Damage-induced basal epithelial cell migration modulates the spatial organization of redox signaling and sensory neuron regeneration
- Department of Medical Microbiology and Immunology, University of Wisconsin-Madison, United States
- Cellular and Molecular Biology Graduate Program, University of Wisconsin-Madison, United States
- Department of Biostatistics and Medical Informatics, University of Wisconsin-Madison, United States
- Department of Pediatrics, University of Wisconsin-Madison, United States
Figures
Figure 1 with 1 supplement
Peripheral sensory axons have impaired regeneration after burn injury.
(A) Schematic of larval zebrafish injury. Gray dashed line denotes area used to measure axon density to the right of the notochord. (B) Confocal max-projected images of sensory axons in uninjured, …
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Figure 1—source data 1
Numerical data for Figure 1C.
- https://cdn.elifesciences.org/articles/94995/elife-94995-fig1-data1-v1.xlsx
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Figure 1—source data 2
Numerical data for Figure 1D.
- https://cdn.elifesciences.org/articles/94995/elife-94995-fig1-data2-v1.xlsx
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Figure 1—source data 3
Numerical data for Figure 1F.
- https://cdn.elifesciences.org/articles/94995/elife-94995-fig1-data3-v1.xlsx
Figure 1—figure supplement 1
Elavl3-GCaMP5 transgenic fish show sensory axon damage.
(A) Confocal max-projected images of axon damage in Tg(Elavl3:GCaMP5) larval zebrafish caudal fins either untreated or 30 min post-treatment with the neurotoxin sodium azide (NaN3, 1.5% final …
Figure 2
The burn wound microenvironment contributes to impaired sensory axon regeneration.
(A) Schematic of two-wound experiment design. (B) Confocal max-projected images of FM dye staining following secondary transection in the two-wound experiment at 5 min post-wound (mpw) and 6 hr …
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Figure 2—source data 1
Numerical data for Figure 2D.
- https://cdn.elifesciences.org/articles/94995/elife-94995-fig2-data1-v1.xlsx
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Figure 2—source data 2
Numerical data for Figure 2E.
- https://cdn.elifesciences.org/articles/94995/elife-94995-fig2-data2-v1.xlsx
Figure 3 with 2 supplements
Burn injury induces coordinated keratinocyte and sensory axon movement.
(A) Confocal max-projected time-series images of Tg(Krt4:UtrCH-GFP) larvae after either transection or burn injury. Yellow pseudocolored cells and colored tracks highlight keratinocyte displacement. …
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Figure 3—source data 1
Numerical data for Figure 3A.
- https://cdn.elifesciences.org/articles/94995/elife-94995-fig3-data1-v1.xlsx
Figure 3—figure supplement 1
Basal keratinocyte migration in response to injury.
(A) Confocal time series of basal keratinocyte, Tg(Krtt1c19e:acGFP), movement after the indicated injury. Yellow pseudocolored cells highlight keratinocyte displacement. Scale bars = 20 µm.
Figure 3—figure supplement 2
Sensory axon cell bodies are not displaced following burn injury.
(A) Schematic of Rohon-Beard (RB) (green) and dorsal root ganglia (DRG) (blue) soma localization in 3 days post-fertilization (dpf) zebrafish. Red box denotes area in which the image shown in B was …
Figure 4 with 1 supplement
The Arp 2/3 inhibitor CK666 impairs early keratinocyte movement and alters the spatial distribution of reactive oxygen species signaling.
(A) Confocal max-projected images of control or CK666-treated transiently injected Tg(Krtt1c19e:Lifeact-mRuby) larvae. Arrows point to lamellipodia in the control larva, and lack of lamellipodia in …
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Figure 4—source data 1
Numerical data for Figure 4B.
- https://cdn.elifesciences.org/articles/94995/elife-94995-fig4-data1-v1.xlsx
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Figure 4—source data 2
Numerical data for Figure 4C.
- https://cdn.elifesciences.org/articles/94995/elife-94995-fig4-data2-v1.xlsx
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Figure 4—source data 3
Numerical data for Figure 4E.
- https://cdn.elifesciences.org/articles/94995/elife-94995-fig4-data3-v1.xlsx
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Figure 4—source data 4
Numerical data for Figure 4G.
- https://cdn.elifesciences.org/articles/94995/elife-94995-fig4-data4-v1.xlsx
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Figure 4—source data 5
Numerical data for Figure 4H.
- https://cdn.elifesciences.org/articles/94995/elife-94995-fig4-data5-v1.xlsx
Figure 4—figure supplement 1
Early reactive oxygen species (ROS) inhibition is not sufficient to improve axon regeneration.
(A) Confocal max-projected images of sensory axons treated with diphenyleneiodonium (DPI). (B) Quantification of axon density 24 hr post-wound (hpw). N>19 larvae per condition from three replicates. …
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Figure 4—figure supplement 1—source data 1
Numerical data for Figure 4—figure supplement 1B.
- https://cdn.elifesciences.org/articles/94995/elife-94995-fig4-figsupp1-data1-v1.xlsx
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Figure 4—figure supplement 1—source data 2
Numerical data for Figure 4—figure supplement 1C.
- https://cdn.elifesciences.org/articles/94995/elife-94995-fig4-figsupp1-data2-v1.xlsx
Figure 5
Treatment with isotonic solution inhibits keratinocyte migration and dampens reactive oxygen species (ROS) signaling.
(A) Confocal time-series images of basal keratinocyte movement in Tg(Krtt1c19e:acGFP) larvae over 1 hr post-wound (hpw) after burn injury in the indicated treatment. (B) Plot of basal keratinocyte …
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Figure 5—source data 1
Numerical data for Figure 5B.
- https://cdn.elifesciences.org/articles/94995/elife-94995-fig5-data1-v1.xlsx
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Figure 5—source data 2
Numerical data for Figure 5C.
- https://cdn.elifesciences.org/articles/94995/elife-94995-fig5-data2-v1.xlsx
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Figure 5—source data 3
Numerical data for Figure 5E.
- https://cdn.elifesciences.org/articles/94995/elife-94995-fig5-data3-v1.xlsx
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Figure 5—source data 4
Numerical data for Figure 5G.
- https://cdn.elifesciences.org/articles/94995/elife-94995-fig5-data4-v1.xlsx
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Figure 5—source data 5
Numerical data for Figure 5H.
- https://cdn.elifesciences.org/articles/94995/elife-94995-fig5-data5-v1.xlsx
Figure 6 with 1 supplement
Isotonic treatment improves axon regeneration.
(A) Confocal max-projected images of axon damage in control or isotonic-treated Tg(Elavl3:GCaMP5) larvae 0 or 6 hr post-wound (hpw). (B) Quantification of axon damage in control and isotonic-treated …
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Figure 6—source data 1
Numerical data for Figure 6B.
- https://cdn.elifesciences.org/articles/94995/elife-94995-fig6-data1-v1.xlsx
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Figure 6—source data 2
Numerical data for Figure 6D.
- https://cdn.elifesciences.org/articles/94995/elife-94995-fig6-data2-v1.xlsx
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Figure 6—source data 3
Numerical data for Figure 6E.
- https://cdn.elifesciences.org/articles/94995/elife-94995-fig6-data3-v1.xlsx
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Figure 6—source data 4
Numerical data for Figure 6H.
- https://cdn.elifesciences.org/articles/94995/elife-94995-fig6-data4-v1.xlsx
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Figure 6—source data 5
Numerical data for Figure 6J.
- https://cdn.elifesciences.org/articles/94995/elife-94995-fig6-data5-v1.xlsx
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Figure 6—source data 6
Numerical data for Figure 6K.
- https://cdn.elifesciences.org/articles/94995/elife-94995-fig6-data6-v1.xlsx
Figure 6—figure supplement 1
Keratinocyte movement after injury and effect of D-Sorbitol on sensory axon regeneration.
(A) Representative max-projected confocal images of sensory axons in control and isotonic D-Sorbitol-treated larvae 24 hr post-wound (hpw). (B) Quantification of axon density in wounded tissue 24 …
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Figure 6—figure supplement 1—source data 1
Numerical data for Figure 6—figure supplement 1B.
- https://cdn.elifesciences.org/articles/94995/elife-94995-fig6-figsupp1-data1-v1.xlsx
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Figure 6—figure supplement 1—source data 2
Numerical data for Figure 6—figure supplement 1C.
- https://cdn.elifesciences.org/articles/94995/elife-94995-fig6-figsupp1-data2-v1.xlsx
Appendix 1—figure 1
Drug treatments do not affect axon density in unwounded larvae.
(A) Representative max-projected confocal images of sensory axons in unwounded, drug-treated 4 dpf larvae. (B) Quantification of axon density in larvae treated as stated in E. N>15 larvae each from …
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Appendix 1—figure 1—source data 1
Numerical data for Appendix 1—figure 1.
- https://cdn.elifesciences.org/articles/94995/elife-94995-app1-fig1-data1-v1.xlsx
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Appendix 1—figure 1—source data 2
Numerical data for Appendix 1—figure 1.
- https://cdn.elifesciences.org/articles/94995/elife-94995-app1-fig1-data2-v1.xlsx
Author response image 1
Author response image 2
Videos
Video 1
Burn injury induces keratinocyte movement.
Tg(Krt4:UtrCH-GFP) larvae were injured either by tailfin transection (left) or burn (right). While minimal keratinocyte movement is observed following transection, burn injury results in …
Video 2
Sensory axons move with associated keratinocytes following injury.
Basal keratinocyte (magenta), Tg(Krtt1c19e:Lifeact-Ruby), and sensory axon (cyan), Tg(Ngn1:GFP-Caax), movement was tracked following burn injury. Arrows highlight regions where keratinocyte and …
Video 3
Tissue movement is associated with axonal damage following burn injury.
Tg(Elavl3:GCaMP5) larva was burn wounded to track axonal damage, indicated by elevated intracellular calcium (black dots). Damage present at time 0 min is due to the burn wound itself, while new …
Video 4
Wounding in isotonic medium prevents burn-induced keratinocyte movement.
Basal keratinocyte, Tg(Krtt1c19e:acGFP), movement was tracked in control (left) and isotonic-treated (right) larvae following burn injury. Wounding in the presence of isotonic medium prevents …
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain background (D. rerio) | WT (AB) | ZIRC | ZL1 | https://zebrafish.org/home/guide.php |
| Strain background (D. rerio) | Tg(Ngn1:GFP-Caax) | Blader et al., 2003 | ||
| Strain background (D. rerio) | Tg(Krt4:LifeAct-mRuby) | Lam et al., 2015 | ||
| Strain background (D. rerio) | Tg(Krt4:UtrCH-GFP) | Lam et al., 2015 | ||
| Strain background (D. rerio) | Tg(Krt4:TdTomato) | Huttenlocher lab | ||
| Strain background (D. rerio) | TgBac(Lamc1:Lamc1-sfGFP) | Yamaguchi et al., 2022 | ||
| Strain background (D. rerio) | Tg(ElavI3:GCaMP5) | Akerboom et al., 2012 | Received from Jan Huisken lab | |
| Strain background (D. rerio) | Tg(Krtt1c19e:LifeAct-mRuby) | This paper | ||
| Strain background (D. rerio) | Tg(Krtt1c19e:acGFP) | Lee et al., 2014 | Received from Alvaro Sagasti lab | |
| Chemical compound, drug | FM 1-43 dye | Life Technologies | ||
| Chemical compound, drug | NaCl | Fisher Scientific | CAS 7647-14-5 | |
| Chemical compound, drug | D-Sorbitol | Sigma-Aldrich | CAS 50-70-4 | |
| Chemical compound, drug | CK666 (Arp2/3 inhibitor) | Sigma-Aldrich | CAS 442633-00-3 | |
| Chemical compound, drug | Sodium Azide | Fisher Scientific | CAS 26628-22-8 | |
| Chemical compound, drug | Pentafluorobenzenesulfonyl fluorescein | Santa Cruz | CAS 728912-45-6 | |
| Other | Cautery pen, fine tip | Bovie | AA01 | https://www.delasco.com/geiger/ |
| Other | Surgical blade No. 10 | Feather | 2976 | |
| Software, algorithm | GraphPad Prism | RRID:SCR_002798 | https://www.graphpad.com/scientific-software/prism/ | |
| Software, algorithm | Fuji, ImageJ | Schneider et al., 2012 | RRID:SCR_002285 | https://fiji.sc/ |
Additional files
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MDAR checklist
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Source data 1
Primer source data.
- https://cdn.elifesciences.org/articles/94995/elife-94995-data1-v1.xlsx
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Appendix 1—figure 1—source data 1
Numerical data for Appendix 1—figure 1.
- https://cdn.elifesciences.org/articles/94995/elife-94995-app1-fig1-data1-v1.xlsx
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Appendix 1—figure 1—source data 2
Numerical data for Appendix 1—figure 1.
- https://cdn.elifesciences.org/articles/94995/elife-94995-app1-fig1-data2-v1.xlsx
