Transsynaptic labeling and transcriptional control of zebrafish neural circuits.

Deciphering the connectome, the ensemble of synaptic connections that underlie brain function, is a central goal of neuroscience research. Here we report the in vivo mapping of connections between presynaptic and postsynaptic partners in zebrafish, by adapting the trans-Tango genetic approach that was first developed for anterograde transsynaptic tracing in Drosophila. Neural connections were visualized between synaptic partners in larval retina, brain and spinal cord and followed over development.

Cell-type-specific origins of locomotor rhythmicity at different speeds in larval zebrafish.

Different speeds of locomotion require heterogeneous spinal populations, but a common mode of rhythm generation is presumed to exist. Here, we explore the cellular versus synaptic origins of spinal rhythmicity at different speeds by performing electrophysiological recordings from premotor excitatory interneurons in larval zebrafish. Chx10-labeled V2a neurons are divided into at least two morphological subtypes proposed to play distinct roles in timing and intensity control.

Cell-type-specific origins of locomotor rhythmicity at different speeds in larval zebrafish.

Different speeds of locomotion require heterogeneous spinal populations, but a common mode of rhythm generation is presumed to exist. Here, we explore the cellular versus synaptic origins of spinal rhythmicity at different speeds by performing electrophysiological recordings from premotor excitatory interneurons in larval zebrafish. Chx10-labeled V2a neurons are divided into at least two morphological subtypes proposed to play distinct roles in timing and intensity control.

Spinal Basis of Direction Control during Locomotion in Larval Zebrafish.

Navigation requires steering and propulsion, but how spinal circuits contribute to direction control during ongoing locomotion is not well understood. Here, we use drifting vertical gratings to evoke directed "fictive" swimming in intact but immobilized larval zebrafish while performing electrophysiological recordings from spinal neurons. We find that directed swimming involves unilateral changes in the duration of motor output and increased recruitment of motor neurons, without impacting the timing of spiking across or along the body.

Mixed synapses reconcile violations of the size principle in zebrafish spinal cord.

Mixed electrical-chemical synapses potentially complicate electrophysiological interpretations of neuronal excitability and connectivity. Here, we disentangle the impact of mixed synapses within the spinal locomotor circuitry of larval zebrafish. We demonstrate that soma size is not linked to input resistance for interneurons, contrary to the biophysical predictions of the 'size principle' for motor neurons.

Voltage imaging and spinal circuits get along swimmingly.

Voltage imaging promises to unite optical and electrical approaches to accelerate circuit discovery. In this issue of Neuron, Böhm et al. (2022) use voltage imaging to explore the structure and functional dynamics of spinal excitatory interneurons in larval zebrafish and reveal the role of V3 neurons in adaptive locomotor control.

Locomotor control: Inhibiting actions locally and senses globally.

Locomotion requires the segmental coordination of activity along the body. A new study in zebrafish reveals that spinal inhibitory interneurons are wired to execute different functions depending on whether their targets are nearby or further away.

Intersection of motor volumes predicts the outcome of ambush predation of larval zebrafish.

Escape maneuvers are key determinants of animal survival and are under intense selection pressure. A number of escape maneuver parameters contribute to survival, including response latency, escape speed and direction. However, the relative importance of these parameters is context dependent, suggesting that interactions between parameters and predatory context determine the likelihood of escape success.

Orderly compartmental mapping of premotor inhibition in the developing zebrafish spinal cord.

In vertebrates, faster movements involve the orderly recruitment of different types of spinal motor neurons. However, it is not known how premotor inhibitory circuits are organized to ensure alternating motor output at different movement speeds. We found that different types of commissural inhibitory interneurons in zebrafish form compartmental microcircuits during development that align inhibitory strength and recruitment order.

Integration of Swimming-Related Synaptic Excitation and Inhibition by olig2 Eurydendroid Neurons in Larval Zebrafish Cerebellum.

The cerebellum influences motor control through Purkinje target neurons, which transmit cerebellar output. Such output is required, for instance, for larval zebrafish to learn conditioned fictive swimming. The output cells, called eurydendroid neurons (ENs) in teleost fish, are inhibited by Purkinje cells and excited by parallel fibers.

Reconciling the functions of even-skipped interneurons during crawling, swimming, and walking.

In all bilaterally symmetric animals, movements across the body are coordinated by interneurons that traverse the midline. Recent work is beginning to tease apart the functional complexity of interneurons labeled by the homeodomain transcription factor even-skipped, which provide a phylogenetically-conserved source of commissural excitation during locomotion in both vertebrates and invertebrates. Here we review recent studies of the roles of even-skipped neurons during locomotion in flies (EL neurons), fishes, frogs, and mice (V0v neurons).

Hierarchical control of locomotion by distinct types of spinal V2a interneurons in zebrafish.

In all vertebrates, excitatory spinal interneurons execute dynamic adjustments in the timing and amplitude of locomotor movements. Currently, it is unclear whether interneurons responsible for timing control are distinct from those involved in amplitude control. Here, we show that in larval zebrafish, molecularly, morphologically and electrophysiologically distinct types of V2a neurons exhibit complementary patterns of connectivity.

Central and peripheral innervation patterns of defined axial motor units in larval zebrafish.

Spinal motor neurons and the peripheral muscle fibers they innervate form discrete motor units that execute movements of varying force and speed. Subsets of spinal motor neurons also exhibit axon collaterals that influence motor output centrally. Here, we have used in vivo imaging to anatomically characterize the central and peripheral innervation patterns of axial motor units in larval zebrafish.

Integrated one- and two-photon scanned oblique plane illumination (SOPi) microscopy for rapid volumetric imaging.

Versatile, sterically accessible imaging systems capable of in vivo rapid volumetric functional and structural imaging deep in the brain continue to be a limiting factor in neuroscience research. Towards overcoming this obstacle, we present integrated one- and two-photon scanned oblique plane illumination (SOPi, /sōpī/) microscopy which uses a single front-facing microscope objective to provide light-sheet scanning based rapid volumetric imaging capability at subcellular resolution. Our planar scan-mirror based optimized light-sheet architecture allows for non-distorted scanning of volume samples, simplifying accurate reconstruction of the imaged volume.

Visual Threat Assessment and Reticulospinal Encoding of Calibrated Responses in Larval Zebrafish.

All visual animals must decide whether approaching objects are a threat. Our current understanding of this process has identified a proximity-based mechanism where an evasive maneuver is triggered when a looming stimulus passes a subtended visual angle threshold. However, some escape strategies are more costly than others, and so it would be beneficial to additionally encode the level of threat conveyed by the predator's approach rate to select the most appropriate response.

Distinct responses of Purkinje neurons and roles of simple spikes during associative motor learning in larval zebrafish.

To study cerebellar activity during learning, we made whole-cell recordings from larval zebrafish Purkinje cells while monitoring fictive swimming during associative conditioning. Fish learned to swim in response to visual stimulation preceding tactile stimulation of the tail. Learning was abolished by cerebellar ablation.

Peeling back the layers of locomotor control in the spinal cord.

Vertebrate locomotion is executed by networks of neurons within the spinal cord. Here, we describe recent advances in our understanding of spinal locomotor control provided by work using optical and genetic approaches in mice and zebrafish. In particular, we highlight common observations that demonstrate simplification of limb and axial motor pool coordination by spinal network modularity, differences in the deployment of spinal modules at increasing speeds of locomotion, and functional hierarchies in the regulation of locomotor rhythm and pattern.

Neuromodulation: letting sources of spinal dopamine speak for themselves.

A recent study of dopaminergic neurons in the brain of larval zebrafish has important implications for interpreting the natural actions of neuromodulators in the spinal cord.

Systematic shifts in the balance of excitation and inhibition coordinate the activity of axial motor pools at different speeds of locomotion.

An emerging consensus from studies of axial and limb networks is that different premotor populations are required for different speeds of locomotion. An important but unresolved issue is why this occurs. Here, we perform voltage-clamp recordings from axial motoneurons in larval zebrafish during "fictive" swimming to test the idea that systematic differences in the biophysical properties of axial motoneurons are associated with differential tuning in the weight and timing of synaptic drive, which would help explain premotor population shifts.

Selective responses to tonic descending commands by temporal summation in a spinal motor pool.

Motor responses of varying intensities rely on descending commands to heterogeneous pools of motoneurons. In vertebrates, numerous sources of descending excitatory input provide systematically more drive to progressively less excitable spinal motoneurons. While this presumably facilitates simultaneous activation of motor pools, it is unclear how selective patterns of recruitment could emerge from inputs weighted this way.

Modular organization of axial microcircuits in zebrafish.

Locomotion requires precise control of spinal networks. In tetrapods and bipeds, dynamic regulation of locomotion is simplified by the modular organization of spinal limb circuits, but it is not known whether their predecessors, fish axial circuits, are similarly organized. Here, we demonstrate that the larval zebrafish spinal cord contains distinct, parallel microcircuits for independent control of dorsal and ventral musculature on each side of the body.

Differences in the morphology of spinal V2a neurons reflect their recruitment order during swimming in larval zebrafish.

Networks of neurons in spinal cord generate locomotion. However, little is known about potential differences in network architecture that underlie the production of varying speeds of movement. In larval zebrafish, as swimming speed increases, Chx10-positive V2a excitatory premotor interneurons are activated from ventral to dorsal in a topographic pattern that parallels axial motoneuron recruitment.

Speed control: spinal interneurons with crossed purposes.

A recent study has revealed that different populations of commissural spinal interneurons ensure limb alternation at different speeds of locomotion.

Optogenetics: illuminating sources of locomotor drive.

A recent study has used optogenetics to identify the source of excitatory drive for locomotion in zebrafish, revealing unexpected differences in the command signals from hindbrain to spinal cord.