TY - JOUR
T1 - Molecular Mechanisms Regulating Ocular Apoptosis in Zebrafish gdf6a Mutants
AU - Pant, Sameer D.
AU - March, Lindsey D.
AU - Famulski, Jakub K.
AU - French, Curtis R.
AU - Lehmann, Ordan J.
AU - Waskiewicz, Andrew J.
PY - 2013
Y1 - 2013
N2 - 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, nonfunctional 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. Pharmacologic inhibition (using SB203580) and IHC were used to investigate the role of p38 mitogen activated protein (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 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 pharmacomodulation in improvement of visual function needs to be further examined.
AB - 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, nonfunctional 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. Pharmacologic inhibition (using SB203580) and IHC were used to investigate the role of p38 mitogen activated protein (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 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 pharmacomodulation in improvement of visual function needs to be further examined.
U2 - 10.1167/iovs.12-11315
DO - 10.1167/iovs.12-11315
M3 - Journal article
C2 - 23847306
SN - 0146-0404
VL - 54
SP - 5871
EP - 5879
JO - Investigative Ophthalmology & Visual Science
JF - Investigative Ophthalmology & Visual Science
IS - 8
ER -