Regeneration of Organs and Appendages in Zebrafish: A Window into Underlying Control Mechanisms

Abstract

The ability to regenerate organs and appendages is not universal among animals. Humans have a rather limited capacity to regenerate after an injury, while other vertebrates such as the zebrafish are capable of regenerating many anatomical structures. It is unknown why different vertebrates have such differences in regenerative capacity, so studying animal models that do regenerate will allow us to know what is needed to regenerate. Zebrafish research is not only able to tell us what mechanisms are involved in regeneration, but it also shows us that there are different regeneration strategies: some use stem cells while other create progenitors from differentiated tissue cells. This article details how zebrafish uses and regulates either differentiated tissue cells or stem cells to regenerate. We focus on four structures – fin appendage, brain, spinal cord and heart – and describe how current cell and molecular discoveries from these regenerating fish structures contribute to our understanding of general principles of regenerative biology.

Key Concepts

  • The zebrafish is a remarkable in vivo model to understand the stem/progenitor cell biology and molecular mechanisms involved in appendage and organ regeneration.
  • Studying how the zebrafish regenerates its fins, its nervous system and its heart will provide information on how endogenous cell populations (whether fully differentiated or stem cells) are activated and concertedly controlled to regenerate compound structures.
  • Different zebrafish tissues use different strategies to regenerate: some convert differentiated tissue cells to progenitor cells while other draw from stem cell pools.
  • Zebrafish appendages and hearts regenerate primarily from the residual differentiated tissues.
  • Zebrafish nervous tissue regenerates from resident stem cell populations.

Keywords: zebrafish; regeneration; fin; brain; spinal cord; heart; stem cells; progenitor cells

Figure 1. Regeneration of the caudal fin in adult and larval zebrafish. Regeneration of the adult fin. (a) The adult caudal fin is a bi‐lobed appendage whose main structural elements are the segmented (J, joints) ray bones (lepidotrichia) (l). These rays bifurcate (B) at set positions. (b) Amputation induces epidermal migration over the injury. After wound closure, a thickened wound epidermis (We, yellow) forms. (c) Subsequently, cells underneath begin to proliferate at the distal tips of the fin rays to form blastemas of the rays (Blray, dark red) and interrays (Blinterray, red). (d) As the blastema cells continue to proliferate, new tissues are generated at the proximal end of the blastema. (e) Outgrowth continues until the fin has reached its original length. Regeneration of the larval finfold. (f) The tail of the larva contains several tissues: the notochord (nc) and neural tube (nt) are surrounded by somatic skeletal muscle (sm). These tissues are enclosed by an overlying epithelial layer that extends ventrally and dorsally along the body and posteriorly at the end of the tail to form an epithelial finfold (ff). The skeletal structure is a scaffold of actinotrichia (A). Pigment cells (P) run dorsally and ventrally along the body. (g) Amputation of the finfold results in wound closure. (h) Regeneration of the fold involves cell proliferation at the distal edge, and this results in regenerative outgrowth of the fold. (i) As this process continues, pigment cells migrate distally. This outgrowth process continues until the regenerate resembles a fin fold. Cross section through regenerating adult fin ray. (j) Cross section through a regenerating fin ray shows the accumulated cells of the blastema (Bl) surrounded by the pro‐regenerative wound epidermis (We). Cells at the proximal interface between the blastema and the tissues of the stump contribute as newly differentiated tissues: osteoblasts (O), fibroblasts (f), segmented lepidotrichia (l), nerve (n), vasculature (v). (k) Diagrams of the expression patterns of the ligand and receptors in different signal transduction pathways. (l) Positional expression of genes along the anterio–posterior (A–P) axis of the pectoral fin.
Figure 2. Neuron regeneration in the zebrafish brain and spinal cord. (a) The adult zebrafish brain (anterior pointing left) has precisely arranged regions that contain cells that continually proliferate and are considered to be stem cell niches (red). Around these regions, neurogenic zones (dark blue) form where cells from the stem cell niches migrate and differentiate into new neurons. Vertical bars present the sections through the zebrafish brain shown in panels (b), (c) and (d). (b) One model for the continued growth of the adult brains is that stem cell niches surrounding the ventricle produce neuronal progenitors, and that these progenitors differentiate to form new neurons (new growth) along the periphery of the brain. The continued accumulation of the differentiated progeny of the progenitors along the periphery of the brain results in constant growth. (c) PCNA staining (blue) of cross‐sections through the cerebellum of an adult transgenic : zebrafish brain, illustrating the structure of the adult neural stem cell niche in the cerebellum. The gene is a marker for neuronal progenitors. Insert provides greater resolution of doubly labeled : ‐PCNA‐positive cells. The plane of the cross‐section through the brain is depicted by the white diagram in lower left corner. (d) Staining of cross‐section through the ventral telencephalon (V) with PCNA (red fluorescence) identifies a zone of proliferation along the ventral and lateral ventricle, and the retention of BrdU label (green fluorescence) after pulse‐chase experiments indicates that after proliferation, these cells migrate into the ventricle to become neurons. Cells double‐positive for BrdU and PCNA (yellow fluorescence, white arrow) are located only along the ventricular space, indicating that the BrdU‐labelled cells throughout the section originate from proliferation zone. The plane of the cross‐section through the brain is depicted by the white diagram in upper right corner. (e) Stab injury to the telencephalon induces an inflammatory response around the wound (brown circles), which leads to cell proliferation (neurogenesis: green) around the injury site and in the sub‐ventricular zones on the periphery of the injured lobe. (f) Subsequent migration of neuronal axons from the surrounding uninjured telencephalon tissue. (g) Regeneration in the telencephalon lobe restores the tissues to their original homeostasis. (h) The zebrafish spinal cord (Sc) runs from the brain (Br) to the base of the caudal fin. As for all vertebrates, transection of the spinal cord results in paralysis caudal to the transection site. (i) Histological cross‐sections through the un‐lesioned spinal cord show different nerve types and a narrow ventricle; BrdU incorporation studies indicate that cells do not cycle. However, after injury to the spinal cord, cells along the ventricle do incorporate BrdU (green). Cell tracking of BrdU‐labelled cells indicates that these cells migrate from the ventricular zone. Furthermore, immunohistochemistry with markers for neuronal differentiation (ChAT) and synapse formation (SV) indicate that these cells become functional neurons. (j) Over time, the new nerves extend over the transection site, and 27–47% of these are neuronal extensions from the brain, as demonstrated by the anterograde transport of vital dye (indicated green) past the transected site of spinal cord when injected into cell bodies in the brainstem. (k) Neuronal connections in the spinal cord also transport vital dye (green) retrograde into neurons in the brain; however, this occurs significantly less frequently. Only 0.3–1.2% of the neurons regenerate ascending axons into the brain stem. (l) Transection through the spinal cord results in the production and release of dopamine (red dots) from dopaminergic neurons (red). The dopaminergic receptors (D4) are expressed by the ependymal stem cells bordering the sub‐ventricular zone running along the centre of the spinal cord. These ependymal cells proliferate to produce progenitor cells that generate motor neurons. 0 and 14 dpt are days post transection. The scale bars in panels (c), (d) equal 50 µm. (c) Reproduced with Permission from Kaslin et al. (2009) © Society of Neuroscience. (d) Reproduced from Grandel et al. (2006) © Elsevier.
Figure 3. Heart regeneration. (a) The zebrafish heart contains two chambers: one atrium (A) and one ventricle (V). The bulbous arteriosus (B) is the outflow tract of the heart. The ventricle consists of three tissue layers: the epicardium (Epi, red), myocardium (Myo, pink) and the endocardium (En, dark blue). The epicardium lines the periphery of the myocardium. The myocardium consists of different muscle layers: the primordial muscle layer (PrM), the trabeculated muscle layer (Tr, depicted perpendicular to the ventricular wall) and the cortical muscle layer (CrM, depicted parallel to the ventricular wall). The different coloured myocardial cells represent muscle cell populations derived from different progenitor cell clones during cardiac development. The tissue lining the interior of the chamber is the endocardium. The red arrows indicate the direction of blood flow. (b) Zebrafish can lose up to 20% of its ventricular muscle after amputation injury (dashed line). (c) Immediately after amputation, a durable clot forms to prevent further blood loss from the opened chamber of the ventricle. (d) New ventricular tissue regenerates from the residual ventricle. The epicardium expresses , , , , and and migrates over the scar, while the myocardium expresses the ligands , , , and . Cortical muscle cells proliferate and begin to replace the scar. The migrating cortical muscle expresses (blue circular nuclei). The endocardium also begins to migrate into the wound and expresses . (e) This regeneration process gradually replaces the scar from all sides of the injury to form a thickened ventricular muscle wall where the amputation took place. (f) Over time, remodelling returns the ventricular wall to its original dimensions. The new ventricular wall consists of a thickened wall of myocardium from the cortical muscle and a single layer of cells from the primordial layer. Scissors in panels (c–f) indicate the plane of amputation.
close

References

Adolf B, Chapouton P, Lam CS, et al. (2006) Conserved and acquired features of adult neurogenesis in the zebrafish telencephalon. Developmental Biology 295: 278–293.

Aguirre A, Montserrat N, Zacchigna S, et al. (2014) In vivo activation of a conserved microRNA program induces mammalian heart regeneration. Cell Stem Cell 15: 589–604.

Akimenko M‐A, Mari‐Beffa M, Becerra J, et al. (2003) Old questions, new tools, and some answers to the mystery of fin regeneration. Developmental Dynamics 226: 190–201.

Alunni A, Hermel J‐M, Heuze A, et al. (2010) Evidence for neural stem cells in the Medaka optic tectum proliferation zones. Developmental Neurobiology 70: 693–713.

Alunni A, Krescmarik M, Bosco A, et al. (2013) Notch3 signaling gates cell cycle entry and limits neural stem cell amplification in the adult pallium. Development 140: 3335–3347.

Anchelin M, Murcia L, Alcaraz‐Perez F, et al. (2011) Behaviour of telomere and telomerase during aging and regeneration in zebrafish. PLoS One 6: e16955.

Antos CL and Tanaka EM (2010) Vertebrates that regenerate as models for guiding stem cells (The Cell Biology of Stem Cells. E. Meshorer and K Plath, Landes Bioscience). Advances in Experimental Medicine and Biology 695: 184–214.

Azevedo AS, Grotek B, Jacinto A, et al. (2011) The regenerative capacity of the zebrafish caudal fin is not affected by repeated amputations. PLoS One 6: e22820.

Balak KJ, Corwin JT and Jones JE (1990) regenerated hair cells can originate from supporting cell progeny: evidence from phototoxicity and laser ablation experiments in the lateral line system. Journal of Neuroscience 10: 2502–2512.

Barnabe‐Heider F, Göritz C, Sabelström H, et al. (2010) Origin of new glial cells in intact and injured adult spinal cord. Cell Stem Cell 7: 470–482.

Bayliss PE, Bellavance KL, Whitehead GG, et al. (2006) Chemical modulation of receptor signaling inhibits regenerative angiogenesis in adult zebrafish. Nature Chemical Biology 2: 265–273.

Becker T, Bernhardt RR, Reinhard E, et al. (1998) Readiness of zebrafish brain neurons to regenerate a spinal axon correlates with differential expression of specific cell recognition molecules. Journal of Neuroscience 18: 5789–5803.

Becker CB and Becker T (2008) Adult zebrafish as a model for successful central nervous system regeneration. Restorative Neurology and Neuroscience 26: 72–80.

Berg DA, Kirkham M, Wang H, et al. (2011) Dopmaine controls neurogenesis in the adult salamander midbrain in homeostasis and during regeneration of dopamine neurons. Cell Stem Cell 8: 426–433.

Bernardos RL, Barthel LK, Meyers JR, et al. (2007) Late‐stage neuronal progenitors in the retina are radial mueller glia that function as retinal stem cells. Journal of Neuroscience 27: 7028–7040.

Bhatt DH, Otto SJ, Depoister B, et al. (2004) Cyclic AMP‐induced repair of zebrafish spinal circuits. Science 305: 254–258.

Blum N and Begemann G (2012) Retinoic acid signaling controls the formation, proliferation and survival of the blastema during adult zebrafish fin regeneration. Development 139: 107–116.

Boccalini G, Sassoli C, Formigli L, et al. (2015) Relaxin protects cardiac muscle cells from hypoxia/reoxygenation injury: involvement of the Notch‐1 pathway. FASEB Journal 29: 239–249.

Borday V, Thaeron C, Avaron F, et al. (2001) evx1 transcription in bony fin rays segment boundaries leads to a reiterated pattern during zebrafish fin development and regeneration. Developmental Biology 220: 91–98.

Briata P, Van DeWerken R, Airoldi I, et al. (1995) Transcriptional repression by the human homeobox protein EVX1 in transfected mammalian cells. Journal of Biological Chemistry 270: 27695–27701.

Briona LK and Dorsky RL (2014) Radial Glial progenitors repair the zebrafish spinal cord following transection. Experimental Neurology 256: 81–92.

Chablais F and Jazwinska A (2010) IGF signaling between blastema and wound epidermis is required for fin regeneration. Development 137: 871–879.

Chablais F, Veit J, Rainer G, et al. (2011) The zebrafish hear regenerates after cyroinjury‐induced myocardial infarction. BMC Developmental Biology 11: 21.

Chablais F and Jazwinska A (2012) The regenerative capacity of the zebrafish heart is dependent on TGFβ signaling. Development 139: 1921–1930.

Chapouton P, Skupien P, Hesl B, et al. (2010) Notch activity levels control the balance between quiescence and recruitment of adult neural stem cells. Journal of Neuroscience 30: 7961–7974.

Chen C‐H, Durand E, Wang J, et al. (2013) Zebrafish transgenic lines for in vivo bioluminescence imaging of stem cells and regeneration in adult zebrafish. Development 140: 4988–4997.

Chera S, Ghila L, Dobretz K, et al. (2009) Apoptotic cells provide an unexpected source of Wnt3 signaling to drive Hydra head regeneration. Developmental Cell 17: 279–289.

Choi W‐Y, Gemberling M, Wang J, et al. (2013) In vivo monitoring of cardiomyocyte proliferation to identify chemical modifiers of heart regeneration. Development 140: 660–666.

Clubb FJ Jr and Bishop SP (1984) Formation of binucleated myocardial cells in the neonatal rat. An index for growth hypertrophy. Laboratory Investigation 50: 571–577.

Diotel N, Vaillant C, Gueguen M‐M, et al. (2010) Cxcr4 and cxcl12 expression in radial glial cells of the brain of adult zebrafish. Journal of Comparative Neurology 518: 4855–4878.

Dufourcq P and Vriz S (2006) The chemokine SDF‐1 regulates blastema formation during zebrafish fin regeneration. Development Genes and Evolution 216: 635–639.

Fang Y, Gupta V, Karra R, et al. (2013) Translational profiling of cardiomyocytes identifies an early Jak1/Stat3 injury response required for zebrafish heart regeneration. Proceedings of the National Academy of Sciences of the United States of America 110: 13416–13421.

Ganz J, Kaslin J, Hochmann S, et al. (2010) Heterogeneity and Fgf dependence of adult neural progenitors in the zebrafish telencephalon. Glia 58: 1345–1363.

Gauron C, Rampon C, Bouzaffour M, et al. (2013) Sustained production of ROS triggers compensatory proliferation and is required for regeneration to proceed. Scientific Reports 3: 2084.

Geraudie J and Singer M (1985) Necessity of an adequate nerve supply for regeneration of the amputated pectoral fin in the teleost Fundulus. Journal of Experimental Zoology 234: 367–374.

Gerhard GS, Kauffman EJ, Wang X, et al. (2002) Life spans and senescent phenotypes in two strains of zebrafish (Danio rerio). Experimental Gerontology 37: 1055–1068.

Goldshmit Y, Sztal TE, Jusuf PR, et al. (2012) Fgf‐dependent glial cell bridges facilitate spinal cord regeneration in zebrafish. Journal of Neuroscience 32: 7477–7492.

Gonzalez‐Rosa JM, Peralta M and Mercader N (2012) Pan‐epicardial lineage tracing reveals that epicardium derived cells give rise to myofibroblasts and perivascular cells during zebrafish heart regeneration. Developmental Biology 370: 173–186.

Gonzalez‐Rosa JM, Guzman‐Martinez G, Marques IJ, et al. (2014) Use of echocardiography reveals reestablishment of entricular pumping efficiency and partial ventricular wall motion recovery upon ventricular cryoinjury in the zebrafish. PLoS One 9 (12): e115604.

Goss RJ and Stagg MW (1957) The regeneration of fins and fin rays in Fundulus heteroclitus. Journal of Experimental Zoology 136: 487–507.

Grandel H, Kaslin J, Ganz J, et al. (2006) Neural stem cells and neurogenesis in the adult zebrafish brain: origin, proliferation dynamics, migration and cell fate. Developmental Biology 295: 263–277.

Grotek B, Wehner D and Weidinger G (2013) Notch signaling coordinates cellular proliferation with differentiation during zebrafish fin regeneration. Development 140: 1412–1423.

Guo Y, Ma L, Cristofanilli M, et al. (2010) Transcription factor sox11b is involved in spinal cord regeneration in adult zebrafish. Journal of Neuroscience 172: 329–341.

Gupta V and Poss KD (2012) Clonally dominant cardiomyocytes direct heart morphogenesis. Nature 484: 479–484.

Gupta V, Gemberling M, Karra R, et al. (2013) An injury‐responsive Gata4 program shapes the zebrafish cardiac ventricle. Current Biology 23: 1221–1227.

Hanna GFB, Nawar NN and Sharma SC (1998) Regeneration fo ascending spinal axons in goldfish. Brain Research 791: 235–245.

Hein S, Lehmann LH, Kossack M, et al. (2015) Advanced echocardiography in adult zebrafish reveals delayed recovery of heart function after myocardial cryoinjury. PLoS One. DOI: 10.1371/journal.pone.0122665.

Hirose K, Shimoda N and Kikuchi Y (2013) Transient reduction of 5‐methylcytosine and 5‐hydroxymethylcytosine is associated with active DNA demethylation during regenration of zebrafish fin. Epigenetics 8: 899–906.

Höglinger GU, Rizk P, Muriel MP, et al. (2004) Dopamine depletion impairs precursor cell proliferation in Parkinson disease. Nature Neuroscience 7: 726–735.

Hoptak‐Solga AD, Klein KA, DeRosa AM, et al. (2007) Zebrafish short fin mutations in connexin43 lead to aberrant gap junctional intercellular communication. FEBS Letters 581 (17): 3297–3302.

Hu G, Lee H, Price SM, et al. (2001a) Msx homeobox genes inhibit differentiation through upregulation of cyclin D1. Development 128: 2373–2384.

Hu N, Yost HJ and Clark EB (2001b) Cardiac mophology and blood pressure in the adult zebrafish. Anatomical Record 264: 1–12.

Huang Y, Harrison MR, Osorio A, et al. (2013) Igf signaling is required for cardiomyocyte proliferation during zebrafish heart development and regeneration. PLoS One 8 (6): e67266.

Huang C‐C, Su T‐H and Shih C‐C (2015) High‐resolution tissue doppler imaging of the zebrafish heart during its regeneration. Zebrafish 12: 48–57.

Ito Y, Tanaka H, Okamoto H, et al. (2010) Characterization of neural stem cells and their progeny in the adult zebrafish optic tectum. Developmental Biology 342: 26–38.

Jazwinska A, Badakov R and Keating MT (2007) Activin‐βA signaling is required for zebrafish fin regeneration. Current Biology 17: 1390–1395.

Jopling C, Sleep E, Raya M, et al. (2010) Zebrafish heart regeneration occurs by cardiomyocyte dedifferentiation and proliferation. Nature 464: 606–610.

Kametani Y, Chi NC, Stainier DYR, et al. (2015) Notch signaling regulates venous arterialization during zebrafish fin regeneration. Genes to Cells 20 (5): 427–438. DOI: 10.1111/gtc.12234.

Kanaoka Y and Boyce JA (2004) Cysteinyl leukotrienes and their receptors: cellular distribution and function in immune and inflammatory responses. Journal of Immunology 173: 1503–1510.

Kang J, Nachtrab G and Poss KD (2013) Local Dkk1 crosstalk from breeding ornaments impedes regeneration of injured male zebrafish fins. Developmental Cell 27: 19–31.

Kaslin J, Ganz J and Brand M (2008) Proliferation, neurogenesis and regeneration in the non‐mammalian vertebrate brain. Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences 363: 101–122.

Kaslin J, Ganz J, Geffarth M, et al. (2009) Stem cells in the adult zebrafish cerebellum: initiation and maintenance of a novel stem cell niche. Journal of Neuroscience 29: 6142–6153.

Kawakami A, Fukazawa T and Takeda H (2004) Early fin primordia of zebrafish larvae regenerate by a similar growth control mechanism with adult regeneration. Developmental Dynamics 231: 693–699.

Kikuchi K, Holdaway JE, Werdich AA, et al. (2010) Primary contribution to zebrafish heart regeneration by gata4+ cardiomyocytes. Nature, in press.

Kikuchi K, Gupta V, Wang J, et al. (2011a) tcf21+ epicardial cells adopt non‐myocardial fates during zebrafish heart development and regeneration. Development 138: 2895–2902.

Kikuchi K, Holdway JE, Major RJ, et al. (2011b) Retinoic acid production by endocardium function and epicardium is an injury response essential for zebrafish heart regeneration. Developmental Cell 20: 397–404.

Kirsche W (1961) Experimentelle Untersuchungen zur Frage der Regeneration und Funktion des Tectum Opticum von Carassius carassius. L. Z. Mikrosk. Anat. Forsch. 67: 140–182.

Kirsche W (1983) The significance of matrix zones for brain regeneration and brain transplantation with special consideration of lower vertebrates. In: Wallace RB and Das GD (eds) Neural Tissue Transplantation Research, pp. 65–104. New York: Springer‐Verlag.

Kizil C, Otto GW, Geisler R, et al. (2009) Simplet controls cell proliferation and gene transcription during zebrafish caudal fin regeneration. Developmental Biology 325: 329–340.

Kizil C, Kaslin J, Kroehne V, et al. (2011) Adult neurogenesis and brain regeneration in zebrafish. Developmental Neurobiology 72: 429–461.

Kizil C, Dudczig S, Kyritsis N, et al. (2012a) The chemokine receptor cxcr5 regulates the regenerative neurogenesis response in the adult zebrafish brain. Neural Development 7: 27.

Kizil C, Kyritsis N, Dudczig S, et al. (2012b) Regenerative neurogenesis from neural progenitor cells requires injury‐induced expression of Gata3. Developmental Cell 23: 1230–1237.

Knopf F, Hammond C, Chekuru A, et al. (2011) Bone regeneration via dedifferentiation of osteoblasts in the zebrafish fin. Developmental Cell 20: 725–732.

Koppanyi T and Weiss P (1922) Funktionelle regeneration des Rückenmarkes bei Anamniern. Anz. Akad. Wiss. Wien Math. Naturw. Kl. 59: 206.

Kragl M, Knapp D, Nacu E, et al. (2009) Cells keep a memory fo their tissue origin during axolotl limb regeneration. Nature 460: 60–65.

Kroehne V, Freudenreich D, Hans S, et al. (2011) Regeneration of the adult zebrafish brain from neurogenic radial glia‐type progenitors. Development 138: 4831–4841.

Kujawski S, Lin W, Kitte F, et al. (2014) Calcineurin regulates coordinated outgrowth of zebrafish regenerating fins. Developmental Cell 28: 1–15.

Kumar A, Velloso CP, Imokawa Y, et al. (2004) The regenerative plasticity of isolated urodele myofibers and its dependence on Msx1. PLoS Biology 2: 1168–1176.

Kyritsis N, Kizil C, Zocher S, et al. (2012) Acute inflammation initiates the regenerative response in the adult zebrafish brain. Science 338: 1353–1356.

Kyritsis N, Kizil C and Brand M (2014) Neuroinflammation and central nervous system regeneration in vertebrates. Trends in Cell Biology 24: 128–135.

Laflamme MA and Murry CE (2011) Heart regeneration. Nature 473: 326–335.

Laforest L, Brown CW, Poleo G, et al. (1998) Involvement of the sonic hedgehog, patched1 and bmp2 genes in patterning of the zebrafish dermal fin rays. Development 125: 4175–4184.

Lavine KJ, Yu K, White AC, et al. (2005) Endocardial and epicardial derived FGF signals regulate myocardial proliferation and differentiation in vivo. Developmental Cell 8: 85–95.

Lee H, Habas R and Abate‐Shen C (2004) Msx1 cooperates with histone H1b for inhibition of transcription and myogenesis. Science 304: 1675–1678.

Lee Y, Grill S, Sanchez A, et al. (2005) Fgf signaling instructs position‐dependent growth rate during zebrafish fin regeneration. Development 132: 5173–5183.

Lee Y, Hami D, Val SD, et al. (2009) Maintenance of blastemal proliferation by functionally diverse epidermis in regenerating zebrafish fins. Developmental Biology 331: 270–280.

Lepilina A, Coon AN, Kikuchi K, et al. (2006) A dynamic epicardial injury response supports progenitor cell activity during zebrafish heart regeneration. Cell 127: 607–619.

Lien C‐L, Schebesta M, Makino S, et al. (2006) Gene expression analysis of zebrafish heart regeneration. PLoS Biology 4: 1386–1396.

Lin J‐F, Pan H‐C, Ma L‐P, et al. (2012) The cell neural adhesion molecule contractin‐2 (TAG‐1) is beneficial for functional recovery after spinal cor injury in adult zebrafish. PLoS One 7: e52376.

Lund TC, Glass TJ, Tolar J, et al. (2009) Expression of telomerase and telomere length are unaffected by either age or limb regeneration in Danio rerio. PLoS One 4: e7688.

März M, Chapouton P, Diotel N, et al. (2010) Heterogeneity in progenitor cell subtypes in the ventricular zone of the zebrafish adult telencephalon. Glia 58: 870–888.

Mason I (2007) Initiation to end point: the multiple roles of fibroblast growth factors in neural development. Nature Review Neuroscience 8: 583–596.

Meletis K, Barnabe‐Heider F, Carlen M, et al. (2014) Spinal cord injury reveals multilineage differentiation of ependymal cells. PLoS Biology 6: e182.

Mercer SE, Odelberg SJ and Simon H‐G (2013) A dynamic spatiotemporal extracellular matrix facilities epicardial‐mediated vertebrate heart regeneration. Developmental Biology 382: 457–469.

Missale C, Nash SR, Robinson SW, et al. (1998) Dopamine receptors: from structure to function. Physiological Reviews 78: 189–225.

Monteiro J, Aires R, Becker JD, et al. (2014) V‐ATPase proton pumping activity is required for adult zebrafish appendage regeneration. PLoS One 9: e92594.

Münch J, Gonzalez‐Rajal A and Pompa JLdl (2013) Notch regulates blastema proliferation and prevents differentiation during adult zebrafish fin regeneration. Development 140: 1402–1411.

Murciano C, Ruiz J, Maseda D, et al. (2001) Ray and inter‐ray blastemas interact to control bifurcations of Danio rerio fin rays. International Journal of Developmental Biology 45: S129–S130.

Murciano C, Fernandez TD, Duran I, et al. (2002) Ray‐interray interactions during fin regeneration of Danio rerio. Developmental Biology 252: 214–224.

Murciano C, Perez‐Claros J, Smith A, et al. (2007) Position dependence of hemiray morphogenesis during tail fin regeneration in Danio rerio. Developmental Biology 312: 272–283.

Nachtrab G, Kikuchi K, Tornini VA, et al. (2013) Transcriptional components of anteroposterior positional information during zebrafish regeneration. Development 140: 3754–3764.

Narita T, Sasaoka S, Udagawa K, et al. (2005) Wnt10a is involved in AER formation during chick limb development. Developmental Dynamics 233: 282–287.

Neithammer P, Grabher C, Look AT, et al. (2009) A tissue‐scale gradient of hydrogen peroxide mediates rapid wound detection in zebrafish. Nature 459: 996–1000.

Ogai K, Nakatani K, Hisano S, et al. (2014) Function of Sox2 in ependymal cells of lesioned spinal cords in adult zebrafish. Neuroscience Research 88: 84–87.

de Oliviera‐Carlos V, Ganz J, Hans S, et al. (2013) Notch receptor expression in neurogenic regions of the adult zebrafish brain. PLoS One 8 (9): e73384.

Panman L and Zeller R (2003) Patterning the limb before and after SHH signalling. Journal of Anatomy 202: 3–12.

Pellegrini E, Mouriec K, Anglade I, et al. (2007) Identification of aromastase‐positive radial glial cells as progenitor cells in the ventricular layer of the forebrain in zebrafish. Journal of Comparative Neurology 501: 150–167.

Perathoner S, Daane JM, Henrion U, et al. (2014) Bioelectric signaling regulates size in zebrafish fins. PLoS Genetics 10: e1004080.

Petrie TA, Strand NS, Tsung‐Yang C, et al. (2014) Macrophages modulate adult zebrafish tail fin regeneration. Development 141: 581–2591.

Pfefferli C, Müller F, Jazwinsky A, et al. (2014) Specific NuRD components are required for fin regeneration in zebrafish. BMC Biology 12: 1–17.

Poleo G, Brown CW, Laforest L, et al. (2001) Cell proliferation and movement during early fin regeneration in zebrafish. Developmental Biology 221: 380–390.

Poss KD, Shen J, Nechiporuk A, et al. (2000) Roles for Fgf signaling during zebrafish fin regeneration. Developmental Biology 222: 347–358.

Poss KD, Nechiporuk A, Hillam AM, et al. (2002a) Mps1 defines a proximal blastema proliferative compartment essential for zebrafish fin regeneration. Development 129: 5141–5149.

Poss KD, Wilson LG and Keating MT (2002b) Heart regeneration in zebrafish. Science 298: 2188–2190.

Quint E, Smith A, Avaron F, et al. (2002) Bone patterning is altered in the regenerating zebrafish caudal fin after ectopic expression of sonic hedgehog and bmp2b or exposure to cyclopamine. Proceedings of the National Academy of Sciences of the United States of America 99: 8713–8718.

Raible F and Brand M (2004) Divide et Impera – the midbrain‐hindbrain boundary and its organizer. Trends in Neurosciences 27: 727–734.

Raya A, Koth CM, Buescher D, et al. (2003) Activation of notch signaling pathway precedes heart regeneration in zebrafish. Proceedings of the National Academy of Sciences of the United States of America 100: 11889–11895.

Reimer MM, Sörensen I, Kuscha V, et al. (2008) Motor neuron regeneration in adult zebrafish. Journal of Neuroscience 28: 8510–8516.

Reimer MM, Norris A, Ohnmacht J, et al. (2013) Dopamine from the brain promotes spinal motor neuron generation during development and adult regeneration. Developmental Cell 25 (5): 478–491.

Rolland‐Lagan A‐G, Paquette M, Tweedle V, et al. (2012) Morphogen‐based simulation model of ray growth and joint patterning during fin development and regeneration. Development 139: 1188–1197.

Sallin P, de Preux Charles A‐S, Duruz V, et al. (2015) A dual epimorphic and compensatory mode of heart regeneration in zebrafish. Developmental Biology 399 (1): 27–40.

Saneyoshi T, Kume S, Amasaki Y, et al. (2002) The Wnt/calcium pathway activates NF‐AT and promotes ventral cell fate in Xenopus embryos. Nature 417: 295–299.

Santos‐Ruiz L, Santamaria JA, Ruiz‐Sanchez J, et al. (2002) Cell proliferation during blastema formation in the regenerating teleost fin. Developmental Dynamics 223: 262–272.

Schindler YL, Garske KM, Wang J, et al. (2014) Hand2 elevates cardiomyocyte production during zebrafish heart development and regeneration. Development 141: 3112–3122.

Schulte CJ, Allen C, England SJ, et al. (2011) Evx1 is required for joint formation in zebrafish fin dermoskeleton. Developmental Dynamics 240: 1240–1248.

Shao J, Chen D, Ye Q, et al. (2011) Tissue regeneration after injury in adult zebrafish: the regenerative potential of the caudal fin. Developmental Dynamics 240: 1271–1277.

Simoes MG, Bensimon‐Brito A, Fonseca M, et al. (2014) Denervation impairs regeneratino of amputated zebrafish fins. BMC Developmental Biology. DOI: 10.1186/s12861-014-0049-2.

Singer M (1952) The influence of the nerve in regeneration of the amphibian extremity. Quarterly Review of Biology 27: 169–200.

Singh SP, Holdway JE and Poss KD (2012) Regeneration of amputated zebrafish fin rays from de novo osteoblasts. Developmental Cell 22: 879–886.

Smith A, Avaron F, Guay D, et al. (2006) Inhibition of BMP signaling during zebrafish fin regeneration disrupts fin growth and scleroblast differentiation and function. Developmental Biology 299: 438–454.

Sousa S, Afonso N, Bensimon‐Brito A, et al. (2011) Differentiated skeletal cells contribute to blastema formation during zebrafish fin regeneration. Development 138: 3897–3905.

Sousa S, Valerio F, Jacinto A, et al. (2012) A new zebrafish bone crush injury model. Biology Open 1 (9): 915–921.

Stewart S and Stankunas K (2012) Limited dedifferentiation provides replacement tissue during zebrafish fin regeneration. Developmental Biology 365: 339–349.

Stewart S, Gomez AW, Armstrong BE, et al. (2014) Sequential and opposing activities of Wnt and BMP coordinated zebrafish bone regeneration. Cell Reports 6: 482–498.

Stoick‐Cooper CL, Weidinger G, Riehle KJ, et al. (2007) Distinct Wnt signaling pathways have opposing roles in appendage regeneration. Development 134: 479–489.

Sugiura T, Tazaki A, Ueno N, et al. (2009) Xenopus Wnt‐5a induces an ectopic larval tail at injured site, suggesting a crucial role for noncanonical wnt signal in tail regeneration. Mechanisms of Development 126: 56–67.

Takayama K, Shimoda N, Takanaga S, et al. (2014) Expression patterns of dnmt3aa, dnmt3ab and dnmt4 during development and fin regeneration in zebrafish. Gene Expression Patterns 14: 105–110.

Thummel R, Bai S, Michael P, et al. (2006) Inhibition of zebrafish fin regeneration using in vivo electroporation of morpholinos against fgfr1 and msxb. Developmental Dynamics 235: 335–346.

Thummel R, Ju M, Michael P, et al. (2007) Both Hoxc13 orthologs are functinally important for zebrafish tail fin regeneration. Development Genes and Evolution 217: 413–420.

Ton QV and Iovine MK (2013) Identification of an evx‐1‐dependent joint formatino pathway during fin regeneration. PLoS One 8: e81240.

Topp S, Stigloher C, Komisarczuk AZ, et al. (2008) Fgf signaling in the zebrafish adult brain: association of Fgf activity with ventricular zones but not cell proliferation. Journal of Comparative Neurology 510: 422–439.

Tu S and Johnson SL (2011) Fate restriction in the growing and regenerating zebrafish fin. Developmental Cell 20: 725–732.

Tuge H and Hanazawa S (1935) Physiology of the spinal fish, with special reference to the postural mechanism. Science Reports of the Tohoku Imperial University. Series 4, Biology 10: 589–606.

Viales RR, Diotel N, Ferg M, et al. (2014) The helix‐loop‐helix protein Id1 controls stem cell proliferation during regenerative neurogenesis in the adult zebrafish telencephlon. Stem Cells 33 (3): 892–903. DOI: 10.1002/stem.1883.

Wang J, Panakova D, Kikuchi K, et al. (2011) The regenerative capacity of zebrafish reverses cardiac failure caused by genetic cardiomyocyte depletion. Development 138: 3421–3430.

Wang J, Karra R, Dickson AL, et al. (2013) Fibronectin is deposited by injury‐activated epicardial cells and is necessary for zebrafish heart regeneration. Developmental Biology 382: 427–435.

Wehner D, Cizelsky W, Vasudevaro MD, et al. (2014) Wnt/b‐catenin signaling defines organizing centers that orchestrate growth and differentiation of the regenerating zebrafish caudal fin. Cell Reports 6: 467–481.

Whitehead GG, Makino S, Lien C‐L, et al. (2005) fgf20 is essential for initiating zebrafish fin regeneration. Science 310: 1957–1960.

Wills AA, Holdway JE, Major RJ, et al. (2007) Regulated addition of new myocardial and epicardial cells fosters homeostatic cardiac growth and maintenance in adult zebrafish. Development 135: 183–192.

Yang P, Arnold SA, Habas A, et al. (2008) Ciliary neurotrophic factor mediates dopamine D2 receptor‐induced CNS neurogenesis in adult mice. Journal of Neuroscience 28: 2231–2241.

Yin VP, Thomson JM, Thummel R, et al. (2008) Fgf‐dependent depletion of microRNA‐133 promotes appendage regeneration in zebrafish. Genes & Development 22: 728–733.

Yin VP, Lepilina A, Smith A, et al. (2012) Regulation of zebrafish heart regeneration by miR‐133. Developmental Biology 365: 319–327.

Yu Y‐M, Cristofanilli M, Valiveti A, et al. (2011a) The extracellular matrix glycoprotein tenascin‐c promotes locomoter recovery after spinal cord injury in adult zebrafish. Neuroscience 183: 238–250.

Yu Y‐M, Gibbs KM, Davila J, et al. (2011b) MicroRNA miR‐133b is essential for functional recovery after spinal cord injury in adult zebrafish. European Journal of Neuroscience 33: 1587–1597.

Zhang J, Jeradi S, Strähle U, et al. (2012) Laser ablation of the sonic hedgehog‐a‐expressing cells during fin regeneration affects ray branching morphogenesis. Developmental Biology 365: 424–433.

Zhao L, Borikova AL, Ben‐Yair R, et al. (2014) Notch signaling regulates cardiomyocyte proliferation during zebrafish heart regeneration. Proceedings of the National Academy of Sciences of the United States of America 111: 1403–1408.

Zupanc GKH and Ott R (1999) Cell proliferation after lesions in the cerebellum of adult teleost fish: time course, origin, and type of new cells produced. Experimental Neurology 160: 78–87.

Further Reading

Bhatt DH, Otto SJ, Depoister B and Fetcho JR (2004) Cyclic AMP‐induced repair of zebrafish spinal circuits. Science 305: 254–258.

Goldshmit Y, Sztal TE, Jusuf PR, et al. (2012) Fgf‐dependent glial cell bridges facilitate spinal cord regeneration in zebrafish. Journal of Neuroscience 32: 7477–7492.

Gupta V and Poss KD (2012) Clonally dominant cardiomyocytes direct heart morphogenesis. Nature 484: 479–484.

Jopling C, Sleep E, Raya M, et al. (2010) Zebrafish heart regeneration occurs by cardiomyocyte dedifferentiation and proliferation. Nature 464: 606–610.

Kaslin J, Ganz J, Geffarth M, et al. (2009) Stem cells in the adult zebrafish cerebellum: initiation and maintenance of a novel stem cell niche. Journal of Neuroscience 29: 6142–6153.

Kizil C, Kyritsis N, Dudczig S, et al. (2012) Regenerative neurogenesis from neural progenitor cells requires injury‐induced expression of Gata3. Developmental Cell 23: 1230–1237.

Knopf F, Hammond C, Chekuru A, et al. (2011) Bone regeneration via dedifferentiation of osteoblasts in the zebrafish fin. Developmental Cell 20: 725–732.

Kujawski S, Lin W, Kitte F, et al. (2014) Calcineurin regulates coordinated outgrowth of zebrafish regenerating fins. Developmental Cell 28: 1–15.

Kyritsis N, Kizil C, Zocher S, et al. (2012) Acute inflammation initiates the regenerative response in the adult zebrafish brain. Science 338: 1353–1356.

Nachtrab G, Kikuchi K, Tornini VA, et al. (2013) Transcriptional components of anteroposterior positional information during zebrafish regeneration. Development 140: 3754–3764.

Neithammer P, Grabher C, Look AT, et al. (2009) A tissue‐scale gradient of hydrogen peroxide mediates rapid wound detection in zebrafish. Nature 459: 996–1000.

Reimer MM, Norris A, Ohnmacht J, et al. (2013) Dopamine from the brain promotes spinal motor neuron generation during development and adult regeneration. Developmental Cell 25 (5): 478–491.

Contact Editor close
Submit a note to the editor about this article by filling in the form below.

* Required Field

How to Cite close
Antos, Christopher L, Knopf, Franziska, and Brand, Michael(Feb 2016) Regeneration of Organs and Appendages in Zebrafish: A Window into Underlying Control Mechanisms. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1002/9780470015902.a0022101.pub2]