Involvement of Sib proteins in the regulation of cellular adhesion in Dictyostelium discoideum

Molecular mechanisms ensuring cellular adhesion have been studied in detail in Dictyostelium amoebae, but little is known about the regulation of cellular adhesion in these cells. Here, we show that cellular adhesion is regulated in Dictyostelium, notably by the concentration of a cellular secreted factor accumulating in the medium. This constitutes a quorum-sensing mechanism allowing coordinated regulation of cellular adhesion in a Dictyostelium population. In order to understand the mechanism underlying this regulation, we analyzed the expression of recently identified Dictyostelium adhesion molecules (Sib proteins) that present features also found in mammalian integrins. sibA and sibC are both expressed in vegetative Dictyostelium cells, but the expression of sibC is repressed strongly in conditions where cellular adhesion decreases. Analysis of sibA and sibC mutant cells further suggests that variations in the expression levels of sibC account largely for changes in cellular adhesion in response to environmental cues.     

集合种群动态对生境毁坏空间异质性的响应

首次将分形几何(Fractal geometry)与元胞自动机(Cellular automata)相结合,研究了破碎化生境中集合种群的空间分布格局动态,以及集合种群动态对生境毁坏空间异质性的响应。研究发现:(1)各个物种种群在生境中的分布具有很好的分形特征,物种的计盒维数(Box dimension)不仅可以很好地反映种群的空间分布结构,也能很好地反映种群动态。(2)如果将空间因素考虑进来的话,生境毁坏的灭绝债务(Time debt)将大于空间隐含模式所模拟的结果。(3)物种灭绝同时存在强物种灭绝和弱物种灭绝。并且只有在生境随机毁坏下,才与空间隐含的模拟结果比较接近,即强物种中将是最强物种率先灭绝。而在边缘毁坏这种比较集中成块的开发方式下,将是较强的物种灭绝。(4)边缘毁坏相对随机毁坏有利于物种,尤其是弱物种的长期续存。     

Towards a molecular understanding of human diseases using Dictyostelium discoideum

The social amoeba Dictyostelium discoideum is increasingly being used as a simple model for the investigation of problems that are relevant to human health. This article focuses on several recent examples of Dictyostelium-based biomedical research, including the analysis of immune-cell disease and chemotaxis, centrosomal abnormalities and lissencephaly, bacterial intracellular pathogenesis, and mechanisms of neuroprotective and anti-cancer drug action. The combination of cellular, genetic and molecular biology techniques that are available in Dictyostelium often makes the analysis of these problems more amenable to study in this system than in mammalian cell culture. Findings that have been made in these areas using Dictyostelium have driven research in mammalian systems and have established Dictyostelium as a powerful model for human-disease analysis.     

Total internal reflection fluorescence microscopy for single-molecule imaging in living cells

Marvelous background rejection in total internal reflection fluorescence microscopy (TIR-FM) has made it possible to visualize single-fluorophores in living cells. Cell signaling proteins including peptide hormones, membrane receptors, small G proteins, cytoplasmic kinases as well as small signaling compounds have been conjugated with single chemical fluorophore or tagged with green fluorescent proteins and visualized in living cells. In this review, the reasons why single-molecule analysis is essential for studies of intracellular protein systems such as cell signaling system are discussed, the instrumentation of TIR-FM for single-molecule imaging in living cells is explained, and how single molecule visualization has been used in cell biology is illustrated by way of two examples: signaling of epidermal growth factor in mammalian cells and chemotaxis of Dictyostelium amoeba along a cAMP gradient. Single-molecule analysis is an ideal method to quantify the parameters of reaction dynamics and kinetics of unitary processes within intracellular protein systems. Knowledge of these parameters is crucial for the understanding of the molecular mechanisms underlying intracellular events, thus single-molecule imaging in living cells will be one of the major technologies in cellular nanobiology.     

Antioxidant systems and anoxia tolerance in a freshwater turtle Trachemys scripta elegans

In the search for MBP phosphorylating activities in Dictyostelium discoideum, we have found a proteolysis-activated protein kinase. This activity which is distributed between the soluble and the particulate fractions of the cell, uses MBP and histone as substrate and has a molecular mass of 140 kDa as detected in an in situ' assay.This protein kinase has several features shared by the protein kinase C family, such as substrate specificity and sensitivity to proteolysis, but its molecular mass is much larger than that described for the known protein kinase C isoforms.To better characterize this activity we have studied its sensitivity to several protein kinase C inhibitors and activators. This protein kinase is activated neither by phorbol ester nor by phosphatidylserine or Ca²⁺. The activity is inhibited by staurosporine and PKC pseudosubstrate, but is not affected by the specific protein kinase C inhibitor bisindolylmaleimide.These data lead us to propose that proteolytically activated Dictyostelium protein kinase belongs to the recently described protein kinase C-related family.     

Myosin I contributes to the generation of resting cortical tension.

The amoeboid myosin I's are required for cellular cortical functions such as pseudopod formation and macropinocytosis, as demonstrated by the finding that Dictyostelium cells overexpressing or lacking one or more of these actin-based motors are defective in these processes. Defects in these processes are concomitant with changes in the actin-filled cortex of various Dictyostelium myosin I mutants. Given that the amoeboid myosin I's possess both actin- and membrane-binding domains, the mutant phenotypes could be due to alterations in the generation and/or regulation of cell cortical tension. This has been directly tested by analyzing mutant Dictyostelium that either lacks or overexpresses various myosin I's, using micropipette aspiration techniques. Dictyostelium cells lacking only one myosin I have normal levels of cortical tension. However, myosin I double mutants have significantly reduced (50%) cortical tension, and those that mildly overexpress an amoeboid myosin I exhibit increased cortical tension. Treatment of either type of mutant with the lectin concanavalin A (ConA) that cross-links surface receptors results in significant increases in cortical tension, suggesting that the contractile activity of these myosin I's is not controlled by this stimulus. These results demonstrate that myosin I's work cooperatively to contribute substantially to the generation of resting cortical tension that is required for efficient cell migration and macropinocytosis.     

Dynamic pattern processing with adaptive excitable media

We use an excitable medium for dynamic pattern processing. For this purpose the autowave structures arising from the light-sensitive Belouzov-Zhabotinsky reaction could serve as attractors. The dynamics is simulated as a non-linear network with local interaction in terms of cellular automata. With the help of a set of threshold electrodes the actual state is projected into a space of bytes. At the end of the adaptation procedure for the parameters we achieved the ability of this system for pattern recognition. Dynamic and statis pattern processing are compared with respect to information capacity and adaption time.     

空间显式模型模拟河流岸边带植被在水库运行作用下的演替

岸边带是水-陆之间的过渡和缓冲地带,是河流生态系统的重要组成部分.岸边带对拦截径流中的固体颗粒、吸收营养盐、减少入河污染负荷有重要作用.受河流水位季节性波动的影响,岸边带生态系统的变化非常剧烈,而当水库等水工建筑的运行剧烈改变河流的水文情势时,水库下游的岸边带生态系统将受到长期的累积性影响.因此,研究复式河道岸边带植被动态对于受损河流生态修复以及河流开发运行的生态环境影响规避具有重要意义.通过原位样方观测和室内水槽模拟试验,开发了岸边带植被演替模型,该模型耦合了全局基于连续性模式的水动力模块和局部基于元胞自动机模式的植被演替模块,并以漓江中游的一段复式河道为例,通过模拟水库运行前后长序列的水文情势变化和3种岸边带植物(刺果酸模、水蓼和益母草)的生长演替,分析了为满足旅游航道需求上游水库补水运行对下游岸边带植被的影响.     

盘基网柄菌——研究致病机制的宿主模型

盘基网柄菌作为致病菌宿主模型的研究主要有:筛选致病菌株及相应突变菌株毒性;鉴别对致病菌易感性和抗性的突变细胞宿主;宿主细胞的有效标记、已完成的基因组计划以及宿主细胞与致病菌间信号转导通路的相互作用;这些都表明盘基网柄菌是致病机制研究的理想宿主模型。     

Predicting the Distribution of Spiral Waves from Cell Properties in a Developmental-Path Model of Dictyostelium Pattern Formation

The slime mold Dictyostelium discoideum is one of the model systems of biological pattern formation. One of the most successful answers to the challenge of establishing a spiral wave pattern in a colony of homogeneously distributed D. discoideum cells has been the suggestion of a developmental path the cells follow (Lauzeral and coworkers). This is a well-defined change in properties each cell undergoes on a longer time scale than the typical dynamics of the cell. Here we show that this concept leads to an inhomogeneous and systematic spatial distribution of spiral waves, which can be predicted from the distribution of cells on the developmental path. We propose specific experiments for checking whether such systematics are also found in data and thus, indirectly, provide evidence of a developmental path.     

Pattern recall in networks of chaotic neurons

This research investigates the potential utility of chaotic dynamics in neural information processing. A novel chaotic spiking neural network model is presented which is composed of non-linear dynamic state (NDS) neurons. The activity of each NDS neuron is driven by a set of non-linear equations coupled with a threshold based spike output mechanism. If time-delayed self-connections are enabled then the network stabilises to a periodic pattern of activation. Previous publications of this work have demonstrated that the chaotic dynamics which drive the network activity ensure that an extremely large number of such periodic patterns can be generated by this network. This paper presents a major extension to this model which enables the network to recall a pattern of activity from a selection of previously stabilised patterns.     

Naringenin is a novel inhibitor of Dictyostelium cell proliferation and cell migration

Naringenin is a flavanone compound that alters critical cellular processes such as cell multiplication, glucose uptake, and mitochondrial activity. In this study, we used the social amoeba, Dictyostelium discoideum, as a model system for examining the cellular processes and signaling pathways affected by naringenin. We found that naringenin inhibited Dictyostelium cell division in a dose-dependent manner (IC(50) approximately 20 microM). Assays of Dictyostelium chemotaxis and multicellular development revealed that naringenin possesses a previously unrecognized ability to suppress amoeboid cell motility. We also found that naringenin, which is known to inhibit phosphatidylinositol 3-kinase activity, had no apparent effect on phosphatidylinositol 3,4,5-trisphosphate synthesis in live Dictyostelium cells; suggesting that this compound suppresses cell growth and migration via alternative signaling pathways. In another context, the discoveries described here highlight the value of using the Dictyostelium model system for identifying and characterizing the mechanisms by which naringenin, and related compounds, exert their effects on eukaryotic cells.     

Temporal and spatial expression of ammonium transporter genes during growth and development of Dictyostelium discoideum

Ammonia is an important signaling molecule involved in the regulation of development in Dictyostelium. During aggregation, ammonia gradients are established, and the ammonia concentration in the immediate environment or within a particular cell throughout development may vary. This is due to the rate of cellular ammonia production, its rate of loss by evaporation to the atmosphere or by diffusion into the substratum, and perhaps to cellular transport by ammonium transporters (AMTs). Recent efforts in genome and cDNA sequencing have identified three ammonium transporters in Dictyostelium. In addition to physically altering the levels of ammonia within cells, AMTs also may play a role in ammonia signaling. As an initial step in identifying such a function, the temporal and spatial expression of the three amt genes is examined. RT-PCR demonstrates that each of the three amt mRNAs is present and relatively constant throughout growth and development. The spatial expression of these three amt genes is examined during multiple stages of Dictyostelium development using in situ hybridization. A distinct and dynamic pattern of expression is seen for the three genes. In general, amtA is expressed heavily in pre-stalk cells in a dynamic way, while amtB and amtC are expressed in pre-spore regions consistently throughout development. AmtC also is expressed in the most anterior tip of fingers and slugs, corresponding to cells that mediate ammonia's effect on the choice between slug migration and culmination. Indeed, amtC null cells have a slugger phenotype, suggesting AmtC functions in the signaling pathway underlying the mechanics of this choice.     

Spiking behavior and epileptiform oscillations in a discrete model of cortical neural networks

Summary We investigate the phenomenon of epileptiform activity using a discrete model of cortical neural networks. Our model is reduced to the elementary features of neurons and assumes simplified dynamics of action potentials and postsynaptic potentials. The discrete model provides a comparably high simulation speed which allows the rendering of phase diagrams and simulations of large neural networks in reasonable time. Further the reduction to the basic features of neurons provides insight into the essentials of a possible mechanism of epilepsy. Our computer simulations suggest that the detailed dynamics of postsynaptic and action potentials are not indispensable for obtaining epileptiform behavior on the system level. The simulation results of autonomously evolving networks exhibit a regime in which the network dynamics spontaneously switch between fluctuating and oscillating behavior and produce isolated network spikes without external stimulation. Inhibitory neurons have been found to play an important part in the synchronization of neural firing: an increased number of synapses established by inhibitory neurons onto other neurons induces a transition to the spiking regime. A decreased frequency accompanying the hypersynchronous population activity has only occurred with slow inhibitory postsynaptic potentials.     

A demonstration of pattern formation without positional information in Dictyostelium

Although positional information, conveyed by morphogen gradients, is a widely accepted way of forming patterns during development, an alternative method is conceivable, based on the intermingled differentiation of cells with different fates, followed by their sorting into discrete pattern elements. It has been proposed that Dictyostelium prestalk and prespore cells behave in this way at the mound stage of development. However, it has been difficult to conclusively demonstrate that they initially differentiate intermingled, because rapid cell movement within the mound makes it impossible to be sure where prestalk and prespore cells originate. We have taken a novel approach to address this problem by blocking cell movement at different stages in development, using the actin-depolymerizing drug, latrunculin-A. Prestalk and prespore cells differentiate with essentially normal efficiency and timing in such paralyzed structures. When movement is blocked sufficiently early, the major cell types all subsequently differentiate at scattered positions throughout the aggregate, and even in the streams leading into it. Our work strongly supports the idea that the prestalk/prespore pattern in Dictyostelium forms without positional information and demonstrate that latrunculin-A may provide a useful tool for the investigation of patterning in other organisms.     

Inositol 1,4,5-trisphosphate induces cyclic GMP formation in Dictyostelium discoideum

Chemotactic signalling in the cellular slime mould Dictyostelium discoideum employs signalling molecules such as folate and cyclic AMP. These bind to specific cell surface receptors and rapidly trigger internal responses that induce chemotactic movement of the amoebae. Previous studies have shown that actin is polymerised within 3-5 sec of cyclic AMP or folate binding and that a peak of cyclic GMP is formed within 9-12 sec. Release of Ca2+ from intracellular stores has been implicated as a secondary messenger. Here we present evidence that D-myo-inositol 1,4,5-trisphosphate, when added to permeabilized amoebae of Dictyostelium, can mimic the action of chemoattractants on normal intact amoebae in inducing cyclic GMP formation. Our data suggest that IP3, which is known to act as an intermediary messenger between cell surface hormone receptors and release of Ca2+ from internal stores in mammalian cells, functions in a similar capacity during chemotaxis of this primitive eukaryote.     

Sexual Development of Cellular Slime Molds

Macrocyst formation represents sexual development of cellular slime molds and begins with fusion between cells of compatible mating types. Homothallic and heterothallic strains as well as bisexual and asexual ones have been described. Macrocyst development requires certain environmental conditions such as darkness and excessive humidity. Sexual cell fusion has been analyzed at a molecular level in Dictyostelium discoideum , and several cell surface proteins related to it have been identified. Some of them are common to both mating types, while others are specific to one or other type. The involvement of cell-surface carbohydrates has also been suggested, though direct evidence for this is still lacking. Macrocyst formation is regulated by diffusible, pheromone like substances. Genetic studies on sexual development are scarce, probably because no suitable mutants have been available. However, several asexual mutants, as well as antibody and nucleotide probes, have recently been obtained, so mechanisms of sexual cell fusion may be understood in the near future. Considering the unique phylogenical position of cellular slime molds, analysis of sexual development in these organisms should contribute to the understanding of the mechanism and evolution of sexual reproduction systems.