ZFIN ID: ZDB-PUB-130722-5
Molecular mechanisms regulating ocular apoptosis in zebrafish gdf6a mutants
Pant, S.D., March, L.D., Famulski, J.K., French, C.R., Lehmann, O.J., and Waskiewicz, A.J.
Date: 2013
Source: Investigative ophthalmology & visual science   54(8): 5871-5879 (Journal)
Registered Authors: Lehmann, Ordan J., Waskiewicz, Andrew
Keywords: Gdf6, apoptosis, p38 MAP kinase, Bax, zebrafish
MeSH Terms:
  • Animals
  • Apoptosis/physiology*
  • Caspase 3/metabolism
  • Growth Differentiation Factor 6/genetics
  • Growth Differentiation Factor 6/physiology*
  • Immunohistochemistry
  • Retina/metabolism
  • Retina/physiology*
  • Signal Transduction/physiology
  • Smad1 Protein/metabolism
  • Zebrafish
  • Zebrafish Proteins/genetics
  • Zebrafish Proteins/physiology*
  • p38 Mitogen-Activated Protein Kinases/physiology
PubMed: 23847306 Full text @ Invest. Ophthalmol. Vis. Sci.

Purpose: To characterize the molecular mechanisms underlying retinal apoptosis induced by loss of Gdf6, a TGFβ ligand. Methods: The role of Gdf6 in regulating apoptosis was studied using a zebrafish gdf6a-/- mutant, which encodes a truncated non-functional protein. To investigate whether intrinsic or extrinsic apoptotic mechanisms were involved, morpholino antisense oligonucleotides targeting baxa, baxb and p53 were employed. Caspase-3 immunohistochemistry (IHC) was performed to assay apoptosis. Pharmacological inhibition (using SB203580) and IHC were used to investigate the role of p38 MAP kinase activation in gdf6a-/- induced apoptosis. To assess the role of Gdf6a in transcriptional regulation of TGFβ signal transducers, in situ Hybridization (ISH) was performed using probes to smad1, 5, 7 and 8. Results: Results indicate maximal ocular apoptosis occurs 28 hours post fertilization (hpf) in gdf6a-/- mutants that is mediated independently of p53 by intrinsic mechanisms involving Bax proteins. Also, gdf6a-/- mutants exhibit markedly increased p38 MAP kinase activation that can be inhibited to significantly reduce retinal apoptosis. A reduction in retinal smad1 and 8 expression was also noted in gdf6a-/- mutants. Conclusions: gdf6a-/- induced apoptosis is characterized by the involvement of intrinsic apoptotic pathways, p38 MAP kinases and dysregulated smad expression. Modulation of key mediators can inhibit retinal apoptosis offering potential avenues of therapy. However, the efficacy of pharmaco-modulation in improvement of visual function needs to be further examined.