Chemotaxis in Physarum polycephalum : Effects of chemicals on isometric tension of the plasmodial strand in relation to chemotactic movement

The effects of chemicals were examined on the isometric tension of the plasmodial strand of the true slime mold Physarum polycephalum, and chemotactic motive forces were compared with the contractile properties of the strand. The results were:

1. 1. Isometric tension changed rhythmically within a period of 3–4 min and a few mg in amplitude. Application of attractants (glucose, galactose, maltose, Ca(H₂PO₄)₂, and K(H₂PO₄) led to a decrease in the amplitude of the tension. Contrary to this, repellents (sucrose, inorganic salts) increased the amplitude of the tension. The base line of the tension did not change appreciably unless the concentration of chemicals applied was not too high as compared with respective thresholds.

2. 2. Changes in the isometric tension, F, induced by application of chemicals were analysed quantitatively in terms of integral of isometric tension with respect to time during a period as defined by S=F dt. The values of S changed gradually with increase of concentration of chemicals above their respective thresholds.

3. 3. The threshold concentrations of chemicals determined by measurements of the isometric tension agreed with those obtained from chemotactic motive force and from membrane potential changes.

4. 4. The plasmodium of Physarum polycephalum moved away vigorously from high osmolarity by producing a large transient increase of motive force of the protoplasmic movement. Similarly, the isometric tension increased transiently with a high peak when the concentration of sugars and glycerol exceeded 0.2 M. The maximum tension was linearly proportional to the diameter of the strand.

These results indicate that contraction or relaxation of the plasma gel is the primary cause of the negative and positive chemotaxis in the slime molds.     

Chemotaxis toward carbohydrates and amino acids in Physarum polycephalum

A method is described for assaying chemotaxis in the acellular slime mold Physarum polycephalum. It consists of measuring the amount of plasmodium that moves on a strip of nitrocellulose membrane filter Millipore in response to a gradient of an attractant. Time course of chemotactic response of the slime mold is described. Different factors that affect chemotaxis in the slime mold such as: culture care and stage of growth of microplasmodia, substratum used for cell movement, nature of the gradient, effect of salts, pH and temperature are described. From concentration-response curves for different attractants several parameters of the chemotactic effect, such as threshold concentration, half maximal concentration, and maximal effective concentration can be determined. As a group, sugars are more effective chemotactic agents than amino acids. Glucose and galactose, which support the growth of the slime mold, are shown to have high positive chemotactic effect. 3-O-Methyl- -glucose and 2-deoxy- -glucose are two sugars that do not support growth but are very effective attractants. Conversely, fructose which supports slime mold growth is at best a weak attractant. The results support the view that the chemotactic effects of different sugars are not dependent on their growth-supporting value.     

Induction of cell fusion by a factor released by the cellular slime mold Polysphondylium pallidum

Heterokaryons and hybrid cells, which are extremely useful for research in cell biology, can be produced artificially by treating cells with either polyethylene glycol or certain inactivated viruses that alter the plasma membrane. We report here a novel cell-fusion inducing factor secreted by CK-8 strain cells of cellular slime mold Polysphondylium pallidum. Treatment of other strains or other species of cellular slime molds, such as NC-4 of Dictyostelium discoideum with the diluted fraction, containing molecules larger than 50 kDa, of the conditioned medium of CK-8 cell culture induces cell fusion at high frequency and produces multinucleated large cells. This cell fusion is inducible between cells of either a single strain or of two different strains of cellular slime molds.Abbreviations BSS Bonner's salt solution - CM conditioned medium - EDTA ethylenediaminetetraacetic acid - F2 fraction containing cell-fusion induction factor - Mr molecular mass     

A NATURALLY OCCURRING DEVELOPMENTAL SYNERGISM BETWEEN THE CELLULAR SLIME MOLD,DICTYOSTELIUM MUCOROIDES AND THE FUNGUS,MUCOR HIEMALIS

We here report the second record of a developmentally aberrant strain of a cellular slime mold from natural populations and demonstrate that this Dictyostelium mucoroides variant is capable of undergoing normal morphogenesis in the presence of the phycomycete fungus, Mucor hiemalis. The synergism is induced by an extracellular product(s) which is diffusable through thin agar membranes and is released by the fungus. The presence of the fungus not only induces stalk formation in this stalkless variant, but also increases the rate of sorocarp formation in 3 of 5 additional species of cellular slime molds assayed.     

Identification of the First Fungal Annexin: Analysis of Annexin Gene Duplications and Implications for Eukaryotic Evolution

Annexin homologues have been found in animals, plants, and distinct protist lineages. We report the identification of the first fungal annexin, encoded by the anx14 gene of the filamentous ascomycete Neurospora crassa. Annexins have a complex evolutionary history and exhibit a large number of gene duplications and gene losses in various taxa, including the complete loss of annexin sequences from another ascomycete, the budding yeast Saccharomyces cerevisiae. Surprisingly, the N. crassa annexin homologue is most closely related to the annexin homologue of the slime mold Dictyostelium discoideum, suggesting a phylogenetic link between cellular slime molds and true fungi. Both of these annexin homologues are closely related to the family of annexin homologues present in animals, an observation consistent with the existence of the animal–fungal clade. These data further suggest that the gene duplications that generated the family of annexin sequences present in animals, fungi, and slime molds began prior to the divergence of these taxa. Received: 10 December 1997 / Accepted: 17 April 1998     

Deoxyribonucleic Acid Base Composition of Some Cellular Slime Molds*

SYNOPSIS. Deoxyribonucleic acids (DNAs) isolated from 12 representatives of cellular slime molds grown in monoxenic cultures were analyzed by density gradient centrifugation in CsC1. The unique contribution of the DNA of each food organism was subtracted and the DNA base composition of each cellular slime mold determined. The guanine plus cytosine contents range 22–37 moles % within the representatives of the order. Minor bands (satellite bands) of DNA have been observed in preparations from Dictyostelium spp. and from Polysphondylium pallidum.     

THE OCCURRENCE AND DISTRIBUTION OF ACRASIEAE IN FOREST SOILS. I. EUROPE

The deciduous forests of Europe contain fewer species of cellular slime molds than those of Eastern North America. Diclyostelium mucoroides is the most widely distributed and the most abundant species. In Europe this species is as a rule the sole dominant species in a forest-soil acrasian population, a situation that rarely occurs in North America. An explanation of this difference may lie in the severity of the Pleistocene climate on European forests which resulted in the elimination of suitable habitats for the Acrasieae over a long period of time, and the replacement of the original forest by one much poorer in tree species. It is suggested that this modern forest does not offer the diverse of habitat which may be requited to support a mixed population of cellular slime molds.     

Ground-feeding migratory songbirds as cellular slime mold distribution vectors

Cosmopolitan species of cellular slime molds occur continents apart in both tropical and temperate zones of the world though the spore masses are too heavy to be wind borne, and water dispersal is limited to the watercourses. A highly mobile distribution vector was found in ground-feeding migratory song birds. Nine ubiquitous species and 2 ecologicially distinct species of dictyostelid cellular slime molds were isolated from the feces of ground-feeding eastern North American migratory thrushes, finches, sparrows and warblers, both on breeding and winter grounds. Three propagules of slime molds, amoebae, spores and macrocysts survive passage through the avian digesive tract and remain in the gut long enough to be transported by major bird migrations. Habitats with the greates species diversity of both cellular slime molds and ground-feeding passerines concur in both eastern North America and Central America. Birds actively seek their prefered habitats; the cellular slime molds have arrived at these habitats as passengers. Rare slime molds can serve as a marker to the habitats that migratory birds have visited, or birds with known habitats can provide clues as to the distribution of rare species of cellular slime molds.     

CELLULAR SLIME MOLDS OF SOUTHEAST ASIA. I. DESCRIPTION OF NEW SPECIES

Sixteen species of cellular slime molds were isolated from Southeast Asian forest soils. Ten of these, Dictyostelium mucoroides Brefeld, D. purpureum Olive, D. polycephalum Raper, D. lacteum van Tieghem, D. rhizopodium Raper and Fennell, D. lavandulum Raper and Fennell, D. vinaceo-fuscum Raper and Fennell, D. coeruleo-stipes Raper and Fennell, Polysphondylium violaceum Brefeld, and P. pallidum Olive have been previously described and are well-recognized species occurring in other parts of the world. Two, one in the genus Dictyostelium and one belonging to the family Guttulinaceae, are considered species by the author but have not been formally described. Four are described in this paper as new: Dictyostelium intermedium, D. multi-stipes, D. bifurcatum, and Acytostelium subglobosum. A new variety papilloideum of D. lacteum is also described. One other discovery of special interest is an isolate of Polysphondylium violaceum which produces abundant macrocysts, now known to be the sexual stage in the life cycle of cellular slime molds.     

Novel chlorinated dibenzofurans isolated from the cellular slime mold,Polysphondylium filamentosum,and their biological activities

Cellular slime molds are expected to have the huge potential for producing secondary metabolites including polyketides, and we have studied the diversity of secondary metabolites of cellular slime molds for their potential utilization as new biological resources for natural product chemistry. From the methanol extract of fruiting bodies of Polysphondylium filamentosum, we obtained new chlorinated benzofurans Pf-1 (4) and Pf-2 (5) which display multiple biological activities; these include stalk cell differentiation-inducing activity in the well-studied cellular slime mold, Dictyostelium discoideum, and inhibitory activities on cell proliferation in mammalian cells and gene expression in Drosophila melanogaster.     

Dictyostelium discoideum mitochondrial DNA encodes a NADH:Ubiquinone oxidoreductase subunit which is nuclear encoded in other eukaryotes

Complex I, a key component of the mitochondrial electron transport system, is thought to have evolved from at least two separate enzyme systems prior to the evolution of mitochondria from a bacterial endosymbiont, but the genes for one of the enzyme systems are thought to have subsequently been transferred to the nuclear DNA. We demonstrated that the cellular slime mold Dictyostelium discoideum retains the ancestral characteristic of having mitochondria encoding at least one gene (80-kDa subunit) that is nuclear encoded in other eukaryotes. This is consistent with the cellular slime molds of the family Dictyosteliaceae having diverged from other eukaryotes at an early stage prior to the loss of the mitochondrial gene in the lineage giving rise to plants and animals. The D. discoideum mitochondrially encoded 80-kDa subunit of complex I exhibits a twofold-higher mutation rate compared with the homologous chromosomal gene in other eukaryotes, making it the most divergent eukaryotic form of this protein.Correspondence to: K.L. Williams     

Breakdown of self/nonself recognition in cannibalistic strains of the predatory slime mold, Dictyostelium caveatum

Dictyostelium caveatum amebas feed upon both bacteria and the amebas of other cellular slime molds. The capacity to feed extensively upon other cellular slime molds is unique to D. caveatum amebas. They are able to phagocytose amebas larger than themselves by nibbling pieces of the cells until they are small enough to ingest. Here we report the isolation from previously cloned stock cultures of stable, cannibalistic strains of D. caveatum in which self/nonself recognition has broken down. Because of the extensive cannibalism, amebas of these strains do not complete multicellular development, and instead wander about for long periods while feeding upon each other. Although the cannibalistic behavior resembles that exhibited by the presumably diploid giant cells in the sexual cycle of other cellular slime molds, these strains are haploid and do not form macrocysts.     

Achene morphology and slime structure in some taxa of Artemisia L. and Neopallasia L. (Asteraceae)

We have examined slime cell distribution on the surface of the achenes of some Artemisia and Neopallasia taxa, as well as slime composition, envelope formation during the hydration, and slime relation to different morphological features and environmental factors. The results of the studies show a characteristic pattern of slime cells distribution, which could differ between taxa. The slime in the taxa studied belongs to the cellulose type and consists of two components i.e., pectins and cellulose. Although all fruits contain slime cells, not all of them show the slime envelope formation. Plants occurring in dry habitats (such as A. barrelieri) or annual species (such as A. annua) are characterised by a large amount of slime and a fast process of slime envelope formation. Slime production has not been observed in some polyploid populations (A. campestris and A. campestris ssp. sericea) and in two species occurring in relatively fertile habitats (A. verlotiorum, A. vulgaris). A reason for this may be either the immaturity of polyploid fruits leading to the production of a scarce, not detectable slime amount or, alternatively, the occurrence of not functional slime cells. Slime facilitates and stimulates the germination, as well as the adherence of the fruits to the ground or to animals (for dispersal). The slime could play important role in the distribution and colonisation of new habitats in many Artemisia taxa.     

THE YELLOW-PIGMENTED DICTYOSTELIA

The cellular slime molds described herein are of a clear golden yellow color and belong to two naturally occurring groups. One we call Dictyostelium mexicanum sp. n. because it is found in soils from various parts of tropical and subtropical Mexico. The well-tapered stalk and disklike base are somewhat reminiscent of D. discoideum. The other is regarded as D. aureum E. W. Olive (1901) with which we feel it is closely identified if not identical. In the latter group the form of both the aggregation and sorocarp is suggestive of D. mucoroides. Some less pigmented isolates are separated as D. aureum var. luteolum var. n.     

Regulation of aggregate size and pattern by adenosine and caffeine in cellular slime molds

Background

Multicellularity in cellular slime molds is achieved by aggregation of several hundreds to thousands of cells. In the model slime mold Dictyostelium discoideum, adenosine is known to increase the aggregate size and its antagonist caffeine reduces the aggregate size. However, it is not clear if the actions of adenosine and caffeine are evolutionarily conserved among other slime molds known to use structurally unrelated chemoattractants. We have examined how the known factors affecting aggregate size are modulated by adenosine and caffeine.

Result

Adenosine and caffeine induced the formation of large and small aggregates respectively, in evolutionarily distinct slime molds known to use diverse chemoattractants for their aggregation. Due to its genetic tractability, we chose D. discoideum to further investigate the factors affecting aggregate size. The changes in aggregate size are caused by the effect of the compounds on several parameters such as cell number and size, cell-cell adhesion, cAMP signal relay and cell counting mechanisms. While some of the effects of these two compounds are opposite to each other, interestingly, both compounds increase the intracellular glucose level and strengthen cell-cell adhesion. These compounds also inhibit the synthesis of cAMP phosphodiesterase (PdsA), weakening the relay of extracellular cAMP signal. Adenosine as well as caffeine rescue mutants impaired in stream formation (pde4 ^- and pdiA ^- ) and colony size (smlA ^- and ctnA ^- ) and restore their parental aggregate size.

Conclusion

Adenosine increased the cell division timings thereby making large number of cells available for aggregation and also it marginally increased the cell size contributing to large aggregate size. Reduced cell division rates and decreased cell size in the presence of caffeine makes the aggregates smaller than controls. Both the compounds altered the speed of the chemotactic amoebae causing a variation in aggregate size. Our data strongly suggests that cytosolic glucose and extracellular cAMP levels are the other major determinants regulating aggregate size and pattern. Importantly, the aggregation process is conserved among different lineages of cellular slime molds despite using unrelated signalling molecules for aggregation.     

COPROMYXELLA: A NEW GENUS OF ACRASIDAE

A new genus of cellular slime molds, Copromyxella, is described together with four species that comprise it. Since it resembles Zopf's Copromyxa more than any genus presently recognized, but differs from it in important characteristics such as the smaller dimensions of its cellular elements and the delicacy of its fructifications, the name Copromyxella is chosen for the new taxon. Four species are included: C. silvatica, C. filamentosa, C. spicata, and C. coralloides. All are characterized by small myxamoebae with lobose and potentially “explosive” pseudopodia and may assume limax forms in liquid media; all possess nuclei with centrally positioned nucleoli; all lack true contractile vacuoles (with a single possible exception); all aggregate without stream formation; and all form fructifications with no demarcation into stalks and sori. Taxonomically the genus belongs in the Acrasidae with Copromyxa, Guttulinopsis and Acrasis rosea rather than in the better known Dictyostelidae that includes Dictyostelium and Polysphondylium.     

A new Escherichia coli gene, fdrA, identified by suppression analysis of dominant negative FtsH mutations

An Escherichia coli membrane protein, FtsH, has been implicated in several cellular processes, including integration of membrane proteins, translocation of secreted proteins, and degradation of some unstable proteins. However, how it takes part in such diverse cellular events is largely unknown. We previously isolated dominant negative ftsH mutations and proposed that FtsH functions in association with some other cellular factor(s). To test this proposal we isolated multicopy suppressors of dominant negative ftsH mutations. One of the multicopy suppressor clones contained an N-terminally truncated version of a new gene that was designated fdrA. The FdrA fragment suppressed both of the phenotypes — increased abnormal translocation of a normally cytoplasmic domain of a model membrane protein and retardation of protein export — caused by dominant negative FtsH proteins. The intact fdrA gene (11.9 min on the chromosome) directed the synthesis of a 60 kDa protein in vitro.     

Cyclic AMP and calcium exchange in a cellular slime mold

Cyclic AMP is known to be an effective chemotactic agent for amebae of the cellular slime mold D. discoideum. A large amount of information from experiments on metazoa suggested that one cellular effect of cyclic AMP might be to alter the permeability of the cell membrane to Calcium or Sodium ions. On the basis of this information experiments were designed to test the effect of cyclic AMP on outflow of labeled Calcium or Sodium ions from amebae of D. discoideum. It was found that addition of cyclic AMP at 10^?4M resulted in a large increase of Ca⁴⁵ outflow from cells at the pre-aggregative or aggregative stage of development. No effect was found on Na²² outflow. It is suggested that this effect on Calcium permeability of the membrane is related to the chemotactic influence of ATP by some action on the contractile mechanism for ameboid movement. The phenomenon may be distinct from the enzyme inductive activity of cyclic AMP known for bacteria, and perhaps occurring in the cellular slime molds as well.     

Antagonists of chemoattractants reveal separate receptors for cAMP, folk acid and pterin in Dictyostelium

Adenosine 3′,5′-monophosphate (cAMP), folic acid and pterin are chemoattractants in the cellular slime molds. The cAMP analog, 3′-amino-cAMP, inhibits a chemotactic reaction to cAMP at a concentration at which the analog is chemotactically inactive. The antagonistic effect of 3′-amino-cAMP on the chemotactic activity of cAMP is competitive, which suggests that 3′-amino-cAMP antagonizes cAMP via the chemotactic receptor for cAMP. 3′-Amino-cAMP does not antagonize folic acid or pterin. The binding of folic acid to post-vegetative Dictyostelium discoideum cells is inhibited by low concentrations of 2-deamino-2-hydro folic acid (DAFA [7]). DAFA is neither chemotactically active, nor does it inhibit a chemotactic reaction to folic acid. This questions the involvement of the main folic acid cell surface-binding sites in the chemotactic response to folic acid. The pterin analog, 6-aminopterin, is an antagonist of pterin, but not of cAMP or folic acid. Our results show that cAMP, folic acid and pterin are detected by different receptors. Furthermore, they suggest that the antagonistic action of 3′-amino-cAMP and 6-aminopterin is localized in the signal transduction pathway at a step before the signals from the separate receptors have arrived at a single pathway.