SAMPL is a high-throughput solution to study unconstrained vertical behavior in small animals

Zhu, Yunlu; Auer, Franziska; Gelnaw, Hannah; Davis, Samantha N.; Hamling, Kyla R.; May, Christina E.; Ahamed, Hassan; Ringstad, Niels; Nagel, Katherine I.; Schoppik, David

doi:10.1016/j.celrep.2023.112573

Cited by 15 publications

(14 citation statements)

References 130 publications

(124 reference statements)

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“…We used our Scalable Apparatus to Measure Posture and Locomotion (SAMPL) to measure posture and locomotion in freely swimming zebrafish 20 . SAMPL is a high-throughput videographic approach that measures kinematic parameters of posture and locomotion from fish swimming in a predominantly vertical arena that encourages navigation in depth (Figures 2A and 2B).…”

Section: Resultsmentioning

confidence: 99%

“…the presence/absence of the elavl3:h2B-GCaMP6f) allele). Because of background variation 20 , all comparisons were restricted to control vs. experimental groups within an experimental paradigm over the same time period. Across our datasets (Tables 1 to 4) we did not observe meaningful differences between the control and experimental groups in the pre-manipulation period.…”

Section: Resultsmentioning

confidence: 99%

“…We hypothesized that their activity would be modulated by body tilts in the pitch axis. We used Tilt In Place Microscopy (TIPM) 20 to measure the response of individual Purkinje cells (Figure 5A) to rapid pitch tilts. Briefly, fish are mounted on a mirror galvanometer and rapidly rotated to eccentric angles (Figure 5B, ± 30°nose-up/nose-down).…”

Section: Resultsmentioning

confidence: 99%

“…All behavior was measured using the Scalable Apparatus for Measuring Posture and Locomotion (SAMPL) apparatus, consisting of a chamber where larvae could swim freely, an infrared illuminator, a camera, and software to process video in real time. A comprehensive description of the apparatus is contained in 20 . Here we briefly describe the specific details of our experiments.…”

Section: Methodsmentioning

confidence: 99%

“…For example, larval zebrafish balance in the pitch axis (nose-up/nose-down) by timing locomotion to countermand gravity-induced destabilization 17,18 and by coordinated use of paired appendages (fins) and axial musculature (trunk) 19 . The small size and rapid development of the larval zebrafish allow high-throughput measurements of these movements from freely swimming subjects 20 .…”

Section: Introductionmentioning

confidence: 99%

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Cerebellar Purkinje Cells Control Posture in Larval Zebrafish (Danio rerio)

Auer,

Nardone,

Matsuda

et al. 2023

Preprint

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Cerebellar dysfunction leads to postural instability. Recent work in freely moving rodents has transformed investigations of cerebellar contributions to posture. However, the combined complexity of terrestrial locomotion and the rodent cerebellum motivate development of new approaches to perturb cerebellar function in simpler vertebrates. Here, we used a powerful chemogenetic tool (TRPV1/capsaicin) to define the role of Purkinje cells — the output neurons of the cerebellar cortex — as larval zebrafish swam freely in depth. We achieved both bidirectional control (activation and ablation) of Purkinje cells while performing quantitative high-throughput assessment of posture and locomotion. Activation disrupted postural control in the pitch (nose-up/nose-down) axis. Similarly, ablations disrupted pitch-axis posture and fin-body coordination responsible for climbs. Postural disruption was more pronounced in older larvae, offering a window into emergent roles for the developing cerebellum in the control of posture. Finally, we found that activity in Purkinje cells could individually and collectively encode tilt direction, a key feature of postural control neurons. Our findings delineate an expected role for the cerebellum in postural control and vestibular sensation in larval zebrafish, establishing the validity of TRPV1/capsaicin-mediated perturbations in a simple, genetically-tractable vertebrate. Moreover, by comparing the contributions of Purkinje cell ablations to posture in time, we uncover signatures of emerging cerebellar control of posture across early development. This work takes a major step towards understanding an ancestral role of the cerebellum in regulating postural maturation.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Cerebellar Purkinje Cells Control Posture in Larval Zebrafish (Danio rerio)

Auer,

Nardone,

Matsuda

et al. 2023

Preprint

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Organization of vestibular circuits for postural control in zebrafish

Liu,

Bagnall

2023

Current Opinion in Neurobiology

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pi_tailtrack: A compact, inexpensive, and open-source behaviour-tracking system for head-restrained zebrafish

Randlett

2023

Preprint

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Quantifying animal behavior during microscopy is crucial to associate optically recorded neural activity with behavioural outputs and states. Here I describe an imaging and tracking system for head-restrained larval zebrafish compatible with functional microscopy. This system is based on the Raspberry Pi computer, Pi NoIR camera, and open-source software for the real-time tail segmentation and skeletonization of the zebrafish tail at over 100hz. This allows for precise and long-term analyses of swimming behaviour, that can be related to functional signals recorded in individual neurons. This system offers a simple but performant solution for quantifying the behavior of head-restrained larval zebrafish, which can be built for 340 euro.

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SAMPL is a high-throughput solution to study unconstrained vertical behavior in small animals

Cited by 15 publications

References 130 publications

Cerebellar Purkinje Cells Control Posture in Larval Zebrafish (Danio rerio)

Cerebellar Purkinje Cells Control Posture in Larval Zebrafish (Danio rerio)

Organization of vestibular circuits for postural control in zebrafish

pi_tailtrack: A compact, inexpensive, and open-source behaviour-tracking system for head-restrained zebrafish

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