Multiphoton imaging of neural structure and activity in Drosophila through the intact cuticle

Aragon, Max Jameson; Mok, Aaron T.; Shea, Jamien; Wang, Mengran; Kim, Haein; Barkdull, Nathan; Xu, Chris; Yapici, Nilay

doi:10.7554/elife.69094

Cited by 19 publications

(17 citation statements)

References 53 publications

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“…A low-cost option is similarly available by substituting a DC stepper motor and a driver with microstepping capability to permit smooth acceleration. Both options would also permit compatibility with current platforms for in vivo imaging in Drosophila ( Aragon et al, 2022 ) and Caenorhabditis ( Smith et al, 2022 ). Naturally, our approach is compatible with head-mounted microscopes ( Aharoni and Hoogland, 2019 ; Zong et al, 2022 ) and stably mounted high-density probes of electrical activity ( Steinmetz et al, 2021 ) in rodents.…”

Section: Discussionmentioning

confidence: 99%

Tilt in Place Microscopy: a Simple, Low-Cost Solution to Image Neural Responses to Body Rotations

et al. 2022

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Animals use information about gravity and other destabilizing forces to balance and navigate through their environment. Measuring how brains respond to these forces requires considerable technical knowledge and/or financial resources. We present a simple alternative—Tilt In Place Microscopy (TIPM), a low-cost and noninvasive way to measure neural activity following rapid changes in body orientation. Here, we used TIPM to study vestibulospinal neurons in larval zebrafish during and immediately after roll tilts. Vestibulospinal neurons responded with reliable increases in activity that varied as a function of ipsilateral tilt amplitude. TIPM differentiated tonic (i.e., sustained tilt) from phasic responses, revealing coarse topography of stimulus sensitivity in the lateral vestibular nucleus. Neuronal variability across repeated sessions was minor relative to trial-to-trial variability, allowing us to use TIPM for longitudinal studies of the same neurons across two developmental time points. There, we observed global increases in response strength and systematic changes in the neural representation of stimulus direction. Our data extend classical characterization of the body tilt representation by vestibulospinal neurons and establish the utility of TIPM to study the neural basis of balance, especially in developing animals.SIGNIFICANCE STATEMENTVestibular sensation influences everything from navigation to interoception. Here, we detail a straightforward, validated, and nearly universal approach to image how the nervous system senses and responds to body tilts. We use our new method to replicate and expand on past findings of tilt sensing by a conserved population of spinal-projecting vestibular neurons. The simplicity and broad compatibility of our approach will democratize the study of the response of the brain to destabilization, particularly across development.

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Section: Discussionmentioning

confidence: 99%

Tilt in Place Microscopy: a Simple, Low-Cost Solution to Image Neural Responses to Body Rotations

et al. 2022

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“…A low-cost option is similarly available by substituting a DC stepper motor and a driver with micro-stepping capability to permit smooth acceleration. Both options would also permit compatibility with current platforms for in vivo imaging in Drosophila (Aragon et al, 2022) and Caenorhabditis (Smith et al, 2022). Naturally, our approach is compatible with head-mounted microscopes (Aharoni and Hoogland, 2019; Zong et al, 2022) and stably-mounted high-density probes of electrical activity (Steinmetz et al, 2021) in rodents.…”

Section: Discussionmentioning

confidence: 99%

Tilt In Place Microscopy (TIPM): a simple, low-cost solution to image neural responses to body rotations

Hamling

Zhu

Auer

et al. 2022

Preprint

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Animals use information about gravity and other destabilizing forces to balance and navigate through their environment. Measuring how brains respond to these forces requires considerable technical knowledge and/or financial resources. We present a simple alternative: Tilt In Place Microscopy (TIPM). TIPM is a low-cost and non-invasive way to measure neural activity following rapid changes in body orientation. Here we used TIPM to study vestibulospinal neurons in larval zebrafish during and immediately after roll tilts. Vestibulospinal neurons responded with reliable increases in activity that varied as a function of ipsilateral tilt amplitude. TIPM differentiated tonic (i.e. sustained tilt) from phasic responses, revealing coarse topography of stimulus sensitivity in the lateral vestibular nucleus. Neuronal variability across repeated sessions was minor relative to trial-to-trial variability, allowing us to use TIPM for longitudinal studies of the same neurons across two developmental timepoints. There, we observed global increases in response strength, and systematic changes in the neural representation of stimulus direction. Our data extend classical characterization of the body tilt representation by vestibulospinal neurons and establish TIPM's utility to study the neural basis of balance, especially in developing animals.

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“…The head cuticle was cut with a 16-gauge needle, and the air sacks, fat bodies, and trachea around the imaging window were removed using forceps. During all functional imaging experiments, flies stood or walked on an air-suspended, spherical treadmill (Aragon et al, 2022). Fly behavior was captured at 30 fps under IR illumination using two cameras (FLIR Blackfly-S, BFS-U3-13Y3C, and BFS-U3-16S2M) and the SpinView software (FLIR).…”

Section: Methods Detailsmentioning

confidence: 99%

“…During all functional imaging experiments, flies stood or walked on an air-suspended, spherical treadmill (Aragon et al, 2022). Fly behavior was captured at 30 fps under IR illumination using two cameras (FLIR Blackfly-S, BFS-U3-13Y3C, and BFS-U3-16S2M) and the SpinView software (FLIR).…”

Section: Author Contributionsmentioning

confidence: 99%

GABA-mediated inhibition in visual feedback neurons fine-tunesDrosophilamale courtship

Mabuchi

Cui

Xie

et al. 2023

Preprint

Self Cite

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Vision is critical for the regulation of mating behaviors in many species. Here, we discovered that the Drosophila ortholog of human GABAA-receptor-associated protein (GABARAP) is required to fine-tune male courtship by modulating the activity of visual feedback neurons, lamina tangential cells (Lat). GABARAP is a ubiquitin-like protein that regulates cell-surface levels of GABAA receptors. Knocking down GABARAP or GABAA receptors in Lat neurons or hyperactivating them induces male courtship toward other males. Inhibiting Lat neurons, on the other hand, delays copulation by impairing the ability of males to follow females. Remarkably, the human ortholog of Drosophila GABARAP restores function in Lat neurons. Using in vivo two-photon imaging and optogenetics, we show that Lat neurons are functionally connected to neural circuits that mediate visually-guided courtship pursuits in males. Our work reveals a novel physiological role for GABARAP in fine-tuning the activity of a visual circuit that tracks a mating partner during courtship.

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Multiphoton imaging of neural structure and activity in Drosophila through the intact cuticle

Cited by 19 publications

References 53 publications

Tilt in Place Microscopy: a Simple, Low-Cost Solution to Image Neural Responses to Body Rotations

Tilt in Place Microscopy: a Simple, Low-Cost Solution to Image Neural Responses to Body Rotations

Tilt In Place Microscopy (TIPM): a simple, low-cost solution to image neural responses to body rotations

GABA-mediated inhibition in visual feedback neurons fine-tunesDrosophilamale courtship

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