A new platform for long-term tracking and recording of neural activity and simultaneous optogenetic control in freely behaving Caenorhabditis elegans

Gengyo-Ando, Keiko; Kagawa-Nagamura, Yuko; Ohkura, Masamichi; Fei, Xianfeng; Chen, Min; Hashimoto, Koichi; Nakai, Junichi

doi:10.1016/j.jneumeth.2017.05.017

Cited by 12 publications

(19 citation statements)

References 66 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Calcium imaging of RME and RIM in freely behaving animals was performed using the autotracking imaging system ICaST as described previously (Additional file 1 : Figure S1) [ 27 ]. Briefly, to automatically track a moving animal in the field of view of the laser-scanning microscope, infrared worm images were taken at 200 frames/s and image processing was conducted in real-time.…”

Section: Methodsmentioning

confidence: 99%

“…For optogenetic analysis, transgenic animals expressing channelrhodopsin-2 (ChR2) in ASH neurons were cultivated on NGM with all- trans -retinal (ATR) (Sigma-Aldrich Japan, KK, Tokyo) in the dark for 1–3 days before experiments. Simultaneous imaging and optogenetic control was performed as described previously [ 27 ]. To avoid inappropriate excitation of G-CaMP, blue light at 440 ± 10.5 nm, which is far from the excitation peak of G-CaMP7, was used for the photoactivation of ChR2.…”

Section: Methodsmentioning

confidence: 99%

“…To minimize optical crosstalk, the intensities of the 488 nm laser and 440 nm blue light were optimized. Detailed methods for controlling the light intensity are described in a previous report [ 27 ]. For calcium imaging of optogenetically evoked reversals and omega turns, photostimulation for 5–20 s was delivered after animals moved forward for at least 5 s.…”

Section: Methodsmentioning

confidence: 99%

“…Our preliminary calcium imaging data have suggested that RME are affected by tyramine (Gengyo-Ando et al, 2013; the 19th international worm meeting, University of California, Los Angeles). In the present study, to further understand the neuromodulation of RME, we examined the correlation between the calcium dynamics of RME and behavior using an autotracking imaging system, ICaST [ 27 ]. Our data demonstrate that tyramine alters calcium levels in RME neurons in a specific behavioral state.…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Role of tyramine in calcium dynamics of GABAergic neurons and escape behavior in Caenorhabditis elegans

et al. 2018

Self Cite

View full text Add to dashboard Cite

BackgroundTyramine, known as a “trace amine” in mammals, modulates a wide range of behavior in invertebrates; however, the underlying cellular and circuit mechanisms are not well understood. In the nematode Caenorhabditis elegans (C. elegans), tyramine affects key behaviors, including foraging, feeding, and escape responses. The touch-evoked backward escape response is often coupled with a sharp omega turn that allows the animal to navigate away in the opposite direction. Previous studies have showed that a metabotropic tyramine receptor, SER-2, in GABAergic body motor neurons controls deep body bending in omega turns. In this study, we focused on the role of tyramine in GABAergic head motor neurons. Our goal is to understand the mechanism by which tyraminergic signaling alters neural circuit activity to control escape behavior.ResultsUsing calcium imaging in freely moving C. elegans, we found that GABAergic RME motor neurons in the head had high calcium levels during forward locomotion but low calcium levels during spontaneous and evoked backward locomotion. This calcium decrease was also observed during the omega turn. Mutant analyses showed that tbh-1 mutants lacking only octopamine had normal calcium responses, whereas tdc-1 mutants lacking both tyramine and octopamine did not exhibit the calcium decrease in RME. This neuromodulation was mediated by SER-2. Moreover, tyraminergic RIM neuron activity was negatively correlated with RME activity in the directional switch from forward to backward locomotion. These results indicate that tyramine released from RIM inhibits RME via SER-2 signaling. The omega turn is initiated by a sharp head bend when the animal reinitiates forward movement. Interestingly, ser-2 mutants exhibited shallow head bends and often failed to execute deep-angle omega turns. The behavioral defect and the abnormal calcium response in ser-2 mutants could be rescued by SER-2 expression in RME. These results suggest that tyraminergic inhibition of RME is involved in the control of omega turns.ConclusionWe demonstrate that endogenous tyramine downregulates calcium levels in GABAergic RME motor neurons in the head via the tyramine receptor SER-2 during backward locomotion and omega turns. Our data suggest that this neuromodulation allows deep head bending during omega turns and plays a role in the escape behavior in C. elegans.Electronic supplementary materialThe online version of this article (10.1186/s40851-018-0103-1) contains supplementary material, which is available to authorized users.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Role of tyramine in calcium dynamics of GABAergic neurons and escape behavior in Caenorhabditis elegans

et al. 2018

Self Cite

View full text Add to dashboard Cite

show abstract

“…Through the development of new rhodopsins, first steps towards long-term optogenetics have been made. These newer genetic tools can be activated for minutes (Gengyo-Ando et al, 2017) or even up to 2 days (Schultheis et al, 2011). The longest optogenetic lifespan experiment to date lasted 2.5 hours (De Rosa et al, 2019).…”

Section: Introductionmentioning

confidence: 99%

The OptoGenBox - a device for long-term optogenetics inC. elegans

Busack

Jordan

Sapir

et al. 2020

Preprint

View full text Add to dashboard Cite

show abstract

Fast whole‐body motor neuron calcium imaging of freely moving Caenorhabditis elegans without coverslip pressed

Fang

Zhai

et al. 2021

Cytometry Pt A

View full text Add to dashboard Cite

Caenorhabditis elegans (C. elegans) is an ideal model organism for studying neuronal functions at the system level. This article develops a customized system for wholebody motor neuron calcium imaging of freely moving C. elegans without the coverslip pressed. Firstly, we proposed a fast centerline localization algorithm that could deal with most topology-variant cases costing only 6 ms for one frame, not only benefits for real-time localization but also for post-analysis. Secondly, we implemented a fulltime two-axis synchronized motion strategy by adaptively adjusting the motion parameters of two motors in every short-term motion step (~50 ms). Following the above motion tracking configuration, the tracking performance of our system has been demonstrated to completely support the high spatiotemporal resolution calcium imaging on whole-body motor neurons of wild-type (N2) worms as well as two mutants (unc-2, unc-9), even the instantaneous speed of worm moving without coverslip pressed was extremely up to 400 μm/s.

show abstract

A new platform for long-term tracking and recording of neural activity and simultaneous optogenetic control in freely behaving Caenorhabditis elegans

Cited by 12 publications

References 66 publications

Role of tyramine in calcium dynamics of GABAergic neurons and escape behavior in Caenorhabditis elegans

Role of tyramine in calcium dynamics of GABAergic neurons and escape behavior in Caenorhabditis elegans

The OptoGenBox - a device for long-term optogenetics inC. elegans

Fast whole‐body motor neuron calcium imaging of freely moving Caenorhabditis elegans without coverslip pressed

Contact Info

Product

Resources

About