Unconventional signalling in tunicates

The zooids in colonial tunicates do not appear to be directly interconnected by nerves, but this has not prevented the evolution of coordinated behaviour in several groups. In Botryllus and other colonial styelid asci‐dians the endothelium lining the blood vessels is excitable and transmits action potentials from cell to cell via gap junctions. These signals mediate protective contractions of the zooids and synchronize contractions of the vascular ampullae. In didemnid ascidians such as Diplosoma a network of myocytes in the tunic serves to transmit excitation and to cause contractions of the cloacal apertures. Individual zooids of Pyrosoma protect themselves by closing their siphons and arresting their branchial cilia when stimulated. At the same time a flash of light is emitted. Neighbouring zooids sense the flash with their photoreceptors and respond in turn with protective responses and light emission. Protective responses thus spread by photic signalling and propagate from zooid to zooid through the colony in a saltatory manner. In chains of Salpafusifortnis, changes in the direction and/or speed of swimming are transmitted from zooid to zooid via adhesion plaques. When a zooid is stimulated, its body‐wall epithelium conducts action potentials to the plaque connecting it to the next zooid, exciting receptor neurons in that zooid. These receptors have sensory processes that bridge the gap between the two zooids. The sensory neurons so excited in the second zooid conduct impulses to the brain where they alter the motor output pattern, and at the same time generate epithelial action potentials that travel to the next zooid in line, where the same thing happens.

It is not clear why these unconventional signalling methods have evolved but the tunic may be an inhospitable environment for nerves, making conventional nervous links impossible.     

From individual Wnt pathways towards a Wnt signalling network

Wnt proteins play important roles during vertebrate and invertebrate development. They obviously have the ability to activate different intracellular signalling pathways. Based on the characteristic intracellular mediators used, these are commonly described as the Wnt/beta-catenin, the Wnt/calcium and the Wnt/Jun N-terminal kinase pathways (also called planar cell polarity pathway). In the past, these different signalling events were mainly described as individual and independent signalling branches. Here, we discuss the possibility that Wnt proteins activate a complex intracellular signalling network rather than individual pathways and suggest a graph representation of this network. Furthermore, we discuss different ways of how to predict the specific outcome of an activation of this network in a particular cell type, which will require the use of mathematical models. We point out that the use of deterministic approaches via the application of differential equations is suitable to model only small aspects of the whole network and that more qualitative approaches are possibly a suitable starting point for the prediction of the global behaviour of such large protein interaction networks.     

Representational signalling in birds

Some animals give specific calls when they discover food or detect a particular type of predator. Companions respond with food-searching behaviour or by adopting appropriate escape responses. These signals thus seem to denote objects in the environment, but this specific mechanism has only been demonstrated for monkey alarm calls. We manipulated whether fowl (Gallus gallus) had recently found a small quantity of preferred food and then tested for a specific interaction between this event and their subsequent response to playback of food calls. In one treatment, food calls thus potentially provided information about the immediate environment, while in the other the putative message was redundant with individual experience. Food calls evoked substrate searching, but only if the hens had not recently discovered food. An identical manipulation had no effect on responses to an acoustically matched control call. These results show that chicken food calls are representational signals: they stimulate retrieval of information about a class of external events. This is the first such demonstration for any non-primate species. Representational signalling is hence more taxonomically widespread than has previously been thought, suggesting that it may be the product of common social factors, rather than an attribute of a particular phylogenetic lineage.     

From components to regulatory motifs in signalling networks.

The developments in biochemistry and molecular biology over the past 30 years have produced an impressive parts list of cellular components. It has become increasingly clear that we need to understand how components come together to form systems. One area where this approach has been growing is cell signalling research. Here, instead of focusing on individual or small groups of signalling proteins, researchers are now using a more holistic perspective. This approach attempts to view how many components are working together in concert to process information and to orchestrate cellular phenotypic changes. Additionally, the advancements in experimental techniques to measure and visualize many cellular components at once gradually grow in diversity and accuracy. The multivariate data, produced by experiments, introduce new and exciting challenges for computational biologists, who develop models of cellular systems made up of interacting cellular components. The integration of high-throughput experimental results and information from legacy literature is expected to produce computational models that would rapidly enhance our understanding of the detail workings of mammalian cells.     

Computational models of signalling networks for non-linear control

Artificial signalling networks (ASNs) are a computational approach inspired by the signalling processes inside cells that decode outside environmental information. Using evolutionary algorithms to induce complex behaviours, we show how chaotic dynamics in a conservative dynamical system can be controlled. Such dynamics are of particular interest as they mimic the inherent complexity of non-linear physical systems in the real world. Considering the main biological interpretations of cellular signalling, in which complex behaviours and robust cellular responses emerge from the interaction of multiple pathways, we introduce two ASN representations: a stand-alone ASN and a coupled ASN. In particular we note how sophisticated cellular communication mechanisms can lead to effective controllers, where complicated problems can be divided into smaller and independent tasks.     

Identification of potential targets in biological signalling systems through network perturbation analysis

The network-based representation and analysis of biological systems contributes to a greater understanding of their structures and functions at different levels of complexity. These techniques can also be used to identify potential novel therapeutic targets based on the characterisation of vulnerable or highly influential network components. There is a need to investigate methods for estimating the impact of molecular perturbations. The prediction of high-impact or critical targets can aid in the identification of novel strategies for controlling the level of activation of specific, therapeutically relevant genes or proteins. Here, we report a new computational strategy for the analysis of the vulnerability of cellular signalling networks based on the quantitative assessment of the impact of large-scale, dynamic perturbations. To show the usefulness of this methodology, two complex signalling networks were analysed: the caspase-3 and the adenosine-regulated calcium signalling systems. This allowed us to estimate and rank the perturbation impact of the components defining these networks. Testable hypotheses about how these targets could modify the dynamic operation of the systems are provided. In the case of the caspase-3 system, the predictions and rankings were in line with results obtained from previous experimental validations of computational predictions generated by a relatively more computationally complex technique. In the case of the adenosine-regulated calcium system, we offer new testable predictions on the potential effect of different targets on the control of calcium flux. Unlike previous methods, the proposed approach provides perturbation-specific scores for each network component. The proposed perturbation assessment methodology may be applied to other systems to gain a deeper understanding of their dynamic operation and to assist the discovery of new therapeutic targets and strategies.     

Signal verification can promote reliable signalling

The central question in communication theory is whether communication is reliable, and if so, which mechanisms select for reliability. The primary approach in the past has been to attribute reliability to strategic costs associated with signalling as predicted by the handicap principle. Yet, reliability can arise through other mechanisms, such as signal verification; but the theoretical understanding of such mechanisms has received relatively little attention. Here, we model whether verification can lead to reliability in repeated interactions that typically characterize mutualisms. Specifically, we model whether fruit consumers that discriminate among poor- and good-quality fruits within a population can select for reliable fruit signals. In our model, plants either signal or they do not; costs associated with signalling are fixed and independent of plant quality. We find parameter combinations where discriminating fruit consumers can select for signal reliability by abandoning unprofitable plants more quickly. This self-serving behaviour imposes costs upon plants as a by-product, rendering it unprofitable for unrewarding plants to signal. Thus, strategic costs to signalling are not a prerequisite for reliable communication. We expect verification to more generally explain signal reliability in repeated consumer–resource interactions that typify mutualisms but also in antagonistic interactions such as mimicry and aposematism.     

Calcium signalling in early embryos

The onset of development in most species studied is triggered by one of the largest and longest calcium transients known to us. It is the most studied and best understood aspect of the calcium signals that accompany and control development. Its properties and mechanisms demonstrate what embryos are capable of and thus how the less-understood calcium signals later in development may be generated. The downstream targets of the fertilization calcium signal have also been identified, providing some pointers to the probable targets of calcium signals further on in the process of development.In one species or another, the fertilization calcium signal involves all the known calcium-releasing second messengers and many of the known calcium-signalling mechanisms. These calcium signals also usually take the form of a propagating calcium wave or waves. Fertilization causes the cell cycle to resume, and therefore fertilization signals are cell-cycle signals. In some early embryonic cell cycles, calcium signals also control the progress through each cell cycle, controlling mitosis.Studies of these early embryonic calcium-signalling mechanisms provide a background to the calcium-signalling events discussed in the articles in this issue.     

The signalling mucin Msb2 regulates surface sensing and host penetration via BMP1 MAP kinase signalling in Botrytis cinerea

Botrytis cinerea is a necrotrophic fungus that infects a wide range of fruit, vegetable and flower crops. Penetration of the host cuticle occurs via infection structures that are formed in response to appropriate plant surface signals. The differentiation of these structures requires a highly conserved mitogen‐activated protein (MAP) kinase cascade including the MAP kinase BMP1. In yeast and several plant‐pathogenic fungi, the signalling mucin Msb2 has been shown to be involved in surface recognition and MAP kinase activation. In this study, a B. cinerea msb2 mutant was generated and characterized. The mutant showed normal growth, sporulation, sclerotia formation and stress resistance. In the absence of nutrients, abnormal germination with multiple germ tubes was observed. In the presence of sugars, normal germination occurred, but msb2 germlings were almost unable to form appressoria or infection cushions on hard surfaces. Nevertheless, the msb2 mutant showed only a moderate delay in lesion formation on different host plants, and formed expanding lesions similar to the wild‐type. Although the wild‐type showed increasing BMP1 phosphorylation during the first hours of germination on hard surfaces, the phosphorylation levels in the msb2 mutant were strongly reduced. Several genes encoding secreted proteins were found to be co‐regulated by BMP1 and Msb2 during germination. Taken together, B. cinerea Msb2 is likely to represent a hard surface sensor of germlings and hyphae that triggers infection structure formation via the activation of the BMP1 MAP kinase pathway.     

Sustained calcium signalling and caspase-3 activation involve NMDA receptors in thymocytes in contact with dendritic cells

L-glutamate, the major excitatory neurotransmitter, also has a role in non-neuronal tissues and modulates immune responses. Whether NMDA receptor (NMDAR) signalling is involved in T-cell development is unknown. In this study, we show that mouse thymocytes expressed an array of glutamate receptors, including NMDARs subunits. Sustained calcium (Ca²⁺) signals and caspase-3 activation in thymocytes were induced by interaction with antigen-pulsed dendritic cells (DCs) and were inhibited by NMDAR antagonists MK801 and memantine. NMDARs were transiently activated, triggered the sustained Ca²⁺ signal and were corecruited with the PDZ-domain adaptor postsynaptic density (PSD)-95 to thymocyte-DC contact zones. Although T-cell receptor (TCR) activation was sufficient for relocalization of NMDAR and PSD-95 at the contact zone, NMDAR could be activated only in a synaptic context. In these T-DC contacts, thymocyte activation occurred in the absence of exogenous glutamate, indicating that DCs could be a physiological source of glutamate. DCs expressed glutamate, glutamate-specific vesicular glutamate transporters and were capable of fast glutamate release through a Ca²⁺-dependent mechanism. We suggest that glutamate released by DCs could elicit focal responses through NMDAR-signalling in T cells undergoing apoptosis. Thus, synapses between T and DCs could provide a functional platform for coupling TCR activation and NMDAR signalling, which might reflect on T-cell development and modulation of the immune response.