Pattern formation: fruiting body morphogenesis in Myxococcus xanthus

When Myxococcus xanthus cells are exposed to starvation, they respond with dramatic behavioral changes. The expansive swarming behavior stops and the cells begin to aggregate into multicellular fruiting bodies. The cell-surface-associated C-signal has been identified as the signal that induces aggregation. Recently, several of the components in the C-signal transduction pathway have been identified and behavioral analyses are beginning to reveal how the C-signal modulates cell behavior. Together, these findings provide a framework for understanding how a cell-surface-associated morphogen induces pattern formation.     

Cell polarity, intercellular signalling and morphogenetic cell movements in Myxococcus xanthus

In Myxococcus xanthus morphogenetic cell movements constitute the basis for the formation of spreading vegetative colonies and fruiting bodies in starving cells. M. xanthus cells move by gliding and gliding motility depends on two polarly localized engines, type IV pili pull cells forward, and slime extruding nozzle-like structures appear to push cells forward. The motility behaviour of cells provides evidence that the two engines are localized to opposite poles and that they undergo polarity switching. Several proteins involved in regulating polarity switching have been identified. The cell surface-associated C-signal induces the directed movement of cells into nascent fruiting bodies. Recently, the molecular nature of the C-signal molecule was elucidated and the motility parameters regulated by the C-signal were identified. From the effect of the C-signal on cell behaviour it appears that the C-signal inhibits polarity switching of the two motility engines. This establishes a connection between cell polarity, signalling by an intercellular signal and morphogenetic cell movements during fruiting body formation.     

Making waves: pattern formation by a cell-surface-associated signal

Starving Myxococcus xanthus cells organize into two strikingly different spatio-temporal patterns, either rippling or aggregation of cells into fruiting bodies. Formation of both patterns depends on a cell-surface-associated, non-diffusible signal, the C-signal. A key motility parameter modulated by the C-signal during pattern formation is the frequency at which cells reverse their gliding direction, with low and high levels of C-signalling causing an increase and a decrease in the reversal frequency, respectively. Recently, a simple yet elegant mathematical model was proposed to explain the mechanism underlying the non-linear dependence of the reversal frequency on C-signalling levels. The mathematical solution hinges on the introduction of a negative feedback loop into the biochemical circuit that regulates the reversal frequency. This system displays an oscillatory behaviour in which the oscillation frequency depends in a non-monotonic manner on the level of C-signalling. Thus, the biochemical oscillator recapitulates the effect of the C-signal on the reversal frequency. The challenge for biologists now is to test the mathematical model experimentally.     

Cell behavior and cell-cell communication during fruiting body morphogenesis in Myxococcus xanthus

Formation of spatial patterns of cells from a mass of initially identical cells is a recurring theme in developmental biology. The dynamics that direct pattern formation in biological systems often involve morphogenetic cell movements. An example is fruiting body formation in the gliding bacterium Myxococcus xanthus in which an unstructured population of identical cells rearranges into an asymmetric, stable pattern of multicellular fruiting bodies in response to starvation. Fruiting body formation depends on changes in organized cell movements from swarming to aggregation. The aggregation process is induced and orchestrated by the cell-surface associated 17 kDa C-signal protein. C-signal transmission depends on direct contact between cells. Evidence suggests that C-signal transmission is geometrically constrained to cell ends and that productive C-signal transmission only occurs when cells engage in end-to-end contacts. Here, we review recent progress in the understanding of the pattern formation process that leads to fruiting body formation. Gliding motility in M. xanthus involves two polarly localized gliding machines, the S-machine depends on type IV pili and the A-machine seems to involve a slime extrusion mechanism. Using time-lapse video microscopy the gliding motility parameters controlled by the C-signal have been identified. The C-signal induces cells to move with increased gliding speeds, in longer gliding intervals and with decreased stop and reversal frequencies. The combined effect of the C-signal dependent changes in gliding motility behaviour is an increase in the net-distance travelled by a cell per minute. The identification of the motility parameters controlled by the C-signal in combination with the contact-dependent C-signal transmission mechanism have allowed the generation of a qualitative model for C-signal induced aggregation. In this model, the directive properties of the C-signal are a direct consequence of the contact-dependent signal-transmission mechanism, which is a local event involving direct contact between cells that results in a global organization of cells. This pattern formation process does not depend on a diffusible substance. Rather it depends on a cell-surface associated signal to direct the cells appropriately.     

C-signal: a cell surface-associated morphogen that induces and co-ordinates multicellular fruiting body morphogenesis and sporulation in Myxococcus xanthus

In Myxococcus xanthus, morphogenesis of multicellular fruiting bodies and sporulation are co-ordinated temporally and spatially. csgA mutants fail to synthesize the cell surface-associated C-signal and are unable to aggregate and sporulate. We report that csgA encodes two proteins, a 25 kDa species corresponding to full-length CsgA protein and a 17 kDa species similar in size to C-factor protein, which has been shown previously to have C-signal activity. By systematically varying the accumulation of the csgA proteins, we show that overproduction of the csgA proteins results in premature aggregation and sporulation, uncoupling of the two events and the formation of small fruiting bodies, whereas reduced synthesis of the csgA proteins causes delayed aggregation, reduced sporulation and the formation of large fruiting bodies. These results show that C-signal induces aggregation as well as sporulation, and that an ordered increase in the level of C-signalling during development is essential for the spatial co-ordination of these events. The results support a quantitative model, in which aggregation and sporulation are induced at distinct threshold levels of C-signalling. In this model, the two events are temporally co-ordinated by the regulated increase in C-signalling levels during development. The contact-dependent C-signal transmission mechanism allows the spatial co-ordination of aggregation and sporulation by coupling cell position and signalling levels.     

The act operon controls the level and time of C-signal production for Myxococcus xanthus development

The C-signal is a morphogen that controls the assembly of fruiting bodies and the differentiation of myxospores. Production of this signal, which is encoded by the csgA gene, is regulated by the act operon of four genes that are co-transcribed from the same start site. The act A and act B genes regulate the maximum level of the C-signal, which never rises above one-quarter of the maximum wild-type level of CsgA protein in null mutants of either gene. The act A and act B mutants have the same developmental phenotype: both aggregate, neither sporulates, both prolong rippling. By sequence homology, act A encodes a response regulator, and act B encodes a sigma-54 activator protein of the NTRC class. The similar phenotypes of act A and act B deletion mutants suggest that the two gene products are part of the same signal transduction pathway. That pathway responds to C-signal and also regulates the production of CsgA protein, thus creating a positive feedback loop. The act C and act D genes regulate the time pattern of CsgA production, while achieving the same maximum level. An act C null mutant raises CsgA production 15 h earlier than the wild type, whereas an act D null mutant does so 6 h later than wild type. The loop explains how the C-signal rises continuously from early development to a peak at the time of sporulation, and the act genes govern the time course of that rise.     

Trypanosome hydrolases and the blood-brain barrier

African trypanosomes cross the blood-brain barrier, but how they do so remains an area of speculation. We propose that proteases, such as the trypanopains and oligopeptidases that are released by trypanosomes, could mediate in this process. The trypanosomes also possess cell-surface-associated acid phosphatases that could play a role in invasion similar to that in advancing cancer cells. Such enzymes, perhaps acting in concert, have the potential to cause tissue degradation and ease the passage of the trypanosomes through various tissues in the host, including the blood-brain barrier.     

Mutational analysis of the Myxococcus xanthus Omicron4499 promoter region reveals shared and unique properties in comparison with other C-signal-dependent promoters

The bacterium Myxococcus xanthus undergoes multicellular development during times of nutritional stress and uses extracellular signals to coordinate cell behavior. C-signal affects gene expression late in development, including that of Omega4499, an operon identified by insertion of Tn5 lac into the M. xanthus chromosome. The Omega4499 promoter region has several sequences in common with those found previously to be important for expression of other C-signal-dependent promoters. To determine if these sequences are important for Omega4499 promoter activity, the effects of mutations on expression of a downstream reporter gene were tested in M. xanthus. Although the promoter resembles those recognized by Escherichia coli sigma(54), mutational analysis implied that a sigma(70)-type sigma factor likely recognizes the promoter. A 7-bp sequence known as a C box and a 5-bp element located 6 bp upstream of the C box have been shown to be important for expression of other C-signal-dependent promoters. The Omega4499 promoter region has C boxes centered at -33 and -55 bp, with 5-bp elements located 7 and 8 bp upstream, respectively. A multiple-base-pair mutation in any of these sequences reduced Omega4499 promoter activity more than twofold. Single base-pair mutations in the C box centered at -33 bp yielded a different pattern of effects on expression than similar mutations in other C boxes, indicating that each functions somewhat differently. An element from about -81 to -77 bp exerted a twofold positive effect on expression but did not appear to be responsible for the C-signal dependence of the Omega4499 promoter. Mutations in sigD and sigE, which are genes that encode sigma factors, reduced expression from the Omega4499 promoter. The results provide further insight into the regulation of C-signal-dependent genes, demonstrating both shared and unique properties among the promoter regions so far examined.     

An individual based model of rippling movement in a myxobacteria population

Migrating cells of Myxococcus xanthus (MX) in the early stages of starvation-induced development exhibit elaborate patterns of propagating waves. These so-called rippling patterns are formed by two sets of waves travelling in opposite directions. It has been experimentally shown that formation of these waves is mediated by cell-cell contact signalling (C-signalling). Here, we develop an individual-based model to study the formation of rippling patterns in MX populations. Following the work of Igoshin et al. (Proc. Natl. Acad. Sci. 98 (2001) 14913) we consider each moving cell to have an internal clock which controls its turning behaviour and sensitivity to C-signal. Specifically, we examine the effects of changing: C-signal strength, sensitivity/refractoriness, cell density, and noise upon the formation and structure of the rippling patterns. We also consider three modified models that have no explicit refractory period and examine their ability to produce rippling patterns.     

Recent Developments in Human Behavioral Genetics: Past Accomplishments and Future Directions

The field of behavioral genetics has enormous potential to uncover both genetic and environmental influences on normal and deviant behavior. Behavioral-genetic methods are based on a solid foundation of theories and methods that successfully have delineated components of complex traits in plants and animals. New resources are now available to dissect the genetic component of these complex traits. As specific genes are identified, we can begin to explore how these interact with environmental factors in development. How we interpret such findings, how we ask new questions, how we celebrate the knowledge, and how we use or misuse this knowledge are all important considerations. These issues are pervasive in all areas of human research, and they are especially salient in human behavioral genetics.     

Premotor neurons in the feeding system of Aplysia californica

Central pattern generator (CPG) circuits control cyclic motor output underlying rhythmic behaviors. Although there have been extensive behavioral and cellular studies of food-induced feeding arousal as well as satiation in Aplysia, very little is known about the neuronal circuits controlling rhythmic consummatory feeding behavior. However, recent studies have identified premotor neurons that initiate and maintain buccal motor programs underlying ingestion and egestion in Aplysia. Other newly identified neurons receive synaptic input from feeding CPGs and in turn synapse with and control the output of buccal motor neurons. Some of these neurons and their effects within the buccal system are modulated by endogenous neuropeptides. With this information we can begin to understand how neuronal networks control buccal motor output and how their activity is modulated to produce flexibility in observed feeding behavior.     

The staphylococcal transferrin receptor: a glycolytic enzyme with novel functions

To obtain iron from the host for growth, staphylococci have evolved sophisticated iron-scavenging systems including siderophores and a cell surface receptor for transferrin, the mammalian iron-transporting glycoprotein. The staphylococcal transferrin receptor has been identified as a member of a newly emerging family of multifunctional, cell-surface-associated glyceraldehyde-3-phosphate dehydrogenases, which not only retain their glycolytic enzyme activities but also bind diverse human serum proteins and possess NAD-ribosylating activity. These multiple functions suggest a potential contribution to virulence far beyond iron acquisition.     

Sex differences in social interaction behavior following social defeat stress in the monogamous California mouse (Peromyscus californicus)

Stressful life experiences are known to be a precipitating factor for many mental disorders. The social defeat model induces behavioral responses in rodents (e.g. reduced social interaction) that are similar to behavioral patterns associated with mood disorders. The model has contributed to the discovery of novel mechanisms regulating behavioral responses to stress, but its utility has been largely limited to males. This is disadvantageous because most mood disorders have a higher incidence in women versus men. Male and female California mice (Peromyscus californicus) aggressively defend territories, which allowed us to observe the effects of social defeat in both sexes. In two experiments, mice were exposed to three social defeat or control episodes. Mice were then behaviorally phenotyped, and indirect markers of brain activity and corticosterone responses to a novel social stimulus were assessed. Sex differences in behavioral responses to social stress were long lasting (4 wks). Social defeat reduced social interaction responses in females but not males. In females, social defeat induced an increase in the number of phosphorylated CREB positive cells in the nucleus accumbens shell after exposure to a novel social stimulus. This effect of defeat was not observed in males. The effects of defeat in females were limited to social contexts, as there were no differences in exploratory behavior in the open field or light-dark box test. These data suggest that California mice could be a useful model for studying sex differences in behavioral responses to stress, particularly in neurobiological mechanisms that are involved with the regulation of social behavior.