65 research outputs found

    The Neural Circuitry Of Social Behavior In C. elegans

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    Most animal species, from simple invertebrates to complex mammals, require behavioral mechanisms to communicate with and respond to conspecifics, whether to mate, to assess predatory danger, or evaluate the nutritional quality of the surrounding environment. Understanding the molecular and cellular underpinnings of these social behaviors remains a central challenge in neurobiology. I used the nematode C. elegans as a model system to study the genetics and neural circuitry that underlie social behavior. First, I evaluated the behavioral responses of C. elegans to a nematode extract (deathmone), which served as a model for alarm pheromones in other animal species (chapter 2). Worms showed acute avoidance of deathmone, and reduced their exploration when cultivated on it, a behavior termed “dwelling.†I combined chemical analysis, laser ablation studies, and genetic studies to identify the sensory neurons and molecular signaling pathways that promote dwelling in response to deathmone. Second, I investigated the neuronal substrates responsible for social feeding, a behavior in which certain strains of C. elegans display high lomocotory speeds, accumulate on the border of bacterial food lawns, and aggregate into groups. A low activity or null allele of the neuropeptide y receptor homologue npr-1 promotes social feeding, while a high activity form—which is found in the wild-type N2 strain—promotes solitary behavior1. Expression of a high-activity npr-1 cDNA specifically in the interneuron RMG converted npr-1 loss-of-function mutants from social feeders into solitary ones. The RMG neurons are gap junctional hubs that electrically couple the sensory neurons URX, ASH, and ADL—all previously implicated in social feeding—and the pheromone-sensing neuron ASK, suggesting that social feeding and pheromone responses may be related. Indeed, npr-1 social feeders are attracted to ascarosides, while N2 solitary feeders are repelled, a behavioral difference that is dependent on RMG function. Calcium imaging of ASK and its postsynaptic partner AIA demonstrated that RMG promotes signaling from ASK to AIA. Taken together, these data provide a common neural circuitry for social behaviors in C. elegans, and offer some insights into the molecular mechanisms of their regulation

    Comprehensive Classification of Retinal Bipolar Neurons by Single-Cell Transcriptomics

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    Patterns of gene expression can be used to characterize and classify neuronal types. It is challenging, however, to generate taxonomies that fulfill the essential criteria of being comprehensive, harmonizing with conventional classification schemes, and lacking superfluous subdivisions of genuine types. To address these challenges, we used massively parallel single-cell RNA profiling and optimized computational methods on a heterogeneous class of neurons, mouse retinal bipolar cells (BCs). From a population of ∼25,000 BCs, we derived a molecular classification that identified 15 types, including all types observed previously and two novel types, one of which has a non-canonical morphology and position. We validated the classification scheme and identified dozens of novel markers using methods that match molecular expression to cell morphology. This work provides a systematic methodology for achieving comprehensive molecular classification of neurons, identifies novel neuronal types, and uncovers transcriptional differences that distinguish types within a class

    Neuromodulatory state and sex specify alternative behaviors through antagonistic synaptic pathways in C. elegans

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    SUMMARY Pheromone responses are highly context dependent. For example, the C. elegans pheromone ascaroside C9 (ascr#3) is repulsive to wild-type hermaphrodites, attractive to wild-type males, and usually neutral to ''social'' hermaphrodites with reduced activity of the npr-1 neuropeptide receptor gene. We show here that these distinct behavioral responses arise from overlapping push-pull circuits driven by two classes of pheromone-sensing neurons. The ADL sensory neurons detect C9 and, in wild-type hermaphrodites, drive C9 repulsion through their chemical synapses. In npr-1 mutant hermaphrodites, C9 repulsion is reduced by the recruitment of a gap junction circuit that antagonizes ADL chemical synapses. In males, ADL sensory responses are diminished; in addition, a second pheromone-sensing neuron, ASK, antagonizes C9 repulsion. The additive effects of these antagonistic circuit elements generate attractive, repulsive, or neutral pheromone responses. Neuronal modulation by circuit state and sex, and flexibility in synaptic output pathways, may permit small circuits to maximize their adaptive behavioral outputs

    Dissection of artifactual and confounding glial signatures by single-cell sequencing of mouse and human brain

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    A key aspect of nearly all single-cell sequencing experiments is dissociation of intact tissues into single-cell suspensions. While many protocols have been optimized for optimal cell yield, they have often overlooked the effects that dissociation can have on ex vivo gene expression. Here, we demonstrate that use of enzymatic dissociation on brain tissue induces an aberrant ex vivo gene expression signature, most prominently in microglia, which is prevalent in published literature and can substantially confound downstream analyses. To address this issue, we present a rigorously validated protocol that preserves both in vivo transcriptional profiles and cell-type diversity and yield across tissue types and species. We also identify a similar signature in postmortem human brain single-nucleus RNA-sequencing datasets, and show that this signature is induced in freshly isolated human tissue by exposure to elevated temperatures ex vivo. Together, our results provide a methodological solution for preventing artifactual gene expression changes during fresh tissue digestion and a reference for future deeper analysis of the potential confounding states present in postmortem human samples

    The Human Cell Atlas White Paper

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    The Human Cell Atlas (HCA) will be made up of comprehensive reference maps of all human cells - the fundamental units of life - as a basis for understanding fundamental human biological processes and diagnosing, monitoring, and treating disease. It will help scientists understand how genetic variants impact disease risk, define drug toxicities, discover better therapies, and advance regenerative medicine. A resource of such ambition and scale should be built in stages, increasing in size, breadth, and resolution as technologies develop and understanding deepens. We will therefore pursue Phase 1 as a suite of flagship projects in key tissues, systems, and organs. We will bring together experts in biology, medicine, genomics, technology development and computation (including data analysis, software engineering, and visualization). We will also need standardized experimental and computational methods that will allow us to compare diverse cell and tissue types - and samples across human communities - in consistent ways, ensuring that the resulting resource is truly global. This document, the first version of the HCA White Paper, was written by experts in the field with feedback and suggestions from the HCA community, gathered during recent international meetings. The White Paper, released at the close of this yearlong planning process, will be a living document that evolves as the HCA community provides additional feedback, as technological and computational advances are made, and as lessons are learned during the construction of the atlas
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