April 2014
Volume 55, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2014
Inhibitor of Apoptosis Stimulating Protein of p53 (iASPP) is required for retinal ganglion cell survival after axonal injury
Author Affiliations & Notes
  • Ariel Wilson
    Neuroscience, University of Montreal, Montreal, QC, Canada
  • Vince Chiodo
    Ophthalmology, University of Florida, Gainesville, FL
  • Sanford L Boye
    Ophthalmology, University of Florida, Gainesville, FL
  • Nicholas Brecha
    Neurobiology and Medicine, University of California Los Angeles, Los Angeles, CA
  • William W Hauswirth
    Ophthalmology, University of Florida, Gainesville, FL
  • Adriana Di Polo
    Neuroscience, University of Montreal, Montreal, QC, Canada
  • Footnotes
    Commercial Relationships Ariel Wilson, None; Vince Chiodo, None; Sanford Boye, None; Nicholas Brecha, None; William Hauswirth, None; Adriana Di Polo, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science April 2014, Vol.55, 4036. doi:
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      Ariel Wilson, Vince Chiodo, Sanford L Boye, Nicholas Brecha, William W Hauswirth, Adriana Di Polo; Inhibitor of Apoptosis Stimulating Protein of p53 (iASPP) is required for retinal ganglion cell survival after axonal injury. Invest. Ophthalmol. Vis. Sci. 2014;55(13):4036.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: p53 apoptotic activity is tightly regulated by the apoptosis-stimulating proteins of p53 (ASPP) family members: ASPP1, ASPP2 and iASPP. We previously showed that pro-apoptotic members ASPP1 and ASPP2 contribute to the p53-dependent death of retinal ganglion cells (RGC), however the role of the p53 inhibitor iASPP in the central nervous system is unknown. Here, we addressed the role of iASPP on RGC survival in a model of acute optic nerve injury (axotomy) using loss-of-function and gain-of-function experiments in vivo.

Methods: iASPP knockdown was carried out by intravitreal injection of small interference RNA (si-iASPP). Overexpression of iASPP in RGCs was achieved by intraocular delivery of a tyrosine mutant serotype 2 adeno-associated virus (AAV.iASPP). Phosphoserine immunoprecipitation was performed on retinal lysates of intact, axotomized, and iASPP overexpressing retinas. iASPP, Fas/CD95, PUMA, Noxa and Bax protein levels were examined by retinal immunohistochemistry and western blot analysis. RGC immunolabeling was performed with an RBPMS antibody. RGC densities were assessed by quantification of Brn3a-positive cells on retinal whole mounts followed by statistical analysis using one-way analysis of variance (ANOVA) followed by Bonferroni’s multiple comparison post-test.

Results: Our data demonstrate that iASPP is expressed by intact and injured RGCs, and that iASPP phosphoserine levels, which increase iASPP affinity towards p53, are significantly reduced following axotomy. We show that iASPP downregulation by siRNA exacerbates RGC death, whereas selective AAV-mediated overexpression of iASPP promotes robust RGC survival. iASPP overexpression results in an increase of iASPP phosphoserine levels following axotomy compared to retinas treated with a control virus. Analysis of p53 downstream targets demonstrate that increasing iASPP levels in RGCs leads to downregulation of pro-apoptotic PUMA and Fas/CD95.

Conclusions: Our study demonstrates a novel role for iASPP in the death of RGCs, and provides further evidence of the importance of ASPP family in CNS neuronal survival after axonal injury.

Keywords: 615 neuroprotection • 426 apoptosis/cell death • 531 ganglion cells  
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