PUBLICATION

The inhibitor of phagocytosis, O-phospho-L-serine, suppresses Müller glia proliferation and cone cell regeneration in the light-damaged zebrafish retina

Authors
Bailey, T.J., Fossum, S.L., Fimbel, S.M., Montgomery, J.E., and Hyde, D.R.
ID
ZDB-PUB-100811-14
Date
2010
Source
Experimental Eye Research   91(5): 601-612 (Journal)
Registered Authors
Bailey, Travis, Fimbel, Shane, Hyde, David R., Montgomery, Jacob
Keywords
regeneration, Müller glia, retina, phosphatidylserine, type III metabotropic glutamate receptor, phagocytosis
MeSH Terms
  • Animals
  • Animals, Genetically Modified
  • Apoptosis
  • Cell Proliferation/drug effects*
  • Fluorescent Antibody Technique, Indirect
  • In Situ Nick-End Labeling
  • Light/adverse effects*
  • Microscopy, Confocal
  • Neuroglia/pathology*
  • Phagocytosis/drug effects
  • Phagocytosis/physiology*
  • Phosphoserine/analogs & derivatives*
  • Phosphoserine/pharmacology
  • Radiation Injuries, Experimental/metabolism
  • Radiation Injuries, Experimental/pathology*
  • Regeneration/drug effects*
  • Retina/radiation effects
  • Retinal Cone Photoreceptor Cells/physiology*
  • Retinal Degeneration/metabolism
  • Retinal Degeneration/pathology
  • Retinal Rod Photoreceptor Cells/pathology
  • Reverse Transcriptase Polymerase Chain Reaction
  • Zebrafish
PubMed
20696157 Full text @ Exp. Eye. Res.
Abstract
The damaged zebrafish retina replaces lost neurons through a regenerative response that initiates with the asymmetric cell division of Müller glia to produce neuronal progenitor cells that proliferate and migrate to the damaged retinal layer, where they differentiate into the lost neuronal cell types. Because Müller glia are known to phagocytose apoptotic retinal cells during development, we tested if Müller glia engulfed apoptotic rod cell bodies in light-damaged retinas. After 24hours of constant intense light, damaged retinas revealed both a strong nuclear TUNEL signal in photoreceptors and a weak cytoplasmic TUNEL signal in Müller glia, although Müller glial apoptosis is not observed in the light-damaged retina. Light damage of a rod-specific transgenic reporter line, Tg(XlRho:EGFP)(fl1), resulted in some Müller glia containing both TUNEL signal and EGFP, which indicated that this subset of Müller glia engulfed apoptotic photoreceptor cell bodies. To determine if phagocytosis induced the Müller glial proliferative response in the light-damaged retina, we utilized O-phospho-L-serine (L-SOP), a molecule that mimics the phosphatidylserine head group and partially blocks microglial phagocytosis of apoptotic cells. Intravitreal injection of L-SOP immediately prior to beginning constant intense light treatment: i) did not significantly reduce light-induced photoreceptor cell death, ii) significantly reduced the number of PCNA-positive Müller glia, and iii) significantly reduced the number of cone photoreceptors in the regenerated retina relative to control retinas. Because L-SOP is also a specific group III metabotropic glutamate receptor (mGluR) agonist, we also tested if the more potent specific group III agonist, L-2-amino-4-phosphonobutyrate (L-AP4), the specific group III antagonist (RS)-alpha-Methylserine-O-phosphate (MSOP) or the specific group I antagonist, L-2-amino-3-phophonopropanoic acid (L-AP3) affected Müller glial proliferation. We found no changes with any of these factors compared to control retinas, revealing that metabotropic glutamate receptors were not necessary in the Müller glia proliferative response. Furthermore, ascl1a and stat3 expression were unaffected in either the L-SOP or MSOP-injected retinas relative to controls, suggesting L-SOP disrupts Müller glia proliferation subsequent to or in parallel with ascl1a and stat3 activation. This implies that at least one signaling mechanism, in addition to the process disrupted by L-SOP, is required to activate Müller glia proliferation in the light-damaged retina.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping