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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

Detachment of the destroyed fin tissue is associated with displacement and resorption of the dead bone fragments at the wound margin. (A,B) Imaging of bones in the same fin detected by autofluorescence of the mineralized matrix at 2 and 6 dpci. The margin of the remnant fins contains detached and displaced bone fragments between the rays that become resolved (arrows). N=5. (C-G) Confocal imaging of whole-mount fins immunostained with the osteoblast marker Zns5 (red), phagocyte marker L-plastin (green) and autofluorescent bone matrix (blue) at 2 dpa (C) and at different time points after cryoinjury (D-G). At 2 dpa (C), Zns5-labelled osteoblasts accumulate at the tip of the bone to initiate bone regeneration. L-plastin-expressing cells are present in the entire tissue. At 2 dpci (D) and 3 dpci (E), osteoblasts are scattered along the bones in irregular manner. At 5 dpci (F), Zns5-positive cells are enriched at the tips of the intact bones, below the margin of the stump that contains bone debris devoid of osteoblasts. At 7 dpci (G), Zns5 immunostaining is robustly enhanced along the remaining bones, indicating resumed regeneration. L-plastin-expressing cells are associated with the repairing and regenerating tissue. N=4. Scale bar in A=200 µm, in C=100 µm.
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BIO016865F5: Detachment of the destroyed fin tissue is associated with displacement and resorption of the dead bone fragments at the wound margin. (A,B) Imaging of bones in the same fin detected by autofluorescence of the mineralized matrix at 2 and 6 dpci. The margin of the remnant fins contains detached and displaced bone fragments between the rays that become resolved (arrows). N=5. (C-G) Confocal imaging of whole-mount fins immunostained with the osteoblast marker Zns5 (red), phagocyte marker L-plastin (green) and autofluorescent bone matrix (blue) at 2 dpa (C) and at different time points after cryoinjury (D-G). At 2 dpa (C), Zns5-labelled osteoblasts accumulate at the tip of the bone to initiate bone regeneration. L-plastin-expressing cells are present in the entire tissue. At 2 dpci (D) and 3 dpci (E), osteoblasts are scattered along the bones in irregular manner. At 5 dpci (F), Zns5-positive cells are enriched at the tips of the intact bones, below the margin of the stump that contains bone debris devoid of osteoblasts. At 7 dpci (G), Zns5 immunostaining is robustly enhanced along the remaining bones, indicating resumed regeneration. L-plastin-expressing cells are associated with the repairing and regenerating tissue. N=4. Scale bar in A=200 µm, in C=100 µm.

Mentions: To characterize the defects in cryoinjured fins, we analysed the morphology of the remaining bones after tissue sloughing. Live imaging of fins at 2 dpci revealed an accumulation of bone fragments at the margin of the stump (Fig. 5A). The tips of the bones were released from the rays and often displaced perpendicularly to the original position. Imaging of the same fins after the next 4 days (at 6 dpci) revealed that the remnants of mineralized matrix were undergoing degradation and resorption (Fig. 5B). To test whether the detached bone segments were associated with bone-producing cells, the osteoblasts, we performed immunofluorescence staining with Zns5 antibody and we labelled phagocytic cells with anti-L-plastin antibody of whole-mount fins combined with autofluorescence of bone matrix (Fig. 5C-G). In the stump of amputated fins at 2 dpa, Zns5 immunoreactivity was increased at the tips of the bones, indicating the dedifferenatiation of osteoblasts (Fig. 5C). By contrast, at days 2 and 3 after cryoinjury, such a population of Zns5-positive osteoblasts was not observed (Fig. 5D-E). Indeed, the visualization of mature osteoblasts in transgenic reporter fish (osteocalcin:GFP) revealed destruction of bone-producing cells along the cryoinjured plane (Fig. S2). Thus, after cryoinjury, the regenerating margin needs to cope with the presence of the displaced matrix remnants of the destroyed bone segments.Fig. 5.


Zebrafish fin regeneration after cryoinjury-induced tissue damage.

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

Detachment of the destroyed fin tissue is associated with displacement and resorption of the dead bone fragments at the wound margin. (A,B) Imaging of bones in the same fin detected by autofluorescence of the mineralized matrix at 2 and 6 dpci. The margin of the remnant fins contains detached and displaced bone fragments between the rays that become resolved (arrows). N=5. (C-G) Confocal imaging of whole-mount fins immunostained with the osteoblast marker Zns5 (red), phagocyte marker L-plastin (green) and autofluorescent bone matrix (blue) at 2 dpa (C) and at different time points after cryoinjury (D-G). At 2 dpa (C), Zns5-labelled osteoblasts accumulate at the tip of the bone to initiate bone regeneration. L-plastin-expressing cells are present in the entire tissue. At 2 dpci (D) and 3 dpci (E), osteoblasts are scattered along the bones in irregular manner. At 5 dpci (F), Zns5-positive cells are enriched at the tips of the intact bones, below the margin of the stump that contains bone debris devoid of osteoblasts. At 7 dpci (G), Zns5 immunostaining is robustly enhanced along the remaining bones, indicating resumed regeneration. L-plastin-expressing cells are associated with the repairing and regenerating tissue. N=4. Scale bar in A=200 µm, in C=100 µm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4920184&req=5

BIO016865F5: Detachment of the destroyed fin tissue is associated with displacement and resorption of the dead bone fragments at the wound margin. (A,B) Imaging of bones in the same fin detected by autofluorescence of the mineralized matrix at 2 and 6 dpci. The margin of the remnant fins contains detached and displaced bone fragments between the rays that become resolved (arrows). N=5. (C-G) Confocal imaging of whole-mount fins immunostained with the osteoblast marker Zns5 (red), phagocyte marker L-plastin (green) and autofluorescent bone matrix (blue) at 2 dpa (C) and at different time points after cryoinjury (D-G). At 2 dpa (C), Zns5-labelled osteoblasts accumulate at the tip of the bone to initiate bone regeneration. L-plastin-expressing cells are present in the entire tissue. At 2 dpci (D) and 3 dpci (E), osteoblasts are scattered along the bones in irregular manner. At 5 dpci (F), Zns5-positive cells are enriched at the tips of the intact bones, below the margin of the stump that contains bone debris devoid of osteoblasts. At 7 dpci (G), Zns5 immunostaining is robustly enhanced along the remaining bones, indicating resumed regeneration. L-plastin-expressing cells are associated with the repairing and regenerating tissue. N=4. Scale bar in A=200 µm, in C=100 µm.
Mentions: To characterize the defects in cryoinjured fins, we analysed the morphology of the remaining bones after tissue sloughing. Live imaging of fins at 2 dpci revealed an accumulation of bone fragments at the margin of the stump (Fig. 5A). The tips of the bones were released from the rays and often displaced perpendicularly to the original position. Imaging of the same fins after the next 4 days (at 6 dpci) revealed that the remnants of mineralized matrix were undergoing degradation and resorption (Fig. 5B). To test whether the detached bone segments were associated with bone-producing cells, the osteoblasts, we performed immunofluorescence staining with Zns5 antibody and we labelled phagocytic cells with anti-L-plastin antibody of whole-mount fins combined with autofluorescence of bone matrix (Fig. 5C-G). In the stump of amputated fins at 2 dpa, Zns5 immunoreactivity was increased at the tips of the bones, indicating the dedifferenatiation of osteoblasts (Fig. 5C). By contrast, at days 2 and 3 after cryoinjury, such a population of Zns5-positive osteoblasts was not observed (Fig. 5D-E). Indeed, the visualization of mature osteoblasts in transgenic reporter fish (osteocalcin:GFP) revealed destruction of bone-producing cells along the cryoinjured plane (Fig. S2). Thus, after cryoinjury, the regenerating margin needs to cope with the presence of the displaced matrix remnants of the destroyed bone segments.Fig. 5.

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