Optogenetic manipulation of neuronal and cardiomyocyte functions in zebrafish using microbial rhodopsins and adenylyl cyclases
- Graduate School of Science, Nagoya University, Japan, Japan
- Graduate School of Bioagricultural Sciences, Nagoya University, Japan
- Institute for Advanced Research, Nagoya University, Japan
- Department of Life Science and Applied Chemistry, Nagoya Institute of Technology, Japan
Figures
Figure 1 with 1 supplement
Photocurrent properties of channelrhodopsins.
(A) Representative photocurrent traces of GtCCR4-3.0-EYFP (GtCCR4-3.0), GtCCR4-MT-P2A-TagCFP (GtCCR4-MT), KnChR, CrChR2[T159C], CoChR, and ChrimsonR. Electrophysiological recordings were performed. …
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Figure 1—source data 1
Data for Figure 1, photocurrent properties of ChRs.
- https://cdn.elifesciences.org/articles/83975/elife-83975-fig1-data1-v1.xlsx
Figure 1—figure supplement 1
Light power dependency of photocurrent amplitude of GtCCR4-3.0 (A), KnChR (B), CrChR2[T159C] (C), CoChR (D), and ChrimsonR (E).
Photocurrent amplitude at − 60 mV was plotted as a function of light power. 511 nm light (A), 469 nm light (B, C), and 590 nm light (D) were illuminated. Filled circle: peak photocurrent; open …
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Figure 1—figure supplement 1—source data 1
Data for Figure 1—figure supplement 1, light power dependencies of photo current amplitude of ChRs.
- https://cdn.elifesciences.org/articles/83975/elife-83975-fig1-figsupp1-data1-v1.xlsx
Figure 2 with 6 supplements
Optogenetic activation of hindbrain reticulospinal V2a neurons and cardiomyocytes by channelrhodopsins.
(A) Schematic of experimental devices. A larva was embedded in agarose. The hindbrain region or the heart were irradiated with light. Tail (caudal fin) movements and heartbeats were monitored by a …
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Figure 2—source data 1
Data for Figure 2D, tail movements of Tg larvae expressing ChRs.
- https://cdn.elifesciences.org/articles/83975/elife-83975-fig2-data1-v1.xlsx
Figure 2—video 1
Tail movements in a larva expressing CrChR2[T159C]-mCherry in reticulospinal V2a neurons.
The hindbrain in a 3-dpf Tg(vsx2:GAL4FF); Tg(UAS-hsp70l:CrChR2[T159C]-mCherry, myl7:mCherry) larva was stimulated with 470 nm light for 100 ms. The timing of light stimulation is indicated by a blue …
Figure 2—video 2
Tail movements in a larva expressing GtCCR4-3.0-EYFP in reticulospinal V2a neurons.
The hindbrain in a 3-dpf Tg(vsx2:GAL4FF); Tg(UAS-hsp70l:GtCCR4-3.0-EYFP, myl7:mCherry) larva was stimulated with 520 nm light for 100 ms. The timing of light stimulation is indicated by a green …
Figure 2—video 3
Tail movements in a larva expressing GtCCR4-MT-P2A-TagCFP in reticulospinal V2a neurons.
The hindbrain in a 3-dpf Tg(vsx2:GAL4FF);Tg(UAS-hsp70l:GtCCR4-MT-P2A-TagCFP, myl7:mCherry) larva was stimulated with 520 nm light for 100 ms. The timing of light stimulation is indicated by a green …
Figure 2—video 4
Tail movements in a larva expressing KnChR-3.0-EYFP in reticulospinal V2a neurons.
The hindbrain in a 3-dpf Tg(vsx2:GAL4FF);Tg(UAS-hsp70l:KnChR-3.0-EYFP, myl7:mCherry) larva was stimulated with 470 nm light for 100 ms. The timing of light stimulation is indicated by a blue circle.
Figure 2—video 5
Tail movements in a larva expressing CoChR-tdTomato in reticulospinal V2a neurons.
The hindbrain in a 3-dpf Tg(vsx2:GAL4FF);Tg(UAS:CoChR-tdTomato), larva was stimulated with 470 nm light for 100 ms. The timing of light stimulation is indicated by a blue circle.
Figure 2—video 6
Tail movements in a larva expressing ChrimsonR-tdTomato in reticulospinal V2a neurons.
The hindbrain in a 3-dpf Tg(vsx2:GAL4FF);Tg(UAS-hsp70l:ChrimsonR-tdTomato, myl7:mCherry) larva was stimulated with 520 nm light for 100 ms. The timing of light stimulation is indicated by a green …
Figure 3 with 3 supplements
Optogenetic activation of hindbrain reticulospinal V2a neurons by GtCCR4, KnChR, and CrChR2[T159C].
(A) Light stimulation-dependent locomotion rates of 3-dpf Tg larvae expressing GtCCR4-3.0-EYFP, GtCCR4-MT, KnChR-3.0-EYFP, CrChR2[T159C]-mCherry. The hindbrain area was irradiated with light (0.4 …
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Figure 3—source data 1
Data for Figure 3, optogenetic activation of hindbrain reticulospinal V2a neurons by ChRs.
- https://cdn.elifesciences.org/articles/83975/elife-83975-fig3-data1-v1.xlsx
Figure 3—figure supplement 1
Latency of locomotion in ChR-expressing and non-expressing larvae.
The hindbrain of 3-dpf larvae expressing GtCCR4-3.0-EYFP, GtCCR4-MT, or KnChR-3.0-EYFP, and non-expressing sibling control larvae was irradiated with light (520 nm for GtCCR4-3.0-EYFP, GtCCR4-MT and …
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Figure 3—figure supplement 1—source data 1
Data for Figure 3—figure supplement 1, latency of locomotion in ChR-expressing and non-expressing larvae.
- https://cdn.elifesciences.org/articles/83975/elife-83975-fig3-figsupp1-data1-v1.xlsx
Figure 3—figure supplement 2
Latency and duration of locomotion induced with channelrhodopsins (ChRs) by light of various light intensities.
Latency (A) and duration (B) of evoked tail movements. The hindbrain area of 3-dpf Tg larvae expressing GtCCR4-3.0, KnChR-3.0, and CrChR2[T159C] and control larvae (non-expressing sibling larvae) …
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Figure 3—figure supplement 2—source data 1
Data for Figure 3—figure supplement 2, latency and duration of locomotion induced with ChRs by light of various intensities.
- https://cdn.elifesciences.org/articles/83975/elife-83975-fig3-figsupp2-data1-v1.xlsx
Figure 3—figure supplement 3
Optogenetic activation of hindbrain reticulospinal V2a neurons by CoChR and ChrimsonR.
(A) Light stimulation-dependent locomotion rates of 3-dpf Tg larvae expressing CoChR-tdTomato and ChrimsonR-tdTomato. The hindbrain area was irradiated with light (0.4 mW/mm2) with a wavelength of …
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Figure 3—figure supplement 3—source data 1
Data for Figure 3—figure supplement 3, optogenetic activation of V2a neurons by CoChR and ChrimsonR.
- https://cdn.elifesciences.org/articles/83975/elife-83975-fig3-figsupp3-data1-v1.xlsx
Figure 4 with 6 supplements
Cardiac arrest induced with GtCCR4-3.0, KnChR, CrChR2[T159C], and GtACR1.
(A) Expression of GtCCR4-3.0-EYFP, KnChR-3.0-EYFP, and GtACR1-EYFP in cardiomyocytes of Tg(myl7:GAL4FF);Tg(UAS-hsp70l:GtCCR4-3.0-EYFP, KnChR-3.0-EYFP, or GtACR1-EYFP, myl7:mCherry) larvae. 4-dpf Tg …
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Figure 4—source data 1
Data for Figure 4, cardiac arrest induced with ChRs.
- https://cdn.elifesciences.org/articles/83975/elife-83975-fig4-data1-v1.xlsx
Figure 4—video 1
Heart movements in a larva expressing GtCCR4-3.0-EYFP in cardiomyocytes.
The heart area of Tg(myl7:GAL4FF);Tg(UAS-hsp70l:GtCCR4-3.0-EYFP, myl7:mCherry) was stimulated with 520 nm light for 100 ms. The timing of light stimulation is indicated by a green circle.
Figure 4—video 2
Heart movements in a larva expressing KnChR-3.0-EYFP in cardiomyocytes.
The heart area of Tg(myl7:GAL4FF);Tg(UAS-hsp70l:KnChR-3.0-EYFP, myl7:mCherry) was stimulated with 470 nm light for 100 ms. The timing of light stimulation is indicated by a blue circle.
Figure 4—video 3
Heart movements in a larva expressing CrChR2[T159C]-mCherry in cardiomyocytes.
The heart area of Tg(myl7:GAL4FF);Tg(UAS-hsp70l:CrChR2[T159C]-mCherry, myl7:mCherry) was stimulated with 470 nm light for 100 ms. The timing of light stimulation is indicated by a blue circle.
Figure 4—video 4
Heart movements in a larva expressing GtACR1-EYFP in cardiomyocytes.
The heart area of Tg(myl7:GAL4FF);Tg(UAS-hsp70l:GtACR1-EYFP, myl7:mCherry) was stimulated with 520 nm light for 100 ms. The timing of light stimulation is indicated by a green circle.
Figure 4—video 5
Heart movements in a larva expressing CoChR-tdTomato in cardiomyocytes.
The heart area of Tg(myl7:GAL4FF);Tg(UAS:CoChR-tdTomato) was stimulated with 470 nm light for 100 ms. The timing of light stimulation is indicated by a blue circle.
Figure 4—video 6
Heart movements in a larva expressing ChrimsonR-tdTomato in cardiomyocytes.
The heart area of Tg(myl7:GAL4FF);Tg(UAS-hsp70l:ChrimsonR-tdTomato, myl7:mCherry) was stimulated with 520 nm light for 100 ms. The timing of light stimulation is indicated by a green circle.
Figure 5 with 4 supplements
Cardiac arrest and resumption of heartbeats with GtCCR4-3.0, KnChR, CrChR2[T159C], and GtACR1.
(A) Cardiac arrest rates of 4-dpf Tg larvae expressing GtCCR4-3.0-EYFP, KnChR-3.0-EYFP, CrChR2[T159C]-mCherry, or GtACR1-EYFP in cardiomyocytes. The heart area was irradiated with appropriate light …
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Figure 5—source data 1
Data for Figure 5, cardiac arrest and resumption of heartbeats with ChRs.
- https://cdn.elifesciences.org/articles/83975/elife-83975-fig5-data1-v1.xlsx
Figure 5—figure supplement 1
Cardiac arrest induced with channelrhodopsins (ChRs) by light of various intensities.
Latency to cardiac arrest (A) and time to resumption of heartbeats (B) after light stimulation with ChRs. The heart area of 4-dpf Tg larvae expressing GtCCR4-3.0-EYFP, KnChR-3.0-EYFP, CrChR2[T159C]-m…
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Figure 5—figure supplement 1—source data 1
Data for Figure 5—figure supplement 1, cardiac arrest induced with ChRs by light of various intensities.
- https://cdn.elifesciences.org/articles/83975/elife-83975-fig5-figsupp1-data1-v1.xlsx
Figure 5—figure supplement 2
Cardiac arrest induced with CoChR and ChrimsonR.
(A) Cardiac arrest rate. The heart area of 4-dpf Tg larvae expressing CoChR-tdTomato or ChrimsonR-tdTomato was irradiated with light (470 nm for CoChR; 520 nm for ChrimsonR) for 100 ms at a strength …
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Figure 5—figure supplement 2—source data 1
Data for Figure 5—figure supplement 2, cardiac arrest induced with CoChR and ChrimsonR.
- https://cdn.elifesciences.org/articles/83975/elife-83975-fig5-figsupp2-data1-v1.xlsx
Figure 5—video 1
Ca2+ imaging in the heart of a larva expressing KnChR-3.0-EYFP and GCaMP6s in cardiomyocytes.
The heart area of Tg(myl7:GAL4FF);Tg(UAS-hsp70l:KnChR-3.0-EYFP, myl7:mCherry);Tg(UAS-hsp70l:GCaMP6s) was stimulated with a fluorescence detection filter (excitation 470–495 nm, emission 510–550 nm). …
Figure 5—video 2
Ca2+ imaging in the heart of a larva expressing GtACR1-EYFP and GCaMP6s in cardiomyocytes.
The heart area of Tg(myl7:GAL4FF);Tg(UAS-hsp70l:GtACR1-EYFP, myl7:mCherry);Tg(UAS-hsp70l:GCaMP6s) was stimulated with a fluorescence detection filter (excitation 470–495 nm, emission 510–550 nm). …
Figure 6 with 1 supplement
Optogenetic activation of hindbrain reticulospinal V2a neurons with BeGC1-EGFP.
(A) Expression of BeGC1-EGFP in the hindbrain reticulospinal V2a neurons. 3-dpf Tg(vsx2:GAL4FF);Tg(UAS:BeGC1-EGFP, myl7:mCherry);Tg(UAS:RFP) larvae were fixed and stained with anti-GFP (EGFP, green) …
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Figure 6—source data 1
Data for Figure 6, optogenetic activation of hindbrain reticulospinal V2a neurons with BeGC1-EGFP.
- https://cdn.elifesciences.org/articles/83975/elife-83975-fig6-data1-v1.xlsx
Figure 6—video 1
Tail movements in a larva expressing BeGC1-EGFP in reticulospinal V2a neurons.
The hindbrain in a 3-dpf Tg(vsx2:GAL4FF);Tg(UAS-hsp70l:BeGC1-EGFP, myl7:mCherry) larva was stimulated with 520 nm light for 500 ms. The timing of light stimulation is indicated by a green circle.
Figure 7 with 5 supplements
Optogenetic activation of reticulospinal V2a neurons with bPAC and OaPAC.
(A) Expression of bPAC and OaPAC in reticulospinal V2a neurons. 3-dpf Tg(vsx2:GAL4FF);Tg(UAS-hsp70l:bPAC-MT-T2A-tDimer, myl7:mCherry);Tg(UAS;RFP) and Tg(vsx2:GAL4FF);Tg(UAS-hsp70l:OaPAC-Flag-P2A-TagC…
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Figure 7—source data 1
Data for Figure 7, optogenetic activation of reticulospinal neurons with PACs.
- https://cdn.elifesciences.org/articles/83975/elife-83975-fig7-data1-v1.xlsx
Figure 7—figure supplement 1
Latency of locomotion in PAC-expressing and non-expressing larvae.
The hindbrain of 3-dpf larvae expressing bPAC or OaPAC, and non-expressing sibling control larvae was irradiated with 470 nm light for 500 ms. Six consecutive stimulation trials were analyzed for …
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Figure 7—figure supplement 1—source data 1
Data for Figure 7—figure supplement 1, latency of locomotion in PAC-expressing and non-expressing larvae.
- https://cdn.elifesciences.org/articles/83975/elife-83975-fig7-figsupp1-data1-v1.xlsx
Figure 7—figure supplement 2
Optogenetic control of the heart by bPAC or OaPAC.
(A) Expression of bPAC-MT or OaPAC-Flag in cardiomyocytes. 4-dpf larvae expressing bPAC-MT or OaPAC-Flag were fixed and stained with anti-Myc or Flag (green), and anti-DsRed (mCherry, magenta) …
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Figure 7—figure supplement 2—source data 1
Data for Figure 7—figure supplement 2, optogenetic control of the heart by bPAC.
- https://cdn.elifesciences.org/articles/83975/elife-83975-fig7-figsupp2-data1-v1.xlsx
Figure 7—video 1
Tail movements in a larva expressing bPAC-MT in reticulospinal V2a neurons.
The hindbrain in a 3-dpf Tg(vsx2:GAL4FF);Tg(UAS-hsp70l:bPAC-MT-T2A-tDimer, myl7:mCherry) larva was stimulated with 470 nm light for 500 ms. The timing of light stimulation is indicated by a blue …
Figure 7—video 2
Tail movements in a larva expressing OaPAC-Flag in reticulospinal V2a neurons.
The hindbrain in a 3-dpf Tg(vsx2:GAL4FF);Tg(UAS-hsp70l:OaPAC-Flag-P2A-TagCFP, myl7:mCherry) larva was stimulated with 470 nm light for 500 ms. The timing of light stimulation is indicated by a blue …
Figure 7—video 3
Heart movements in a larva expressing bPAC-MT in cardiomyocytes.
The heart area of Tg(myl7:GAL4FF);Tg(UAS-hsp70l:bPAC-MT-T2A-tDimer, myl7:mCherry) was stimulated with 470 nm light for 5 s. The timing of light stimulation is indicated by a blue circle.
Tables
Table 1
Optogenetic tools.
Microbial optogenetic tools were expressed in the hindbrain reticulospinal V2a neurons or cardiomyocytes using the Gal4-UAS system.
The expression levels of the tools were determined by immunostaining with anti-tag (MT or Flag) antibodies or anti-fluorescent marker antibodies (anti-GFP and anti-DsRed for EYFP/EGFP and mCherry, respectively) (+, weak; ++, medium; +++, strong expression). The light stimulus-dependent responses (induced swimming or cardiac arrest) are indicated by the percentage of fish that responded to light stimuli. As controls, the responses of sibling larvae that did not express the tools were also examined. ND, not determined.
| Type | Tool name | Origin | Detection | Stimulation light (nm) | V2a neurons | Heart | ||
|---|---|---|---|---|---|---|---|---|
| Expression | Swimming response (control) | Expression | Cardiac response(control) | |||||
| Channelrhodopsin | ||||||||
| Cation | GtCCR4- 3.0-EYFP | Guillardia theta | EYFP fusion | 520 | ++ | 62.5%, n = 8 (14.6%, n = 8) | +++ | 100%, n = 4 (0%, n = 4) |
| Cation | GtCCR4-MT | Guillardia theta | MT-P2A-TagCFP | 520 | +++ | 50.0%, n = 8 (8.33%, n=8) | ND | ND |
| Cation | KnChR-3.0-EYFP | Klebsormidium nitens | EYFP fusion | 470 | ++* | 100%, n = 8 (8.33%, n = 8) | +++ | 100%, n = 4 (0%, n = 4) |
| Cation | CrChR2 [T159C]-mCherry | Chlamydomonas reinhardtii | mCherry fusion | 470 | ++ | 67.4%, n = 12 (25.0%, n = 8) | + ¶ | 100%, n = 4 (0%, n = 4) |
| Cation | CoChR-tdTomato | Chloromonas oogama | tdTomato fusion | 470 | +++† | 100%, n = 8 (2.08%, n = 8) | ND | 100%, n = 4 |
| Cation | ChrimsonR-tdTomato | Chlamydomonas noctigama | tdTomato fusion | 520 | ++† | 23.8%, n = 8 (6.25%, n = 8) | ND | 100%, n = 4 |
| Anion | GtACR1-EYFP | Guillardia theta | EYFP fusion | 520 | ++ ‡ | 80.5%, n = 6 § (13.2%, n = 6) | +++ | 100%, n = 4 (0%, n = 4) |
| Enzymorhodopsin | ||||||||
| Guanyly cyclase | BeGC1-EGFP | Blastocladiella emersonii | EGFP fusion | 520 | +++ | 53.5%, n = 8 (10.4%, n = 8) | +++** | 0%, n = 102†† (ND) |
| Photoactivated adenylyl cyclase | ||||||||
| Adenylyl cyclase | bPAC-MT | Beggiatoa | MT-T2A-tDimer | 470 | ++ | 60.6%, n = 8 (26.2%, n = 8) | +++ | 100%, n = 4 ‡ ‡ (ND) |
| Adenylyl cyclase | OaPAC-Flag | Oscillatoria acuminata | Flag-P2A-TagCFP | 470 | ++ | 42.5%, n = 8 (15.4%, n = 8) | +++ | 0%, n = 104 § § (ND) |
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*
Expression of KnChR-3.0-EYFP was mosaic.
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†
Expression of CoChR-tdTomato and ChrimsonR-tdTomato was detected by observation with an epifluorescent stereomicroscope.
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‡
Expression was confirmed by detecting EYFP.
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§
The percentages of spontaneous tail movements elicited by white light that was inhibited by rhodopsin activation (locomotion-inhibition trials) are indicated (no rhodopsin activation was used as the control).
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¶
Expression of CrChR2 [T159C]-mCherry was detected by qPCR.
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**
The expression of BeGC1-EGFP was determined by observation with an epifluorescent microscope MZ16 FA and a fluorescence detection filter (460–500 nm, Leica).
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††
Cardiac arrest was not induced with 490–510 nm, 530–560 nm (epifluorescent stereomicroscope-equipped light source, n = 100), or 520 nm (LED) light stimuli (n = 2).
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‡ ‡
Light stimulation with 470 nm LED light for 5 s induced bradycardia, which took a few minutes to return to normal heartbeats.
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§ §
Stimulation with 460–500 nm (epifluorescent stereomicroscope-equipped light source, n = 100) or 470 nm LED light (n = 4) induced neither cardiac arrest nor bradycardia, while stimulation with 470 nm LED light occasionally induced transient tachycardia for a few seconds (n = 2).
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Gene (Guillardia theta) | GtCCR4 | Yamauchi et al., 2017 | GenBank: MF039475.1 | |
| Gene (Klebsormidium nitens) | KnChR | Tashiro et al., 2021 | GenBank: DF236986.1, GAQ79757.1 | |
| Gene (Chlamydomonas reinhardtii) | CrChR2[T159C] | Berndt et al., 2011 | ||
| Gene (Chloromonas oogama) | CoChR | Klapoetke et al., 2014 | ||
| Gene (Chlamydomonas noctigama) | ChrimsonR | Klapoetke et al., 2014 | ||
| Gene (Blastocladiella emersonii) | BeGC1 | Scheib et al., 2015 | GenBank: KP731361.1 | |
| Gene (G. theta) | GtACR1 | Govorunova et al., 2015 | GenBank: KP171708.1 | |
| Gene (Beggiatoa sp.) | bPAC | Stierl et al., 2011 | GenBank: GU461306.2 | |
| Gene (Oscillatoria acuminata) | OaPAC | Ohki et al., 2016 | GenPept: WP_015149803.1 | |
| Genetic reagent (Danio rerio) | mitfaw2/w2 | Lister et al., 1999 | ||
| Genetic reagent (D. rerio) | TgBAC(vsx2:GAL4FF) | Kimura et al., 2013 | TgBAC(vsx2:GAL4FF) nns18Tg | |
| Genetic reagent (D. rerio) | Tg(myl7:GAL4FF) | Accompanying paper | Tg(myl7:GAL4FF)nub38Tg | Available from M. Hibi lab |
| Genetic reagent (D. rerio) | Tg(UAS-hsp70l:GtCCR4-3.0-EYFP) | This paper | Tg(5xUAS-hsp70l:GtCCR4- 3.0-EYFP, myl7:mCherry)nub49Tg | Available from M. Hibi lab |
| Genetic reagent (D. rerio) | Tg(UAS:CoChR-tdTomato) | This paper | Tg(14xUAS-E1b:CoChR-tdTomato)nub120Tg | Available from M. Hibi lab |
| Genetic reagent (D. rerio) | Tg(UAS-hsp70l:ChrimsonR-tdTomato) | This paper | Tg(5xUAS-hsp70l:ChrimsonR, myl7:mCherry)nub119Tg | Available from M. Hibi lab |
| Genetic reagent (D. rerio) | Tg(UAS-hsp70l:GtCCR4-MT-P2A-TagCFP) | This paper | Tg(5xUAS-hsp70l:GtCCR4- MT-P2A-TagCFP, myl7:mCherry)nub50Tg | Available from M. Hibi lab |
| Genetic reagent (D. rerio) | Tg(UAS-hsp70l:KnChR-3.0-EYFP) | This paper | Tg(5xUAS-hsp70l:KnChR-3.0-EYFP, myl7:mCherry)nub51Tg | Available from M. Hibi lab |
| Genetic reagent (D. rerio) | Tg(UAS-hsp70l:CrChR2[T159C]-mCherry) | This paper | Tg(5xUAS-hsp70l:CrChR2 [T159C]-mCherry, myl7:mCherry)nub52Tg | Available from M. Hibi lab |
| Genetic reagent (D. rerio) | Tg(UAS-hsp70l:GtACR1-EYFP) | This paper | Tg(5xUAS-hsp70l:GtACR1- EYFP, myl7:mCherry)nub53Tg | Available from M. Hibi lab |
| Genetic reagent (D. rerio) | Tg(UAS:CoChR-tdTomato) | This paper | Tg(14xUAS-E1b:CoChR-tdTomato)nub120Tg | Available from M. Hibi lab |
| Genetic reagent (D. rerio) | Tg(UAS-hsp70l:ChrimsonR-tdTomato) | This paper | Tg(5xUAS-hsp70l:ChrimsonR-tdTomato)nub119Tg | Available from M. Hibi Lab |
| Genetic reagent (D. rerio) | Tg(UAS-hsp70l:BeGC1-EGFP) | This paper | Tg(5xUAS-hsp70l:BeGC1-EGFP, myl7:mCherry)nub54Tg | Available from M. Hibi lab |
| Genetic reagent (D. rerio) | Tg(UAS-hsp70l:bPAC-MT-T2A-tDimer) | This paper | Tg(5xUAS-hsp70l:bPAC-MT-T2A-tDimer, myl7:mCherry)nub55Tg | Available from M. Hibi lab |
| Genetic reagent (D. rerio) | Tg(UAS-hsp70l:OaPAC-Flag-P2A-TagCFP) | This paper | Tg(5xUAS-hsp70l: OaPAC-Flag-P2A-TagCFP, myl7:mCherry)nub56Tg | Available from M. Hibi lab |
| Genetic reagent (D. rerio) | Tg(UAS-hsp70l:GCaMP6s) | Muto et al., 2017 | Tg(5xUAS-hsp70l:GCaMP6s) nkUAShspzGCaMP6s13aTg | |
| Genetic reagent (D. rerio) | Tg(elavl3:Flamindo2) | This paper | Tg(elavl3:Flamindo2)nub57TG | Available from M. Hibi lab |
| Cell line (hybrid of Rattus norvegicus and Mus musculus) | ND7/23 | Wood et al., 1990 | ECACC 92090903 | https://www.saibou.jp/en/reagents/ |
| Recombinant DNA reagent | pCS2+GtCCR4-3.0-EYFP | This paper | Available from M. Hibi lab | |
| Recombinant DNA reagent | pCS2+GtCCR4- MT-P2A-TagCFP | This paper | Available from M. Hibi lab | |
| Recombinant DNA reagent | pCS2+CrChR2 [T159C]-mCherry | This paper | Available from M. Hibi lab | |
| Recombinant DNA reagent | pBH-R1-R2 | Accompanying paper | Available from M. Hibi lab | |
| Recombinant DNA reagent | pENTR L1-5xUAS-hsp70l-R5 | Accompanying paper | Available from M. Hibi lab | |
| Recombinant DNA reagent | pENTR L5-GtCCR4-MT-P2A-TagCFP-SV40pAS-L2 | This paper | Available from M. Hibi lab | |
| Recombinant DNA reagent | pENTR L5-KnChR-3.0-EYFP-SV40pAS -L2 | This paper | Available from M. Hibi lab | |
| Recombinant DNA reagent | pENTR L5-GtACR1-EYFP-SV40pAS-L2 | This paper | Available from M. Hibi lab | |
| Recombinant DNA reagent | pENTR L5-CrChR2[T159C]-mCherry-SV40pAS-L2 | This paper | Available from M. Hibi lab | |
| Recombinant DNA reagent | pENTR L5-BeGC1-EGFP-SV40pAS-L2 | This paper | Available from M. Hibi lab | |
| Recombinant DNA reagent | pENTR L5-BeGC1-EGFP-SV40pAS-L2 | This paper | Available from M. Hibi lab | |
| Recombinant DNA reagent | pENTR L5-bPAC-MT-T2A-tDimer-SV40pAS-L2 | This paper | Available from M. Hibi lab | |
| Recombinant DNA reagent | pENTR L5-OaPAC-Flag-P2A-TagCFP-SV40pAS-L2 | This paper | Available from M. Hibi lab | |
| Antibody | Mouse monoclonal anti-Flag antibody | Sigma-Aldrich | Cat# F3165; RRID:AB_259529 | Dilution 1:500 |
| Antibody | Mouse monoclonal anti-Myc tag antibody | Santa Cruz Biotechnology | Cat# sc-40; RRID:AB_627268 | Dilution 1:500 |
| Antibody | Rat monoclonal anti-GFP antibody | Nacalai Tesque, Inc. | Cat# 04404-84; RRID:AB_10013361 | Dilution 1:500 |
| Antibody | Rabbit polyclonal anti-DsRed antibody | Takara Bio | Cat# 632496; RRID:AB_10013483 | Dilution 1:500 |
| Antibody | Goat CF488A anti-mouse IgG antibody | Biotium, Inc | Cat# 20018; RRID:AB_10557263 | Dilution 1:500 |
| Antibody | Goat CF488A anti-rat IgG antibody | Biotium, Inc | Cat# 20023; RRID: AB_10557403 | Dilution 1:500 |
| Antibody | Goat CF568 anti-rabbit IgG antibody | Biotium, Inc | Cat# 20103; RRID:AB_10558012 | Dilution 1:500 |
| Chemical compound, drug | tricaine methanesulfonate | Nacalai Tesque, Inc | Cat# 01916-32 | |
| Chemical compound, drug | low gelling temperature Type VII-A | Sigma-Aldrich | A0701 | |
| Chemical compound, drug | pentylenetetrazol | Sigma-Aldrich | Cat# P6500 | |
| Software, algorithm | SutterPatch 1.1.1 | Sutter Instrument Co. | https://www.sutter.com/AMPLIFIERS/SutterPatch.html | |
| Software, algorithm | pCLAMP10.6 | Molecular Devices | https://support.moleculardevices.com/s/article/Axon-pCLAMP-10-Electrophysiology-Data-Acquisition-Analysis-Software-Download-Page | |
| Software, algorithm | PolyScan2 | Mightex | ||
| Software, algorithm | StreamPix7 | NorPix Inc | ||
| Software, algorithm | LabVIEW | National Instruments | 2015 | https://www.ni.com/ja-jp.html |
| Software, algorithm | GraphPad Prism5 | GraphPad Software | https://www.mdf-soft.com/ | |
| Software, algorithm | VSDC Free Video Editor 6.4.7.155 | FLASH-INTEGRO LLC | https://www.videosoftdev.com/jp | |
| Software, algorithm | Microsoft Movies & TV | Microsoft Corp. | https://apps.microsoft.com/store/detail/movies-tv/9WZDNCRFJ3P2 | |
| Software, algorithm | QuickTime player 10.5 | Apple Inc. | https://quicktime.softonic.jp/ | |
| Software, algorithm | Fiji/ImageJ | National Institutes of Health (NIH) | http://fiji.sc/ | |
| Software, algorithm | R 3.6.1 and 4.2.1 | https://www.r-project.org/ | ||
| Software, algorithm | ggplot2 3.2.0 of R | https://ggplot2.tidyverse.org | ||
| Software, algorithm | nlme 3.1–162 of R | https://cran.r-project.org/web/packages/nlme/index.html | ||
| Software, algorithm | Bonsai | Lopes et al., 2015 | https://open-ephys.org/bonsai | |
| Software, algorithm | Python 3.5.6 | Python Software Foundation | https://www.python.org/ | |
| Software, algorithm | Tracker Video Analysis and Modeling Tool for Physics Education 5.1.5 | https://physlets.org/tracker/ | ||
| Software, algorithm | Microsoft Excel for Mac, ver. 16.74 | Microsoft | ||
| Software, algorithm | HB_frequency.py | This paper | Source code file |
Additional files
-
MDAR checklist
- https://cdn.elifesciences.org/articles/83975/elife-83975-mdarchecklist1-v1.pdf
-
Source code 1
Software that detects the timing of each heartbeat from the data of luminosity changes over time.
- https://cdn.elifesciences.org/articles/83975/elife-83975-code1-v1.zip
