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Zebrafish fin regeneration after cryoinjury-induced tissue damage.

Chassot B, Pury D, Jaźwińska A - Biol Open (2016)

Bottom Line: In contrast to the common transection model, the damaged part of the fin was spontaneously shed within two days after cryoinjury.Between two and seven days after cryoinjury, this reparative/proliferative phase was morphologically featured by displaced fragments of broken bones.Live imaging of epithelial and osteoblastic transgenic reporter lines revealed that the tissue-specific regenerative programmes were initiated after the clearance of damaged material.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, University of Fribourg, Chemin du Musée 10, Fribourg 1700, Switzerland.

No MeSH data available.


Related in: MedlinePlus

Dynamics of bone and actinotrichia regeneration in cryoinjured fins. (A-E′) Live-imaging of shh:GFP transgenic fish demarcates a subset of cells in the basal layer of the lateral wound epidermis (green) at 2 dpa (A) different time points after cryoinjury (B-E). Dashed line indicates the plane of amputation. The expression of shh:GFP becomes detectable starting at 7 dpci (D,D′), indicating organization and subdivision of the basal epithelium. N=4. Boxes in A-E magnified in A′-E′. (F-J′) Live-imaging of transgenic fish osterix(sp7):GFP highlights intermediately differentiated osteoblasts (green) at 2 dpa (F) and at different time points after cryoinjury (G-J). The expression of osterix(sp7):GFP becomes detectable starting at 7 dpci (I,I′), indicating bone regeneration. N=4. Boxes in F-J magnified in F′-J′. (K-N) Bright-field (upper panels) and confocal (lower panels) images of whole-mount fins immunostained with anti-And1 antibodies (green) at 3 dpa (K) and at different times after cryoinjury (L-N). Bone matrix is detected by autofluorescence (blue). The bright-field images show dark necrotic tissue at the margin of cryoinjured fins. The expression of And1 starts at 5 dpci (M) and becomes more evident at 7 dpci (N). N=4. Yellow dashed line indicates the amputation plane. Scale bar in A=1 mm and in K=100 µm.
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BIO016865F8: Dynamics of bone and actinotrichia regeneration in cryoinjured fins. (A-E′) Live-imaging of shh:GFP transgenic fish demarcates a subset of cells in the basal layer of the lateral wound epidermis (green) at 2 dpa (A) different time points after cryoinjury (B-E). Dashed line indicates the plane of amputation. The expression of shh:GFP becomes detectable starting at 7 dpci (D,D′), indicating organization and subdivision of the basal epithelium. N=4. Boxes in A-E magnified in A′-E′. (F-J′) Live-imaging of transgenic fish osterix(sp7):GFP highlights intermediately differentiated osteoblasts (green) at 2 dpa (F) and at different time points after cryoinjury (G-J). The expression of osterix(sp7):GFP becomes detectable starting at 7 dpci (I,I′), indicating bone regeneration. N=4. Boxes in F-J magnified in F′-J′. (K-N) Bright-field (upper panels) and confocal (lower panels) images of whole-mount fins immunostained with anti-And1 antibodies (green) at 3 dpa (K) and at different times after cryoinjury (L-N). Bone matrix is detected by autofluorescence (blue). The bright-field images show dark necrotic tissue at the margin of cryoinjured fins. The expression of And1 starts at 5 dpci (M) and becomes more evident at 7 dpci (N). N=4. Yellow dashed line indicates the amputation plane. Scale bar in A=1 mm and in K=100 µm.

Mentions: The epithelial-mesenchymal interactions are fundamental to the execution of developmental and regenerative programmes after fin amputation (Blum and Begemann, 2012; Gemberling et al., 2013; Yoshinari and Kawakami, 2011). Sonic hedgehog (Shh) is one of the factors produced by the lateral basal wound epithelium of the outgrowth, suggested to be involved in regeneration of the underlying bones (Blum and Begemann, 2015; Laforest et al., 1998; Quint et al., 2002). To analyse the expression of shh after cryoinjury, we performed time-lapse imaging of shh:GFP transgenic fish. In the fin amputation model, shh:GFP has been shown to be induced at 2 dpa in the wound epithelium of the regenerating rays (Fig. 8A,A′) (Zhang et al., 2012). In contrast, the expression of the transgene was initiated only at 7 dpci in our injury method (Fig. 8B-D′). A robust expression of this transgenic reporter was observed at 9 dpci, indicating a substantial delay in the reactivation of the regenerative programme related to shh gene (Fig. 8E,E′). We concluded that the wound epithelial subdomains become organized after the completion of the reparative phase in the stump.Fig. 8.


Zebrafish fin regeneration after cryoinjury-induced tissue damage.

Chassot B, Pury D, Jaźwińska A - Biol Open (2016)

Dynamics of bone and actinotrichia regeneration in cryoinjured fins. (A-E′) Live-imaging of shh:GFP transgenic fish demarcates a subset of cells in the basal layer of the lateral wound epidermis (green) at 2 dpa (A) different time points after cryoinjury (B-E). Dashed line indicates the plane of amputation. The expression of shh:GFP becomes detectable starting at 7 dpci (D,D′), indicating organization and subdivision of the basal epithelium. N=4. Boxes in A-E magnified in A′-E′. (F-J′) Live-imaging of transgenic fish osterix(sp7):GFP highlights intermediately differentiated osteoblasts (green) at 2 dpa (F) and at different time points after cryoinjury (G-J). The expression of osterix(sp7):GFP becomes detectable starting at 7 dpci (I,I′), indicating bone regeneration. N=4. Boxes in F-J magnified in F′-J′. (K-N) Bright-field (upper panels) and confocal (lower panels) images of whole-mount fins immunostained with anti-And1 antibodies (green) at 3 dpa (K) and at different times after cryoinjury (L-N). Bone matrix is detected by autofluorescence (blue). The bright-field images show dark necrotic tissue at the margin of cryoinjured fins. The expression of And1 starts at 5 dpci (M) and becomes more evident at 7 dpci (N). N=4. Yellow dashed line indicates the amputation plane. Scale bar in A=1 mm and in K=100 µm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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BIO016865F8: Dynamics of bone and actinotrichia regeneration in cryoinjured fins. (A-E′) Live-imaging of shh:GFP transgenic fish demarcates a subset of cells in the basal layer of the lateral wound epidermis (green) at 2 dpa (A) different time points after cryoinjury (B-E). Dashed line indicates the plane of amputation. The expression of shh:GFP becomes detectable starting at 7 dpci (D,D′), indicating organization and subdivision of the basal epithelium. N=4. Boxes in A-E magnified in A′-E′. (F-J′) Live-imaging of transgenic fish osterix(sp7):GFP highlights intermediately differentiated osteoblasts (green) at 2 dpa (F) and at different time points after cryoinjury (G-J). The expression of osterix(sp7):GFP becomes detectable starting at 7 dpci (I,I′), indicating bone regeneration. N=4. Boxes in F-J magnified in F′-J′. (K-N) Bright-field (upper panels) and confocal (lower panels) images of whole-mount fins immunostained with anti-And1 antibodies (green) at 3 dpa (K) and at different times after cryoinjury (L-N). Bone matrix is detected by autofluorescence (blue). The bright-field images show dark necrotic tissue at the margin of cryoinjured fins. The expression of And1 starts at 5 dpci (M) and becomes more evident at 7 dpci (N). N=4. Yellow dashed line indicates the amputation plane. Scale bar in A=1 mm and in K=100 µm.
Mentions: The epithelial-mesenchymal interactions are fundamental to the execution of developmental and regenerative programmes after fin amputation (Blum and Begemann, 2012; Gemberling et al., 2013; Yoshinari and Kawakami, 2011). Sonic hedgehog (Shh) is one of the factors produced by the lateral basal wound epithelium of the outgrowth, suggested to be involved in regeneration of the underlying bones (Blum and Begemann, 2015; Laforest et al., 1998; Quint et al., 2002). To analyse the expression of shh after cryoinjury, we performed time-lapse imaging of shh:GFP transgenic fish. In the fin amputation model, shh:GFP has been shown to be induced at 2 dpa in the wound epithelium of the regenerating rays (Fig. 8A,A′) (Zhang et al., 2012). In contrast, the expression of the transgene was initiated only at 7 dpci in our injury method (Fig. 8B-D′). A robust expression of this transgenic reporter was observed at 9 dpci, indicating a substantial delay in the reactivation of the regenerative programme related to shh gene (Fig. 8E,E′). We concluded that the wound epithelial subdomains become organized after the completion of the reparative phase in the stump.Fig. 8.

Bottom Line: In contrast to the common transection model, the damaged part of the fin was spontaneously shed within two days after cryoinjury.Between two and seven days after cryoinjury, this reparative/proliferative phase was morphologically featured by displaced fragments of broken bones.Live imaging of epithelial and osteoblastic transgenic reporter lines revealed that the tissue-specific regenerative programmes were initiated after the clearance of damaged material.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, University of Fribourg, Chemin du Musée 10, Fribourg 1700, Switzerland.

No MeSH data available.


Related in: MedlinePlus