, Volume 20, Issue 1, pp 109–125 | Cite as

Extensive growth is followed by neurodegenerative pathology in the continuously expanding adult zebrafish retina

  • Jessie Van houcke
  • Emiel Geeraerts
  • Sophie Vanhunsel
  • An Beckers
  • Lut Noterdaeme
  • Marijke Christiaens
  • Ilse Bollaerts
  • Lies De Groef
  • Lieve MoonsEmail author
Research Article


The development of effective treatments for age-related neurodegenerative diseases remains one of the biggest medical challenges today, underscoring the high need for suitable animal model systems to improve our understanding of aging and age-associated neuropathology. Zebrafish have become an indispensable complementary model organism in gerontology research, yet their growth-control properties significantly differ from those in mammals. Here, we took advantage of the clearly defined and highly conserved structure of the fish retina to study the relationship between the processes of growth and aging in the adult zebrafish central nervous system (CNS). Detailed morphological measurements reveal an early phase of extensive retinal growth, where both the addition of new cells and stretching of existent tissue drive the increase in retinal surface. Thereafter, and coinciding with a significant decline in retinal growth rate, a neurodegenerative phenotype becomes apparent,–characterized by a loss of synaptic integrity, an age-related decrease in cell density and the onset of cellular senescence. Altogether, these findings support the adult zebrafish retina as a valuable model for gerontology research and CNS disease modeling and will hopefully stimulate further research into the mechanisms of aging and age-related pathology.


Zebrafish Aging Growth Retina Neurogenesis Neurodegeneration 



Choline acetyltransferase


Ciliary marginal zone


Central nervous system


Displaced amacrine cell




Ganglion cell layer


Haematoxylin and eosin




Inner nuclear layer


Inner plexiform layer


Microtubule-associated protein 2


Nerve fiber layer


Optic nerve head


Outer nuclear layer


Outer plexiform layer


Optic tectum




Phosphate buffered saline


Proliferating cell nuclear antigen


Protein kinase C


Postsynaptic density protein 95


Photoreceptor layer


RNA binding protein mRNA processing factor


Retinal ganglion cell


Retinal pigment epithelium


Senescence-associated β-galactosidase


Sodium dodecyl sulfate


Synaptic vesicle protein 2


Tyrosine hydroxylase


Zebrafish International Resource Center



This work has been funded by research Grants from KU Leuven (BOF-OT14/00830), the Research Foundation Flanders (FWO-Vlaanderen, Belgium, G0B2315N and G053217N), and the Hercules Foundation (equipment grants AKUL/09/038 & AKUL/13/09). SV is supported by the Flemish government agency for Innovation by Science and Technology (IWT-Vlaanderen, Belgium), and LDG is a postdoctoral fellow funded by the Research foundation Flanders and KU Leuven.

Compliance with ethical standards

Conflicts of interest

The authors have no actual or potential conflicts of interest.


  1. Aggarwal P, Nag TC, Wadhwa S (2007) Age-related decrease in rod bipolar cell density of the human retina: an immunohistochemical study. J Biosci 32:293–298CrossRefGoogle Scholar
  2. Ali M (1964) Stretching of retina during growth of salmon (Salmo salar). Growth 28:83–89Google Scholar
  3. Anderton BH, Callahan L, Coleman P, Davies P, Flood D, Jicha GA, Ohm T, Weaver C (1998) Dendritic changes in Alzheimer’s disease and factors that may underlie these changes. Prog Neurobiol 55:595–609CrossRefGoogle Scholar
  4. Arenzana FJ, Santos-Ledo A, Porteros A, Aijon J, Velasco A, Lara JM, Arevalo R (2011) Characterisation of neuronal and glial populations of the visual system during zebrafish lifespan. Int J Dev Neurosci 29:441–449. CrossRefGoogle Scholar
  5. Armstrong HA, Smith CJ (2001) Growth patterns in euconodont crown enamel: implications for life history and mode-of-life reconstruction in the earliest vertebrates. Proc Biol sci R Soc 268:815–820. CrossRefGoogle Scholar
  6. Arslan-Ergul A, Adams MM (2014) Gene expression changes in aging zebrafish (Danio rerio) brains are sexually dimorphic. BMC Neurosci 15:29. CrossRefGoogle Scholar
  7. Arslan-Ergul A, Erbaba B, Karoglu ET, Halim DO, Adams MM (2016) Short-term dietary restriction in old zebrafish changes cell senescence mechanisms. Neuroscience 334:64–75. CrossRefGoogle Scholar
  8. Battista AG, Ricatti MJ, Pafundo DE, Gautier MA, Faillace MP (2009) Extracellular ADP regulates lesion-induced in vivo cell proliferation and death in the zebrafish retina. J Neurochem 111:600–613. CrossRefGoogle Scholar
  9. Bernardos RL, Barthel LK, Meyers JR, Raymond PA (2007) Late-stage neuronal progenitors in the retina are radial Muller glia that function as retinal stem cells. J Neurosci 27:7028–7040. CrossRefGoogle Scholar
  10. Bilotta J, Saszik S (2001) The zebrafish as a model visual system. Int J Dev Neurosci 19:621–629CrossRefGoogle Scholar
  11. Braak E, Braak H (1997) Alzheimer’s disease: transiently developing dendritic changes in pyramidal cells of sector CA1 of the Ammon’s horn. Acta Neuropathol 93:323–325CrossRefGoogle Scholar
  12. Bringmann A, Pannicke T, Grosche J, Francke M, Wiedemann P, Skatchkov SN, Osborne NN, Reichenbach A (2006) Muller cells in the healthy and diseased retina. Prog Retin Eye Res 25:397–424. CrossRefGoogle Scholar
  13. Campbell DS, Okamoto H (2013) Local caspase activation interacts with Slit-Robo signaling to restrict axonal arborization. J Cell Biol 203:657–672. CrossRefGoogle Scholar
  14. Centanin L, Hoeckendorf B, Wittbrodt J (2011) Fate restriction and multipotency in retinal stem cells. Cell Stem Cell 9:553–562. CrossRefGoogle Scholar
  15. Curcio CA, Millican CL, Allen KA, Kalina RE (1993) Aging of the human photoreceptor mosaic: evidence for selective vulnerability of rods in central retina. Investig Ophthalmol Vis Sci 34:3278–3296Google Scholar
  16. D’Amelio M, Cavallucci V, Middei S, Marchetti C, Pacioni S, Ferri A, Diamantini A, De Zio D, Carrara P, Battistini L, Moreno S, Bacci A, Ammassari-Teule M, Marie H, Cecconi F (2011) Caspase-3 triggers early synaptic dysfunction in a mouse model of Alzheimer’s disease. Nat Neurosci 14:69–76. CrossRefGoogle Scholar
  17. Della Santina L, Inman DM, Lupien CB, Horner PJ, Wong RO (2013) Differential progression of structural and functional alterations in distinct retinal ganglion cell types in a mouse model of glaucoma. J Neurosci 33:17444–17457. CrossRefGoogle Scholar
  18. Easter SS Jr (1992) Retinal growth in foveated teleosts: nasotemporal asymmetry keeps the fovea in temporal retina. J Neurosci 12:2381–2392CrossRefGoogle Scholar
  19. Easter SS Jr, Johns PR, Baumann LR (1977) Growth of the adult goldfish eye–I: optics. Vis Res 17:469–477CrossRefGoogle Scholar
  20. Edelmann K, Glashauser L, Sprungala S, Hesl B, Fritschle M, Ninkovic J, Godinho L, Chapouton P (2013) Increased radial glia quiescence, decreased reactivation upon injury and unaltered neuroblast behavior underlie decreased neurogenesis in the aging zebrafish telencephalon. J Comp Neurol 521:3099–3115. CrossRefGoogle Scholar
  21. Erturk A, Wang Y, Sheng M (2014) Local pruning of dendrites and spines by caspase-3-dependent and proteasome-limited mechanisms. J Neurosci 34:1672–1688. CrossRefGoogle Scholar
  22. Gestri G, Link BA, Neuhauss SC (2012) The visual system of zebrafish and its use to model human ocular diseases. Dev Neurobiol 72:302–327. CrossRefGoogle Scholar
  23. Gray DA, Woulfe J (2005) Lipofuscin and aging: a matter of toxic waste. Sci Aging Knowl Environ. Google Scholar
  24. Johns PR (1977) Growth of the adult goldfish eye. III. Source of the new retinal cells. J Comp Neurol 176:343–357. CrossRefGoogle Scholar
  25. Johns PR, Easter SS Jr (1977) Growth of the adult goldfish eye. II. Increase in retinal cell number. J Comp Neurol 176:331–341. CrossRefGoogle Scholar
  26. Kishi S (2011) The search for evolutionary developmental origins of aging in zebrafish: a novel intersection of developmental and senescence biology in the zebrafish model system. Birth Defect Res Part C 93:229–248. CrossRefGoogle Scholar
  27. Kishi S, Bayliss PE, Uchiyama J, Koshimizu E, Qi J, Nanjappa P, Imamura S, Islam A, Neuberg D, Amsterdam A, Roberts TM, Lee BY, Han JA, Im JS, Morrone A, Johung K, Goodwin EC, Kleijer WJ, DiMaio D, Hwang ES (2008) The identification of zebrafish mutants showing alterations in senescence-associated biomarkers. PLoS Genet 4:e1000152. CrossRefGoogle Scholar
  28. Kishi S, Slack BE, Uchiyama J, Zhdanova IV (2009) Zebrafish as a genetic model in biological and behavioral gerontology: where development meets aging in vertebrates–a mini-review. Gerontology 55:430–441. CrossRefGoogle Scholar
  29. Knafo S, Alonso-Nanclares L, Gonzalez-Soriano J, Merino-Serrais P, Fernaud-Espinosa I, Ferrer I, DeFelipe J (2009) Widespread changes in dendritic spines in a model of Alzheimer’s disease. Cereb Cortex 19:586–592. CrossRefGoogle Scholar
  30. Kock J-H (1982) Neuronal addition and retinal expansion during growth of the crucian carp eye. J Comp Neurol 209:264–274. CrossRefGoogle Scholar
  31. Kock J-H, Reuter T (1978) Retinal ganglion cells in the crucian carp (Carassius carassius). I. Size and number of somata in eyes of different size. J Comp Neurol 179:535–547. CrossRefGoogle Scholar
  32. Lee BY, Han JA, Im JS, Morrone A, Johung K, Goodwin EC, Kleijer WJ, DiMaio D, Hwang ES (2006) Senescence-associated beta-galactosidase is lysosomal beta-galactosidas. Aging Cell 5:187–195. CrossRefGoogle Scholar
  33. Lepanto P, Davison C, Casanova G, Badano JL, Zolessi FR (2016) Characterization of primary cilia during the differentiation of retinal ganglion cells in the zebrafish. Neural Dev 11:10. CrossRefGoogle Scholar
  34. Lieven CJ, Millet LE, Hoegger MJ, Levin LA (2007) Induction of axon and dendrite formation during early RGC-5 cell differentiation. Exp Eye Res 85:678–683. CrossRefGoogle Scholar
  35. Lopez-Otin C, Blasco MA, Partridge L, Serrano M, Kroemer G (2013) The hallmarks of aging. Cell 153:1194–1217. CrossRefGoogle Scholar
  36. Lyall A (1957) The Growth of the Trout Retina. J Cell Sci 3:101–110Google Scholar
  37. Mack AF, Fernald RD (1995) New rods move before differentiating in adult teleost retina. Dev Biol 170:136–141. CrossRefGoogle Scholar
  38. Mack AF, Sussmann C, Hirt B, Wagner HJ (2004) Displaced amacrine cells disappear from the ganglion cell layer in the central retina of adult fish during growth. Investig Ophthalmol Vis Sci 45:3749–3755. CrossRefGoogle Scholar
  39. Manuel R, Gorissen M, Stokkermans M, Zethof J, Ebbesson LO, Vis HV, Flik G, Bos RV (2015) The effects of environmental enrichment and age-related differences on inhibitory avoidance in Zebrafish (Danio rerio Hamilton). Zebrafish. Google Scholar
  40. Marcus RC, Delaney CL, Easter SS (1999) Neurogenesis in the visual system of embryonic and adult zebrafish (Danio rerio). Vis Neurosci 16:417–424CrossRefGoogle Scholar
  41. Mattson MP, Magnus T (2006) Ageing and neuronal vulnerability. Nat Rev Neurosci 7:278–294. CrossRefGoogle Scholar
  42. Matus A (1991) Microtubule-associated proteins and neuronal morphogenesis. J Cell Sci Suppl 15:61–67CrossRefGoogle Scholar
  43. Meyer RL (1978) Evidence from thymidine labeling for continuing growth of retina and tectum in juvenile goldfish. Exp Neurol 59:99–111. CrossRefGoogle Scholar
  44. Meyer MP, Trimmer JS, Gilthorpe JD, Smith SJ (2005) Characterization of zebrafish PSD-95 gene family members. J Neurobiol 63:91–105. CrossRefGoogle Scholar
  45. Morgunova G, Kolesnikov A, Klebanov A, Khokhlov A (2015) Senescence-associated β-galactosidase—A biomarker of aging, DNA damage, or cell proliferation restriction? Mosc Univ Biol Sci Bull 70:165–167CrossRefGoogle Scholar
  46. Müller H (1952) Bau und Wachstum der netzhaut des Guppy (Lebistes reticulatus). Zool Jb 63:275–324Google Scholar
  47. Nagashima M, Barthel LK, Raymond PA (2013) A self-renewing division of zebrafish Muller glial cells generates neuronal progenitors that require N-cadherin to regenerate retinal neurons. Development 140:4510–4521. CrossRefGoogle Scholar
  48. Okabe S, Shiomura Y, Hirokawa N (1989) Immunocytochemical localization of microtubule-associated proteins 1A and 2 in the rat retina. Brain Res 483:335–346CrossRefGoogle Scholar
  49. Pamela Raymond J (1981) Growth of Fish Retinas. Am Zool 21:447–458CrossRefGoogle Scholar
  50. Panda-Jonas S, Jonas JB, Jakobczyk-Zmija M (1995) Retinal photoreceptor density decreases with age. Ophthalmology 102:1853–1859CrossRefGoogle Scholar
  51. Philpott C, Donack CJ, Cousin MA, Pierret C (2012) Reducing the noise in behavioral assays: sex and age in adult zebrafish locomotion. Zebrafish 9:191–194. CrossRefGoogle Scholar
  52. Ramirez G, Alvarez A, Garcia-Abreu J, Gomes FC, Moura Neto V, Maccioni RB (1999) Regulatory roles of microtubule-associated proteins in neuronal morphogenesis. Involvement of the extracellular matrix. Braz J Med Biol Res 32:611–618CrossRefGoogle Scholar
  53. Raymond PA, Rivlin PK (1987) Germinal cells in the goldfish retina that produce rod photoreceptors. Dev Biol 122:120–138CrossRefGoogle Scholar
  54. Ruhl T, Jonas A, Seidel NI, Prinz N, Albayram O, Bilkei-Gorzo A, von der Emde G (2015) Oxidation and Cognitive Impairment in the Aging Zebrafish. Gerontology 62:47–57. CrossRefGoogle Scholar
  55. Sager JJ, Bai Q, Burton EA (2010) Transgenic zebrafish models of neurodegenerative diseases. Brain Struct Funct 214:285–302. CrossRefGoogle Scholar
  56. Samuel MA, Zhang Y, Meister M, Sanes JR (2011) Age-related alterations in neurons of the mouse retina. J Neurosci 31:16033–16044. CrossRefGoogle Scholar
  57. Stenkamp DL (2011) The rod photoreceptor lineage of teleost fish. Prog Retin Eye Res 30:395–404. CrossRefGoogle Scholar
  58. Tripathi A (2012) New cellular and molecular approaches to ageing brain. Ann Neurosci 19:177–182. CrossRefGoogle Scholar
  59. Tsai SBT, Uchiyama V, Fabian NJ, Lin MC, Bayliss PE, Neuberg DS, Zhdanova IV, Kishi S (2007) Differential effects of genotoxic stress on both concurrent body growth and gradual senescence in the adult zebrafish. Aging Cell 6:209–224. CrossRefGoogle Scholar
  60. Uylings HB, de Brabander JM (2002) Neuronal changes in normal human aging and Alzheimer’s disease. Brain Cogn 49:268–276CrossRefGoogle Scholar
  61. Van houcke J, De Groef L, Dekeyster E, Moons L (2015) The zebrafish as a gerontology model in nervous system aging, disease, and repair. Ageing Res Rev 24:358–368CrossRefGoogle Scholar
  62. Vitorino M, Jusuf PR, Maurus D, Kimura Y, Higashijima S, Harris WA (2009) Vsx2 in the zebrafish retina: restricted lineages through derepression. Neural Dev 4:14. CrossRefGoogle Scholar
  63. Wanagat J, Allison DB, Weindruch R (1999) Caloric intake and aging: mechanisms in rodents and a study in nonhuman primates. Toxicol Sci 52:35–40CrossRefGoogle Scholar
  64. Westerfield M (2000) The zebrafish book A guide for the laboratory use of zebrafish (Danio rerio), 4th edn. Univiversity of Oregon Press, EugeneGoogle Scholar
  65. Yazulla S, Studholme KM (2001) Neurochemical anatomy of the zebrafish retina as determined by immunocytochemistry. J Neurocytol 30:551–592CrossRefGoogle Scholar
  66. Yu L, Tucci V, Kishi S, Zhdanova IV (2006) Cognitive aging in zebrafish. PLoS ONE 1:e14. CrossRefGoogle Scholar
  67. Zhdanova IV, Yu L, Lopez-Patino M, Shang E, Kishi S, Guelin E (2008) Aging of the circadian system in zebrafish and the effects of melatonin on sleep and cognitive performance. Brain Res Bull 75:433–441. CrossRefGoogle Scholar
  68. Zygar CA, Lee MJ, Fernald RD (1999) Nasotemporal asymmetry during teleost retinal growth: preserving an area of specialization. J Neurobiol 41:435–442CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  1. 1.Neural Circuit Development and Regeneration Research Group, Animal Physiology and Neurobiology Section, Department of BiologyKU LeuvenLeuvenBelgium

Personalised recommendations