptor antagonist (IL-1ra), a difference in the patternof change between groups was found, highlighted by a greater increasein P vs. C 1.5 h postrun (231 vs. 72%)[F(2,50) = 6.38, P = 0.003]. No significant interaction effects were seen for bioactive IL-6 or IL-1. The immediate postrun plasma glucose concentrations correlated negatively with those of plasma cortisol (r = 0.67, P < 0.001); postrun plasma cortisol (r = 0.70, P < 0.001) and IL-6 levels(r = 0.54, P = 0.003) correlated positively withlevels of IL-1ra. Taken together, the data indicate that carbohydrateingestion attenuates cytokine levels in the inflammatory cascade inresponse to heavy exertion.

     

Evidence for Linkage of Bipolar Disorder to Chromosome 18 with a Parent-of-Origin Effect

A susceptibility gene on chromosome 18 and a parent-of-origin effect have been suggested for bipolar affective disorder (BPAD). We have studied 28 nuclear families selected for apparent unilineal transmission of the BPAD phenotype, by using 31 polymorphic markers spanning chromosome 18. Evidence for linkage was tested with affected-sib-pair and LOD score methods under two definitions of the affected phenotype. The affected-sib-pair analyses indicated excess allele sharing for markers on 18p within the region reported previously. The greatest sharing was at D18S37: 64% in bipolar and recurrent unipolar (RUP) sib pairs (P = .0006). In addition, excess sharing of the paternally, but not maternally, transmitted alleles was observed at three markers on 18q: at D18S41, 51 bipolar and RUP sib pairs were concordant for paternally transmitted alleles, and 21 pairs were discordant (P = .0004). The evidence for linkage to loci on both 18p and 18q was strongest in the 11 paternal pedigrees, i.e., those in which the father or one of the father's sibs is affected. In these pedigrees, the greatest allele sharing (81%; P = .00002) and the highest LOD score (3.51; θ = 0.0) were observed at D18S41. Our results provide further support for linkage of BPAD to chromosome 18 and the first molecular evidence for a parent-of-origin effect operating in this disorder. The number of loci involved, and their precise location, require further study.     

Establishment and characterization of a new,spontaneously immortalized,pancreatic ductal cell line from the Syrian golden hamster

Spontaneously immortal pancreatic cell lines are not available. By use of a defined culture medium, such a line (TAKA-1) was established from the Syrian golden hamster. Cytological, cytogenetic, molecular biological, enzymatic and receptor patterns as well as antigenicity were studied and were compared with those of the normal hamster pancreatic ductal cells in vivo. TAKA-1 cells grew exponentially in a monolayer on collagen gel in a defined medium but did not proliferate in soft agar. Ultrastructurally, the cells closely resembled the normal hamster pancreatic ductal cells. Similarities and dissimilarities were found between the normal ductal cells and TAKA-1 cells. Similarities included the presence of cytokeratin, carbonic anhydrase and some tumor-associated antigens. However, unlike the normal ductal cells, TAKA-1 cells expressed blood group A angigen and anti-vimentin, showed affinity to selected lectins, and an abnormality of chromosome 3, which is suggested to be associated with immortality. Moreover, unlike the hamster pancreatic ductal cancer cells but like the normal hamster pancreatic ductal cells, TAKA-1 cells did not have a c-Ki-ras mutation. EGF, TGF- and secretin, but not CCK or GRP, bound to the TAKA-1 cells. TAKA-1 cells produced TGF-, and their growth was stimulated by exogenous EGF in serum-free medium. This cell line presents a suitable model for biologic and pathologic study of the hamster pancreatic ductal cells in vitro.