Distinct inflammatory and wound healing responses to complex caudal fin injuries of larval zebrafish
- University of Wisconsin-Madison, United States
- Northern Arizona University, United States
Figures
Figure 1 with 1 supplement
Caudal fin recovers from thermal injury, while wound healing is impaired in the presence of infection.
(A) Experimental schematic and analyses. (B) Single-plane brightfield images of caudal fin area of individual wild-type larvae over time in response to simple transection or thermal injury, and in (D…
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Figure 1—source data 1
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Figure 1—source code 1
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Figure 1—figure supplement 1
L. monocytogenes infection during tail wound infection is associated with minimal dissemination and negligible effect on host survival.
(A) Wild-type larvae were wounded (B) or unwounded in presence of mCherry-expressing L. monocytogenes and fixed at indicated time points. 200-micron size z series at 5-micron steps was acquired …
Figure 2 with 3 supplements
Caudal fin injuries trigger distinct inflammatory responses.
(A) Sum-projections of z-stacks acquired by laser scanning confocal microscope using triple transgenic larvae (Tg(tnf:GFP) x Tg(lysC:BFP/mpeg1:mCherry-CAAX)) over time in response to simple …
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Figure 2—figure supplement 1
Leukocyte recruitment in response to thermal injury.
Single channels associated with merged images in Figure 2A showing (A) transection and (B) burn image sets. Neutrophils, macrophages and TNFα were labeled with BFP, mCherry and GFP respectively. …
Figure 2—figure supplement 2
Leukocyte recruitment in the caudal fin tissue is minimal in unwounded larvae.
Additional p values associated with Figure 2B and C are provided for (A) neutrophils and (B) macrophages to reflect changes over time within each injury. Leukocyte recruitment in unwounded larvae or …
Figure 2—figure supplement 3
Leukocyte recruitment in response to transection in the presence of infection.
Single channels associated with merged images in Figure 2D showing (A) uninfected and (B) L. monocytogenes (Lm)- infected image sets. Neutrophils, macrophages and TNFα were labeled with BFP, mCherry …
Figure 3 with 2 supplements
Multiphoton microscopy of collagen and vimentin-expressing cells identify differences in tissue remodeling over time amongst the injuries.
(A) Z-projections of caudal fins imaged at different times after transection, thermal injury (burn) or infected transection showing the tissue wound edge in the bright field and corresponding SHG …
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Figure 3—figure supplement 1
VimGFP-positive cells at the wound recover over time at the Lm-infected tail wound.
(A) an example of an image with vimentin expression showing a 50 × 100 micron box placed strategically to count vimGFP-positive cells is shown (see Materials and methods). (B) Merge of z-projections …
Figure 3—figure supplement 2
Tail wound infection in the presence of hly mutant of L. monocytogenes causes no delay in vimGFP-positive cells and is associated with limited inflammation.
(A) representative images of sum projections of z series acquired by spinning disk confocal microscope using Tg(vim:GFP) larvae following tail transection in the presence of mCherry-expressing …
Figure 4 with 2 supplements
Wound healing and wound-associated vimentin-expressing cells are regulated by Stat3 and TGFβ signaling.
(A) Single-plane brightfield images of wild-type, heterozygous and stat3 mutant larvae showing tissue regrowth of caudal fins at 72 hr post tail transection (hptt) and (B) corresponding …
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Figure 4—source data 1
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Figure 4—figure supplement 1
Stat3 or TGFβ signaling does not affect the presence of vimGFP-positive cells associated with development.
(A) Sum-projections of z-stacks acquired by spinning disk confocal microscope using Tg(vim:GFP) larvae in stat3 mutant background, showing tail fin of age-matched unwounded larvae at 6 dpf. Scale …
Figure 4—figure supplement 2
Table summarizing the characteristics of the three wound models.
https://doi.org/10.7554/eLife.45976.035Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (10403S) | Listeria monocytogenes (strain 10403S) | Vincent et al., 2016 | was be obtained from JD Sauer Lab, University of Wisconsin - Madison | |
| Strain, strain background (D. Rerio) | WT (AB) | ZIRC | ZL1 | https://zebrafish.org/home/guide.php |
| Strain, strain background (D. Rerio) | Tg(tnf:GFP) | Marjoram et al., 2015 | was obtain from M Bagnat Lab, Duke University | |
| Strain, strain background (D. Rerio) | Tg(vim:GFP) | LeBert et al., 2018 | can be obtained from A Huttenlocher Lab, University of Wisconsin - Madison | |
| Strain, strain background (D. Rerio) | Tg(lysC:mCherry-histone2b) | Lam et al., 2014 | can be obtained from A Huttenlocher Lab, University of Wisconsin - Madison | |
| Strain, strain background (D. Rerio) | Tg(mpeg1:mCherry-histone2b) | Vincent et al., 2016 | can be obtained from A Huttenlocher Lab, University of Wisconsin - Madison | |
| Strain, strain background (D. Rerio) | Tg(mpeg1:GFP-histone2b) | this paper | can be obtained from A Huttenlocher Lab, University of Wisconsin - Madison | |
| Strain, strain background (D. Rerio) | Tg(lysC:BFP) | Rosowski et al., 2018 | can be obtained from A Huttenlocher Lab, University of Wisconsin - Madison | |
| Strain, strain background (D. Rerio) | Tg(mpeg1:mCherry-CAAX) | Bojarczuk et al., 2016 | was obtained from LI Zon Lab, Boston Children's Hospital, Dana Farber Cancer Institute | |
| Strain, strain background (D. Rerio) | Tg(lysC:BFP/mpeg1:mCherry-CAAX) | de Oliveira et al., 2019 | can be obtained from A Huttenlocher Lab, University of Wisconsin - Madison | |
| Strain, strain background (D. Rerio) | casper | White et al., 2008 | ZL1714 | https://zebrafish.org/home/guide.php |
| Strain, strain background (D. Rerio) | stat3stl27/+ | Liu et al., 2017b | was obtained from L Solnica-Krezel Lab, Washington University School of Medicine, St. Louis | |
| Recombinant DNA reagent (D. Rerio) | tol2-mpeg1:GFP-histone2b | this paper | can be obtained from A Huttenlocher Lab, University of Wisconsin - Madison | |
| Chemical compound, drug | ALK5 inhibitor (ALK5i) | Selleckchem | item no. SB431542; RRID:SCR_003823 | 25 µM in DMSO; no pretreatment, start treatment upon wounding, refresh daily |
| Software, algorithm | GraphPad Prism | RRID:SCR_002798 | https://www.graphpad.com/scientific-software/prism/ | |
| Software, algorithm | SAS | RRID:SCR_008567 | https://www.sas.com/en_us/home.html | |
| Software, algorithm | Fiji, ImageJ | Schindelin et al., 2012 | RRID:SCR_002285 | https://fiji.sc/ |
| Software, algorithm | CurveAlign | Liu et al., 2017a | https://loci.wisc.edu/software/curvealign | |
| Other | High temperature cautery pen, fine tip | Bovie | AA01 | http://www.boviemedical.com/hightempcauteries/ |
Additional files
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Transparent reporting form
- https://doi.org/10.7554/eLife.45976.043
