Macrophages modulate adult zebrafish tail fin regeneration

Planarian flatworms regenerate their heads and tails from anterior or posterior wounds and this regenerative blastema polarity is controlled by Wnt/β-catenin signaling. It is well known that a regeneration blastema of appendages of vertebrates such as fish and amphibians grows distally. However, it remains unclear whether a regeneration blastema in vertebrate appendages can grow proximally. Here, we show that a regeneration blastema in zebrafish fins can grow proximally along the proximodistal axis by calcineurin inhibition. We used fin excavation in adult zebrafish to observe unidirectional regeneration from the anterior cut edge (ACE) to the posterior cut edge (PCE) of the cavity and this unidirectional regeneration polarity occurs as the PCE fails to build blastemas. Furthermore, we found that calcineurin activities in the ACE were greater than in the PCE. Calcineurin inhibition induced PCE blastemas, and calcineurin hyperactivation suppressed fin regeneration. Collectively, these findings identify calcineurin as a molecular switch to specify the PCE blastema of the proximodistal axis and regeneration polarity in zebrafish fin.

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Zebrafish kit mutation reveals primary and secondary regulation of melanocyte development during fin stripe regeneration

Development ◽

10.1242/dev.127.17.3715 ◽

2000 ◽

Vol 127 (17) ◽

pp. 3715-3724 ◽

Cited By ~ 10

Author(s):

J.F. Rawls ◽

S.L. Johnson

Keyword(s):

Receptor Tyrosine Kinase ◽

Mutational Analysis ◽

Adult Zebrafish ◽

Wild Type ◽

Kit Mutation ◽

Fin Regeneration ◽

Normal Melanocyte ◽

Kit Receptor ◽

Early Regeneration ◽

Secondary Mode

Fin regeneration in adult zebrafish is accompanied by re-establishment of the pigment stripes. To understand the mechanisms underlying fin stripe regeneration and regulation of normal melanocyte stripe morphology, we investigated the origins of melanocytes in the regenerating fin and their requirement for the kit receptor tyrosine kinase. Using pre-existing melanin as a lineage tracer, we show that most fin regeneration melanocytes develop from undifferentiated precursors, rather than from differentiated melanocytes. Mutational analysis reveals two distinct classes of regeneration melanocytes. First, an early regeneration class develops dependent on kit function. In the absence of kit function and kit-dependent melanocytes, a second class of melanocytes develops at later stages of regeneration. This late kit-independent class of regeneration melanocytes has little or no role in wild-type fin stripe development, thus revealing a secondary mode for regulation of fin stripes. Expression of melanocyte markers in regenerating kit mutant fins suggests that kit normally acts after mitf and before dct to promote development of the primary kit-dependent melanocytes. kit-dependent and kit-independent melanocytes are also present during fin stripe ontogeny in patterns similar to those observed during regeneration.

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Notch regulates blastema proliferation and prevents differentiation during adult zebrafish fin regeneration

Development ◽

10.1242/dev.087346 ◽

2013 ◽

Vol 140 (7) ◽

pp. 1402-1411 ◽

Cited By ~ 49

Author(s):

J. Munch ◽

A. Gonzalez-Rajal ◽

J. L. de la Pompa

Keyword(s):

Adult Zebrafish ◽

Fin Regeneration

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Impact of hypoxia induced VEGF and its signaling during caudal fin regeneration in Zebrafish

10.1101/105767 ◽

2017 ◽

Cited By ~ 1

Author(s):

sagayaraj.R Vivek ◽

R. Malathi

Keyword(s):

Wound Healing ◽

Blood Vessel ◽

Cobalt Chloride ◽

Adult Zebrafish ◽

Caudal Fin ◽

Fin Regeneration ◽

Blood Vessel Formation ◽

Vessel Formation ◽

Vegf Signaling ◽

Fin Length

ABSTRACT:Hypoxia is known to play important role during various cellular process, including regeneration. Regeneration is a complex process involving wound healing and tissue repair. We propose that hypoxia might mediate regeneration through angiogenesis involving angiogenic factors such as VEGF, VEGF-R2, NRP1a during the wound healing process. We have chosen Zebrafish model to study the role of hypoxia induced regeneration. Unlike mammals Zebrafish has the ability to regenerate. Hypoxic condition was mimicked using inorganic salt cobalt chloride to study caudal fin regeneration in adult Zebrafish. Intense blood vessel formation, with increased tail fin length experimented at various time points have been observed when adult zebrafish caudal fin partially amputated were exposed to 1% CoCl2. Regeneration is enhanced under hypoxia, with increased VEGF expression. To study the significance of VEGF signaling during wound healing and tissue regeneration, sunitinib well known inhibitor of VEGF receptor is used against CoCl2-induced caudal fin regeneration. Diminished fin length, lowering of blood vessel formation was documented using angioquant software, reduction in mRNA level of hypoxia inducible factors, VEGF and other pro-angiogenic genes such as VEGF, VEGF-R2, NRP1A, FGFR2, ANGPT1 were observed, while reduction in VEGF protein was demonstrated using western blot analysis. Genistein inhibitor of HIF-1α completely arrested regeneration, with suppression of VEGF highlighting the significance of hypoxia induced VEGF signaling during fin regeneration. Our results suggest that hypoxia through HIF-1α might lead to angiogenesis involving VEGF signaling during wound healing and this might throw light on therapeutic efficacy of cobalt chloride during regeneration.

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ScreenCube: A 3D Printed System For Rapid and Cost-Effective Chemical Screening in Adult Zebrafish

10.1101/163469 ◽

2017 ◽

Author(s):

Adrian T. Monstad-Rios ◽

Claire J. Watson ◽

Ronald Y. Kwon

Keyword(s):

Small Molecule ◽

Cost Effective ◽

Adult Zebrafish ◽

Fin Regeneration ◽

Drug Dosing ◽

Chemical Screening ◽

Models Of Development ◽

Dosing Regimens ◽

3D Printed ◽

Fish Survival

ABSTRACTPhenotype-based small molecule screens in zebrafish embryos and larvae have been successful in accelerating pathway and therapeutic discovery for diverse biological processes. Yet, the application of chemical screens to adult physiologies has been relatively limited due to additional demands on cost, space, and labor associated with screens in adult animals. Here, we present a 3D printed system and methods for intermittent drug dosing that enable rapid and cost-effective chemical administration in adult zebrafish. Using pre-filled screening plates, the system enables dosing of 96 fish in ˜3min, with a tenfold-reduction in drug quantity compared to that used in previous chemical screens in adult zebrafish. We characterize water quality kinetics during immersion in the system, and use these kinetics to rationally design intermittent dosing regimens that result in 100% fish survival. As a demonstration of system fidelity, we show the potential to identify two known chemical inhibitors of adult tail fin regeneration, cyclopamine and dorsomorphin. By developing methods for rapid and cost-effective chemical administration in adult zebrafish, this study expands the potential for small molecule discovery in post-embryonic models of development, disease, and regeneration.

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An early Shh-H2O2 feedback loop controls the progression of the regenerative program during adult zebrafish fin regeneration

10.1101/2021.08.19.456615 ◽

2021 ◽

Author(s):

Marion Thauvin ◽

Rodolphe Matias de Sousa ◽

Marine Alves ◽

Michel Volovitch ◽

Sophie Vriz ◽

...

Keyword(s):

Reactive Oxygen Species ◽

Sonic Hedgehog ◽

Feedback Loop ◽

Ros Production ◽

Oxygen Species ◽

Adult Zebrafish ◽

Regenerative Processes ◽

Fin Regeneration ◽

Spatiotemporal Regulation ◽

New Findings

AbstractReactive oxygen species (ROS), originally classified as toxic molecules, have attracted increasing interest given their actions in cell signaling. Among these molecules, Hydrogen peroxide (H2O2) is the major ROS produced by cells and acts as a second messenger to modify redox-sensitive proteins or lipids. After amputation, tight spatiotemporal regulation of ROS is required first for wound healing and later to initiate the regenerative program. However, the mechanisms carrying out this sustained ROS production and their integration with signaling pathways are still poorly understood. We focused on the early dialog between H2O2 and Sonic Hedgehog (Shh) during fin regeneration. We demonstrate that H2O2 controls Shh expression and that Shh in turn regulates the H2O2 level via a canonical pathway. Moreover, this tightly controlled feedback loop changes during the successive phases of the regenerative program. Dysregulation of the Hedgehog pathway has been implicated in several developmental syndromes, diabetes and cancer. These data support the existence of a very early feedback loop between Shh and H2O2 that might be more generally involved in various physiological or pathological processes. These new findings pave the way to improve regenerative processes, particularly in vertebrates.

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Macrophages modulate adult zebrafish tail fin regeneration

Development ◽

10.1242/dev.098459 ◽

2014 ◽

Vol 141 (13) ◽

pp. 2581-2591 ◽

Cited By ~ 146

Author(s):

T. A. Petrie ◽

N. S. Strand ◽

C. Tsung-Yang ◽

J. S. Rabinowitz ◽

R. T. Moon

Keyword(s):

Adult Zebrafish ◽

Fin Regeneration

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19-P025 Bone dynamics during adult zebrafish caudal fin regeneration

Mechanisms of Development ◽

10.1016/j.mod.2009.06.813 ◽

2009 ◽

Vol 126 ◽

pp. S298

Author(s):

Sara Sousa ◽

Nuno Afonso ◽

Mariana Fonseca ◽

António Jacinto

Keyword(s):

Adult Zebrafish ◽

Caudal Fin ◽

Fin Regeneration ◽

Bone Dynamics

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Faculty Opinions recommendation of Macrophages modulate adult zebrafish tail fin regeneration.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.718472419.793497049 ◽

2014 ◽

Author(s):

Guillermo Oliver ◽

Noelia Escobedo

Keyword(s):

Adult Zebrafish ◽

Fin Regeneration

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The roles of NADPH oxidases during adult zebrafish fin regeneration

10.1101/2021.07.13.452203 ◽

2021 ◽

Author(s):

Kunal Chopra ◽

Milda Folkmanaite ◽

Liam Stockdale ◽

Vishali Shathish ◽

Shoko Ishibashi ◽

...

Keyword(s):

Transgenic Line ◽

The Other ◽

Whole Body ◽

Ros Production ◽

Oxygen Species ◽

Adult Zebrafish ◽

Nadph Oxidases ◽

Fin Regeneration ◽

Organ Regeneration ◽

Mutant Lines

Sustained elevated levels of reactive oxygen species (ROS) have been shown to be essential for whole body, appendage and organ regeneration in various organisms, including planarians, Hydra, zebrafish, axolotl, Xenopus, geckos and mice. In the majority of cases these roles have been shown via the use of NADPH oxidase pharmacological inhibitors, which generally target all NAPDH oxidases (NOXes). To identify the specific NOX or NOXes essential for ROS production during adult fin regeneration in zebrafish, we generated nox mutants for duox, nox5 and cyba (a key subunit of NOXes 1-4). We also crossed these mutant lines to a transgenic line ubiquitously expressing HyPer, which permits the measurement of ROS levels in adult zebrafish fins. Using this approach, we found that homozygous duox mutants have significantly attenuated ROS levels following fin amputation, and this correlated with a significantly diminished rate of fin regeneration. While the other nox homozygous mutants (nox5 and cyba) showed less of an effect on ROS levels or adult fin regeneration, duox/cyba double mutants showed a more diminished rate of fin regeneration than duox mutants alone, suggesting that Nox1-4 do play a role during regeneration, but one that is secondary to that of Duox. This work also serendipitously found that ROS levels in amputated adult zebrafish fins oscillate during the day with a circadian rhythm.

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