Rachel McMullan

The Gα12-RGS RhoGEF-RhoA signalling pathway regulates neurotransmitter release in C. elegans

In Caenorhabditis elegans adults, the single Rho GTPase orthologue, RHO‐1, stimulates neurotransmitter release at synapses. We show that one of the pathways acting upstream of RHO‐1 in acetylcholine (ACh)‐releasing motor neurons depends on Gα12 (GPA‐12), which acts via the single C. elegans RGS RhoGEF (RHGF‐1). Activated GPA‐12 has the same effect as activated RHO‐1, inducing the accumulation of diacylglycerol and the neuromodulator UNC‐13 at release sites, and increased ACh release. We showed previously that RHO‐1 stimulates ACh release by two separate pathways—one that requires UNC‐13 and a second that does not. We show here that a non‐DAG‐binding‐UNC‐13 mutant that partially blocks increased ACh release by activated RHO‐1 completely blocks increased ACh release by activated GPA‐12. Thus, the upstream GPA‐12/RHGF‐1 pathway stimulates only a subset of RHO‐1 downstream effectors, suggesting that either the RHO‐1 effectors require different levels of activated RHO‐1 for activation or there are two distinct pools of RHO‐1 within C. elegans neurons.

Serotonergic Chemosensory Neurons Modify the C. elegans Immune Response by Regulating G-Protein Signaling in Epithelial Cells

The nervous and immune systems influence each other, allowing animals to rapidly protect themselves from changes in their internal and external environment. However, the complex nature of these systems in mammals makes it difficult to determine how neuronal signaling influences the immune response. Here we show that serotonin, synthesized in Caenorhabditis elegans chemosensory neurons, modulates the immune response. Serotonin released from these cells acts, directly or indirectly, to regulate G-protein signaling in epithelial cells. Signaling in these cells is required for the immune response to infection by the natural pathogen Microbacterium nematophilum. Here we show that serotonin signaling suppresses the innate immune response and limits the rate of pathogen clearance. We show that C. elegans uses classical neurotransmitters to alter the immune response. Serotonin released from sensory neurons may function to modify the immune system in response to changes in the animals external environment such as the availability, or quality, of food.

The RHO-1 RhoGTPase Modulates Fertility and Multiple Behaviors in Adult C. elegans

The Rho family of small GTPases are essential during early embryonic development making it difficult to study their functions in adult animals. Using inducible transgenes expressing either a constitutively active version of the single C. elegans Rho ortholog, RHO-1, or an inhibitor of endogenous Rho (C3 transferase), we demonstrate multiple defects caused by altering Rho signaling in adult C. elegans. Changes in RHO-1 signaling in cholinergic neurons affected locomotion, pharyngeal pumping and fecundity. Changes in RHO-1 signaling outside the cholinergic neurons resulted in defective defecation, ovulation, and changes in C. elegans body morphology. Finally both increased and decreased RHO-1 signaling in adults resulted in death within hours. The multiple post-developmental roles for Rho in C. elegans demonstrate that RhoA signaling pathways continue to be used post-developmentally and the resulting phenotypes provide an opportunity to further study post-developmental Rho signaling pathways using genetic screens.

Neuronal and non-neuronal signals regulate Caernorhabditis elegans avoidance of contaminated food

One way in which animals minimize the risk of infection is to reduce their contact with contaminated food. Here, we establish a model of pathogen-contaminated food avoidance using the nematode worm Caernorhabditis elegans. We find that avoidance of pathogen-contaminated food protects C. elegans from the deleterious effects of infection and, using genetic approaches, demonstrate that multiple sensory neurons are required for this avoidance behaviour. In addition, our results reveal that the avoidance of contaminated food requires bacterial adherence to non-neuronal cells in the tail of C. elegans that are also required for the cellular immune response. Previous studies in C. elegans have contributed significantly to our understanding of molecular and cellular basis of host–pathogen interactions and our model provides a unique opportunity to gain basic insights into how animals avoid contaminated food. This article is part of the Theo Murphy meeting issue ‘Evolution of pathogen and parasite avoidance behaviours’.

G-proteins: Fighting infection on two fronts

Animals have evolved multiple strategies for coping with the presence of pathogenic microbes. The best characterized is the immune response where animals activate their physical and cellular defenses to respond to invading microorganisms. However, behavioral changes can also be triggered by exposure to microbes and play an important role in defending many species, including humans, from pathogen attack. In our recent study we demonstrate that, following infection, C. elegans uses the same G-protein signaling pathway in neurons and epithelial cells to coordinate avoidance behaviors and immune responses. Coordination of these responses allows animals to mount an immune response to the immediate threat while simultaneously taking action to remove the pathogen, however, the complicated nature of the mammalian brain and immune system has made it difficult to identify the molecular mechanisms mediating these interactions. With its simple, well described, nervous system and a rapidly growing understanding of its immune system, C. elegans has emerged as an excellent model to study the mechanisms by which animals recognize pathogens and coordinate behavioral and immune responses to infection.

Activation of RHO-1 in cholinergic motor neurons competes with dopamine signalling to control locomotion

The small GTPase RhoA plays a crucial role in the regulation of neuronal signalling to generate behaviour. In the developing nervous system RhoA is known to regulate the actin cytoskeleton, however the effectors of RhoA-signalling in adult neurons remain largely unidentified. We have previously shown that activation of the RhoA ortholog (RHO-1) in C. elegans cholinergic motor neurons triggers hyperactivity of these neurons and loopy locomotion with exaggerated body bends. This is achieved in part through increased diacylglycerol (DAG) levels and the recruitment of the synaptic vesicle protein UNC-13 to synaptic release sites, however other pathways remain to be identified. Dopamine, which is negatively regulated by the dopamine re-uptake transporter (DAT), has a central role in modulating locomotion in both humans and C. elegans. In this study we identify a new pathway in which RHO-1 regulates locomotory behaviour by repressing dopamine signalling, via DAT-1, linking these two pathways together. We observed an upregulation of dat-1 expression when RHO-1 is activated and show that loss of DAT-1 inhibits the loopy locomotion phenotype caused by RHO-1 activation. Reducing dopamine signalling in dat-1 mutants through mutations in genes involved in dopamine synthesis or in the dopamine receptor DOP-1 restores the ability of RHO-1 to trigger loopy locomotion in dat-1 mutants. Taken together, we show that negative regulation of dopamine signalling via DAT-1 is necessary for the neuronal RHO-1 pathway to regulate locomotion.

From head to tail it's a 2 way street for neuro-immune communication

Animals need to be able to rapidly and effectively respond to changes in their external and internal environment. To achieve this the nervous and immune systems need to coordinate their responses, integrating multiple cues including presence of potential pathogens, and availability of food. In our recent study 1 we demonstrate that signaling by sensory neurons in the head using the classical neurotransmitter serotonin can negatively regulate the rectal epithelial immune response upon infection of C. elegans with the naturally occurring bacterial pathogen Microbacterium nematophilum (M. nematophilum). The complicated nature of the mammalian brain and immune system has made it difficult to identify the molecular mechanisms mediating these interactions. With its simple, well described, nervous system and a rapidly growing understanding of its immune system, C. elegans has emerged as an excellent model to study the mechanisms by which animals recognize pathogens and coordinate behavioral and cellular immune responses to infection.