Regular oscillations in suspensions of a putatively chaotic mutant ofDictyostelium discoideum

Summary We have tested the light-scattering properties of suspensions of theDictyostelium discoideum mutantHH201 derived from the mutantFr17. Previous studies indicated thatHH201 andFr17 possess highly irregular rhythmic properties which might represent aperiodic oscillations, i.e. chaos. We report that the former mutant can display regular oscillatory behavior. Possible explanations for this result are discussed, including that of a transition from chaotic to periodic behavior resulting from some parameter change or from strong intercellular coupling in cell suspensions.     

The histones ofDictyostelium discoideum

Summary The histones of the cellular slime mouldDictyostelium discoideum have been separated by electrophoresis using both acid urea and sodium dodecyl sulphate systems, and the gel pattern compared with that of histones fromPhysarum polycephalum and calf thymus.Dictyostelium is found to possess a full complement of H1, H2A, H2B, H3 and H4.     

Suppression of chaos by periodic oscillations in a model for cyclic AMP signalling inDictyostelium cells

We investigate how the introduction of cells oscillating periodically affects the behaviour of a suspension ofDictyostelium discoideum amoebae undergoing chaotic oscillations of cyclic AMP. The analysis of a model indicates that a tiny proportion of periodic cells suffices to transform chaos into periodic oscillations in such suspensions. A similar result is obtained by forcing the aperiodic oscillations by a small-amplitude, periodic input of cyclic AMP. The results provide an explanation for the observation of regular oscillations in suspensions of a putatively chaotic mutant ofDictyostelium discoideum ¹². More generally, the results show how chaos in biological systems may disappear through the coupling with periodic oscillations.     

Cell adhesion in the life cycle ofDictyostelium

three forms of cell adhesion determine the life cycle ofDictyostelium: i) adhesion of bacteria to the surface of the growing amoebae, as the prerequisite for phagocytosis;ii) cell-substrate adhesion, necessary for both locomotion of the amoebae and migration of the slug;iii) cell-cell adhesion, essential for transition from the unicellular to the multicellular stage. Intercellular adhesion has received the most attention, and fruitful approaches have been developed over the past 25 years to identify, purify and characterize cell adhesion molecules. The csA glycoprotein, in particular, which mediates adhesion during the aggregation stage, is one of the best defined cell adhesion molecules. The molecular components involved in phagocytosis and cell-substratum adhesion are less well understood, but the basis has been laid for a systematic investigation of both topics in the near future.     

Endocytosis and inositol hexakisphosphate levels inras transformants ofDictyostelium discoideum amoebae

Summary Fluid-phase pinocytosis kinetics and lysosomal enzyme secretion parameters were measured inDictyostelium discoideum amoebae constructed from strain AX3 by transformation with a multicopy plasmid carrying either a normalras gene (ras-Gly12), a mutatedras gene (ras-Thr12) or by the vector carrying the geneticin resistance gene only (pDNEO2). It was found that the pinocytosis rate and extent as well as the lysosomal enzyme secretion were slightly different in the three strains. These changes, however, were related to minor modifications of the cellular volumes. The overall concentration of inositol hexakisphosphate was similar in the three strains.This work was supported in part by a grant from the Ligue Nationale Française contre le Cancer (n^o 880258) to MS, and by a grant from the Fonds National Suisse de la Recherche Scientifique (No. 3.623-087) to CR.     

Gamete formation reflects the sexual pheromone hierarchy ofDictyostelium giganteum

Summary Sexual development inDictyostelium giganteum begins with the appearance of small, amoeboid gametes that fuse to produce mainly binucleate cells which differentiate into zygote giant cells. The data presented here show that the number of gametes produced by each strain (WS589>WS606>WS607>WS588) is directly related to its position in this hierarchy.This work was supported by a grant (A6807) from the Natural Sciences and Engineering Research Council of Canada.     

DNG1, a Dictyostelium homologue of tumor suppressor ING1 regulates differentiation of Dictyostelium cells

dng1 is a Dictyostelium homologue of the mammalian tumor suppressor ING gene. DNG1 protein localizes in the nucleus, and has a highly conserved PHD finger domain found in chromatin-remodeling proteins. Both dng1 disruption and overexpression impaired cell proliferation. In dng1-null cells, the progression of differentiation was delayed in a cell-density-dependent manner, and many tiny aggregates were formed. Exogenously applied cAMP pulses reversed the inhibitory effect caused by dng1 disruption on the aggregation during early development, but formation of tiny aggregates was not restored. dng1-overexpressing cells acquired the ability to undergo chemotaxis to cAMP earlier and exhibited enhanced differentiation. These phenotypes were found to be coupled with altered expressions of early genes such as cAMP receptor 1 (car1) and contact site A (csA). Furthermore, disordered histone modifications were demonstrated in dng1-null cells. These results suggest a regulatory role of dng1 in the transition of cells from growth to differentiation.Received 29 December 2004; received after revision 24 May 2005; accepted 26 May 2005     

Parameters affecting the maximum cell concentration ofTetrahymena

Summary In cultures with efficient aeration a maximum cell concentration (MCC) of 6×10⁵ cells/ml (defined medium) and 5.5×10⁶ cells/ml (broth) can be reached. By culturing within Millicells with excess supply of medium and efficient removal of waste products a physical limit for MCC of about 13×10⁶ cells/ml is reached.     

Analysis of the immune system with transgenic mice: T cell development

Transgenic mice carrying functionally rearranged T cell receptor genes have contributed significantly to our knowledge of T cell development and thymic positive and negative selection processes. In addition, TCR-transgenic mice have been used to investigate mutations affecting thymocyte development, likescid andlpr. Gene targeting by homologous recombination will allow to analyze more specifically the molecular mechanisms underlying thymic selection and peripheral tolerance.     

A new cell line (XTY) from a tumor ofXenopus laevis

A new cell line (XTY) was derived from a tumor of a femaleXenopus laevis. This cell line has been proliferating in standard amphibian culture medium for more than 4 years (470 generations) and has a doubling time of 75.5 h at 25°C. The cells aggregate into large groups, and their stellate morphology and the expression of desmin demonstrated by immunocytochemistry suggest that their origin is not epithelial.     

Gel-electrophoretic description of European populations ofTerellia virens (Loew) (Diptera,Tephritidae); implications for its use as an agent for the biological control ofCentaurea spp. (Asteraceae) in North America

Allozyme frequencies of 15 enzyme loci, 14 of which were polymorphic, were used to characterize sevenTerellia virens populations originating from three allopatrically distributedCentaurea species. The two populations whose origins were geographically furthest apart, from Israel (onC. iberica) and from Switzerland (onC. vallesiaca), showed relatively high values of genetic distance from the 5 populations sampled in Austria and Hungary (onC. maculosa) (Nei's D>0.07). The latter five displayed a high degree of genetic similarity. No diagnostic (fixed) allelic differences were observed between these three groups ofT. virens populations, but they could be well characterized by significant differences in allelic frequencies at 9 enzyme loci. Independently of this study, the populations from Switzerland (C. vallesiaca) and eastern Austria (C. maculosa) were selected as potential source populations for future introductions into North America for the biological control of introducedC. maculosa andC. diffusa. Based on the observed genetic differences and results from field experiments on the host specificity of these two potential source populations, it is argued that host specificity screening tests should be conducted separately for local (host plant) populations, as such populations might accept a different set of hosts. Biotype mismatch and the risk of spill-overs to native species could thus possibly be reduced.     

Arresting the mitotic oscillator and the control of cell proliferation: insights from a cascade model for cdc2 kinase activation

We consider a minimal cascade model previously proposed¹¹ for the mitotic oscillator driving the embryonic cell division cycle. The model is based on a bicyclic phosphorylation-dephosphorylation cascade involving cyclin and cdc2 kinase. By constructing stability diagrams showing domains of periodic behavior as a function of the maximum rates of the kinases and phosphatases involved in the two cycles of the cascade, we investigate the role of these converter enzymes in the oscillatory mechanism. Oscillations occur when the balance of kinase and phosphatase rates in each cycle is in a range bounded by two critical values. The results suggest ways to arrest the mitotic oscillator by altering the maximum rates of the converter enzymes. These results bear on the control of cell proliferation.     

Agglutinins from aquatic insects—tumor cell agglutination activity

Agglutinins were identified in whole body extracts of aquatic insects by means of murine tumor cell agglutination, using sarcoma 180 ascites, Ehrlich, and MM-46 cells. Screening revealed agglutinins in 5 of 10 of the larvae tested, and in 2 of 6 of the water-dwelling adult insects;Gerris paludum insularis andGyrinus japonicus. Only the agglutinin from adultG. paludum also agglutinated human erythrocytes. An ascites tumor was converted into a solid form in vivo after administration ofG. paludum agglutinin. The observation that these aquatic insect agglutinins preferentially agglutinate tumor cells has considerable implications in terms of anti-tumor effects such as inhibition of cell proliferation and metastasis.Deceased.     

Induction of cyst formation by low temperature in the dinoflagellateGonyaulax polyedra Stein: Dependence on circadian phase and requirement of light

Encystment, which at a temperature of 15°C is photoperiodically controlled inGonyaulax polyedra, can also be induced by a decrease of temperature, from 20 to 10 or 8°C in the absence of photoperiodic signals. The cyst-inducing capacity of the decrease in temperature depends on the circadian phase: in constant light, the maximum of sensitivity was found at the beginning of subjective night. In a light/dark cycle, however, cyst formation was reduced during dark phase, indicating that light is required for the process of encystment. A similar light dependence was seen in the effect of the physiologically occurring cyst inducer 5-methoxytryptamine, but not in the encystment response to the protonophores monensin and nigericin.     

The presence and function of melatonin and structurally related indoleamines in a dinoflagellate,and a hypothesis on the evolutionary significance of these tryptophan metabolites in unicellulars

The bioluminescent dinoflagellateGonyaulax polyedra contains various indoleamines, in particular, melatonin and 5-methoxytryptamine, as well as enzymes of their biosynthetic pathway. Melatonin exhibits a high-amplitude circadian rhythm characterized by a dramatic increase shortly after the onset of darkness. The maximum of melatonin is followed by a peak of 5-methoxytryptamine. These 5-methoxylated indoleamines seem to be involved in the mediation of the information darkness.G. polyedra shows a short-day response, which consists in the formation of asexual cysts. Light break experiments demonstrate the photoperiodic nature of this reaction. Cells become sensitive to short days only upon exposure to a lowered temperature (<16°C). Melatonin mimics the short-day effect, but only at decreased temperature. 5-Methoxytryptamine is even a better inducer of cyst formation, acting also at 20°C and in any lighting schedule, including LL. Cyst induction is associated with stimulation of bioluminescence and cytoplasmic acidification. A model on the intracellular pathway of photoperiodic information transduction assumes increased deacetylation of melatonin under cyst-inducing conditions, binding of 5-methoxytryptamine to the membrane of an acidic vacuole, proton transfer to the cytoplasm, and decreased intracellular pH as the stimulus for encystment. Melatonin shows the property of a scavenger of superoxide anions. This reaction, which is efficiently catalyzed by hemin, leads to the formation of a substituted kynuramine (AFMK). Destruction of melatonin by light-induced superoxide anions in the presence of cellular hemin may represent a property which, during evolution, has made this molecule suitable as an indicator of darkness. On the other hand, AFMK, which is formed under illumination, might have become a mediator of the information light. Photoperiodism inGonyaulax shows surprising parallels to that in mammals, but allows the analysis of this phenomenon at an entirely cellular level.     

Inhibition of virus multiplication and alteration of cyclic AMP level in cell cultures by flavonoids

Summary The inhibitory effect of four flavonoid compounds on virus multiplication and their influence on the intracellular cyclic AMP (cAMP) level were studied in cell cultures. Quercetin and quercitrin reduced the yields ofHuman (alpha) herpesvirus 1 (HSV-1) andSuid (alpha) herpesvirus 1 (pseudorabies virus), but hesperidin and rutin had no effect. Further, quercetin and quercitrin elevated the intracellular level of cAMP, whereas hesperidin and rutin did not alter the cAMP level. Both antiviral activity and cAMP-enhancing effect were dependent on the concentrations of the flavonoids, and these effects turned out to be parallel.This study suggests that a relation exists between the antiviral effect and the cAMP-enhancing activity of flavonoids.     

Amorphous calcium phosphate in the stylets produced by a marine worm (Nemertea)

Summary Electron microprobe analyses of the calcified stylets produced by the nemertean wormAmphiporus formidabilis reveal large calcium and phosphorus peaks. IR spectroscopy and X-ray diffraction indicate that the calcium and phosphorus in stylets constitute an amorphous calcium phosphate rather than a crystalline mineral phase.     

Replication pattern of double minutes derived from an insect cell line

Summary The DNA replication pattern of double minutes derived from an established cell line ofAedes albopictus is described. Although the vast majority of double minutes replicate semiconservatively once during the S phase, some double minutes appear to exhibit different pattern(s). Two theories are suggested as possible explanations of our findings.This work was supported by a grant from the Whitehall Foundation, Inc. awarded to A.B.M.     

Malarial parasites complete sporogony in axenic mosquitoes

Summary To obtain sporogonic stages of malaria free from microbial contaminants for in vitro studies,Anopheles stephensi were reared under sterile conditions using a mosquito cell line as larval food. The adult females, kept in sterile humidified containers and allowed to engorge on parasitemic hamsters, supported the sporogonic development of the rodent malarial parasitePlasmodium berghei. In 10 experiments, the proportion of infected mosquitoes varied from 0 to 92%, and the geometric mean number of oocysts per female mosquito from 2.5 to 58,6, with a range of 1 to 548. The average number of salivary gland sporozoites per infected mosquito was determined by direct sporozoite counts in the pooled homogenate of the thoraces of all female mosquitoes. In five experiments, it varied from 2.7×10³ to 9.0×10³. The sterile sporozoites, harvested on day 19 or 20 after the infective blood meal, were as infective for rodents as nonsterile ones.Supported in part by Public Health Service research grant AI 18345 from the National Institute of Allergy and Infectious Diseases, by a grant from the Agency of International Development DSPE-5542-G-SS-3042-00, and by a Charles and Johanna Busch award.