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bioRxiv · 10.1101/2024.07.28.605489

Using DeepLabCut-Live to probe state dependent neural circuits of behavior with closed-loop optogenetic stimulation

Gonzalez, M.·Gradwell, M. A.·Thackray, J. K.·Patel, K. R.·Temkar, K. K.·Abraira, V. E. G.
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Abstract

BackgroundClosed-loop behavior paradigms enable us to dissect the state-dependent neural circuits underlying behavior in real-time. However, studying context-dependent locomotor perturbations has been challenging due to limitations in molecular tools and techniques for real-time manipulation of spinal cord circuits. New MethodWe developed a novel closed-loop optogenetic stimulation paradigm that utilizes DeepLabCut-Live pose estimation to manipulate primary sensory afferent activity at specific phases of the locomotor cycle in mice. A compact DeepLabCut model was trained to track hindlimb kinematics in real-time and integrated into the Bonsai visual programming framework. This allowed an LED to be triggered to photo-stimulate sensory neurons expressing channelrhodopsin at user-defined pose-based criteria, such as during the stance or swing phase. ResultsOptogenetic activation of nociceptive TRPV1+ sensory neurons during treadmill locomotion reliably evoked paw withdrawal responses. Photoactivation during stance generated a brief withdrawal, while stimulation during swing elicited a prolonged response likely engaging stumbling corrective reflexes. Comparison with Existing Methods: This new method allows for high spatiotemporal precision in manipulating spinal circuits based on the phase of the locomotor cycle. Unlike previous approaches, this closed-loop system can control for the state-dependent nature of sensorimotor responses during locomotion. ConclusionsIntegrating DeepLabCut-Live with optogenetics provides a powerful new approach to dissect the context-dependent role of sensory feedback and spinal interneurons in modulating locomotion. This technique opens new avenues for uncovering the neural substrates of state-dependent behaviors and has broad applicability for studies of real-time closed-loop manipulation based on pose estimation. ManuscriptO_ST_ABSHighlightsC_ST_ABSO_LIClosed-loop system probes state-dependent behaviors at pose-modulated instances C_LIO_LIBonsai integrates DeepLabCut models for real-time pose estimation during locomotion C_LIO_LIPhase-dependent TRPV1+ sensory afferent photostimulation elicits context-specific withdrawal responses C_LI

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Gonzalez, M., Gradwell, M. A., Thackray, J. K., Patel, K. R., Temkar, K. K., Abraira, V. E. G.. 2024-07-29. Using DeepLabCut-Live to probe state dependent neural circuits of behavior with closed-loop optogenetic stimulation. https://doi.org/10.1101/2024.07.28.605489

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