A Semi-supervised Pipeline for Accurate Neuron Segmentation with Fewer Ground Truth Labels.

Recent advancements in two-photon calcium imaging have enabled scientists to record the activity of thousands of neurons with cellular resolution. This scope of data collection is crucial to understanding the next generation of neuroscience questions, but analyzing these large recordings requires automated methods for neuron segmentation. Supervised methods for neuron segmentation achieve state of-the-art accuracy and speed but currently require large amounts of manually generated ground truth training labels. We reduced the required number of training labels by designing a semi-supervised pipeline. Our pipeline used neural network ensembling to generate pseudolabels to train a single shallow U-Net. We tested our method on three publicly available datasets and compared our performance to three widely used segmentation methods. Our method outperformed other methods when trained on a small number of ground truth labels and could achieve state-of-the-art accuracy after training on approximately a quarter of the number of ground truth labels as supervised methods. When trained on many ground truth labels, our pipeline attained higher accuracy than that of state-of-the-art methods. Overall, our work will help researchers accurately process large neural recordings while minimizing the time and effort needed to generate manual labels.

A single neuron in C. elegans orchestrates multiple motor outputs through parallel modes of transmission.

Animals generate a wide range of highly coordinated motor outputs, which allows them to execute purposeful behaviors. Individual neurons in the circuits that generate behaviors have a remarkable capacity for flexibility as they exhibit multiple axonal projections, transmitter systems, and modes of neural activity. How these multi-functional properties of neurons enable the generation of adaptive behaviors remains unknown. Here, we show that the HSN neuron in C. elegans evokes multiple motor programs over different timescales to enable a suite of behavioral changes during egg laying. Using HSN activity perturbations and in vivo calcium imaging, we show that HSN acutely increases egg laying and locomotion while also biasing the animals toward low-speed dwelling behavior over minutes. The acute effects of HSN on egg laying and high-speed locomotion are mediated by separate sets of HSN transmitters and different HSN axonal compartments. The long-lasting effects on dwelling are mediated in part by HSN release of serotonin, which is taken up and re-released by NSM, another serotonergic neuron class that directly evokes dwelling. Our results show how the multi-functional properties of a single neuron allow it to induce a coordinated suite of behaviors and also reveal that neurons can borrow serotonin from one another to control behavior.

Identifying properties of pattern completion neurons in a computational model of the visual cortex.

Neural ensembles are found throughout the brain and are believed to underlie diverse cognitive functions including memory and perception. Methods to activate ensembles precisely, reliably, and quickly are needed to further study the ensembles' role in cognitive processes. Previous work has found that ensembles in layer 2/3 of the visual cortex (V1) exhibited pattern completion properties: ensembles containing tens of neurons were activated by stimulation of just two neurons. However, methods that identify pattern completion neurons are underdeveloped. In this study, we optimized the selection of pattern completion neurons in simulated ensembles. We developed a computational model that replicated the connectivity patterns and electrophysiological properties of layer 2/3 of mouse V1. We identified ensembles of excitatory model neurons using K-means clustering. We then stimulated pairs of neurons in identified ensembles while tracking the activity of the entire ensemble. Our analysis of ensemble activity quantified a neuron pair's power to activate an ensemble using a novel metric called pattern completion capability (PCC) based on the mean pre-stimulation voltage across the ensemble. We found that PCC was directly correlated with multiple graph theory parameters, such as degree and closeness centrality. To improve selection of pattern completion neurons in vivo, we computed a novel latency metric that was correlated with PCC and could potentially be estimated from modern physiological recordings. Lastly, we found that stimulation of five neurons could reliably activate ensembles. These findings can help researchers identify pattern completion neurons to stimulate in vivo during behavioral studies to control ensemble activation.

A single neuron in C. elegans orchestrates multiple motor outputs through parallel modes of transmission.

Animals generate a wide range of highly coordinated motor outputs, which allows them to execute purposeful behaviors. Individual neuron classes in the circuits that generate behavior have a remarkable capacity for flexibility, as they exhibit multiple axonal projections, transmitter systems, and modes of neural activity. How these multi-functional properties of neurons enable the generation of highly coordinated behaviors remains unknown. Here we show that the HSN neuron in C. elegans evokes multiple motor programs over different timescales to enable a suite of behavioral changes during egg-laying. Using HSN activity perturbations and in vivo calcium imaging, we show that HSN acutely increases egg-laying and locomotion while also biasing the animals towards low-speed dwelling behavior over longer timescales. The acute effects of HSN on egg-laying and high-speed locomotion are mediated by separate sets of HSN transmitters and different HSN axonal projections. The long-lasting effects on dwelling are mediated by HSN release of serotonin that is taken up and re-released by NSM, another serotonergic neuron class that directly evokes dwelling. Our results show how the multi-functional properties of a single neuron allow it to induce a coordinated suite of behaviors and also reveal for the first time that neurons can borrow serotonin from one another to control behavior.

A neural circuit for flexible control of persistent behavioral states.

To adapt to their environments, animals must generate behaviors that are closely aligned to a rapidly changing sensory world. However, behavioral states such as foraging or courtship typically persist over long time scales to ensure proper execution. It remains unclear how neural circuits generate persistent behavioral states while maintaining the flexibility to select among alternative states when the sensory context changes. Here, we elucidate the functional architecture of a neural circuit controlling the choice between roaming and dwelling states, which underlie exploration and exploitation during foraging in C. elegans. By imaging ensemble-level neural activity in freely moving animals, we identify stereotyped changes in circuit activity corresponding to each behavioral state. Combining circuit-wide imaging with genetic analysis, we find that mutual inhibition between two antagonistic neuromodulatory systems underlies the persistence and mutual exclusivity of the neural activity patterns observed in each state. Through machine learning analysis and circuit perturbations, we identify a sensory processing neuron that can transmit information about food odors to both the roaming and dwelling circuits and bias the animal towards different states in different sensory contexts, giving rise to context-appropriate state transitions. Our findings reveal a potentially general circuit architecture that enables flexible, sensory-driven control of persistent behavioral states.