PUBLICATION

Regeneration of inner retinal neurons after intravitreal injection of ouabain in zebrafish

Authors
Fimbel, S.M., Montgomery, J.E., Burket, C.T., and Hyde, D.R.
ID
ZDB-PUB-070303-16
Date
2007
Source
The Journal of neuroscience : the official journal of the Society for Neuroscience   27(7): 1712-1724 (Journal)
Registered Authors
Burket, Christopher, Fimbel, Shane, Hyde, David R., Montgomery, Jacob
Keywords
retinal degeneration, retinal regeneration, retinal apoptosis, retinal progenitor, Müller glia, zebrafish
MeSH Terms
  • Analysis of Variance
  • Animals
  • Animals, Genetically Modified
  • Basic Helix-Loop-Helix Transcription Factors/genetics
  • Blotting, Western/methods
  • Dose-Response Relationship, Drug
  • ELAV Proteins/metabolism
  • ELAV-Like Protein 3
  • Enzyme Inhibitors/toxicity*
  • Glial Fibrillary Acidic Protein/metabolism
  • Green Fluorescent Proteins/genetics
  • Immunohistochemistry/methods
  • In Situ Nick-End Labeling/methods
  • Nerve Tissue Proteins/genetics
  • Neuroglia/metabolism
  • Neuroglia/pathology
  • Ouabain/toxicity*
  • Proliferating Cell Nuclear Antigen/metabolism
  • Regeneration/physiology*
  • Retina/pathology*
  • Retinal Ganglion Cells/drug effects*
  • Time Factors
  • Zebrafish
  • Zebrafish Proteins/genetics
  • Zebrafish Proteins/metabolism
PubMed
17301179 Full text @ J. Neurosci.
Abstract
We examined the regenerative capacity of the adult zebrafish retina by intravitreal injection of a low ouabain concentration to rapidly damage the ganglion cell layer (GCL) and inner nuclear layer (INL) with minimal photoreceptor cell damage. By 24 h after ouabain injection, maximal numbers of terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end labeling (TUNEL)-positive cells were detected in the INL and GCL, with low numbers of TUNEL-positive cells in the outer nuclear layer. Immunolabeling revealed that approximately 85% of the HuC/D-positive amacrine and ganglion cells were lost by 7 d post-ouabain injection (dpi). This ganglion cell loss was consistent with the small, but statistically significant, decrease in the optic nerve diameter. The regeneration response began within 1 dpi with increased proliferating cell nuclear antigen (PCNA) expression in both the INL and GCL. By 3 dpi, PCNA expression is primarily restricted to the Muller glia. By 5 dpi, most of the PCNA expression was localized to neuronal progenitors expressing the olig2:egfp transgene rather than the Muller glia. By 7 dpi, the neuronal progenitors began committing to the ganglion cell fate based on the coexpression of the atoh7:EGFP transgene and the zn5 antigen. The regeneration of ganglion and amacrine cells continued until 60 dpi, when they reached 75% of their uninjected control number. This demonstrates that inner retinal damage, without extensive photoreceptor damage, is sufficient to induce a regeneration response that is marked by the Muller glial cells reentering the cell cycle to produce neuronal progenitor cells that regenerate INL and ganglion cells in the zebrafish retina.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping