Deletion of the p85alpha regulatory subunit of phosphoinositide 3-kinase in cone photoreceptor cells results in cone photoreceptor degeneration

Invest Ophthalmol Vis Sci. 2011 Jun 1;52(6):3775-83. doi: 10.1167/iovs.10-7139.

Abstract

Purpose: Downregulation of the retinal insulin/mTOR pathway in mouse models of retinitis pigmentosa is linked to cone cell death, which can be delayed by systemic administration of insulin. A classic survival kinase linking extracellular trophic/growth factors with intracellular antiapoptotic pathways is phosphoinositide 3-kinase (PI3K), which the authors have shown to protect rod photoreceptors from stress-induced cell death. The role of PI3K in cones was studied by conditional deletion of its p85α regulatory subunit.

Methods: Mice expressing Cre recombinase in cones were bred to mice with a floxed pi3k gene encoding the p85α regulatory subunit of the PI3K and were back-crossed to ultimately generate offspring with cone-specific p85α knockout (cKO). Cre expression and cone-specific localization were confirmed by Western blot analysis and immunohistochemistry (IHC), respectively. Cone structural integrity was determined by IHC using peanut agglutinin and an M-opsin-specific antibody. Electroretinography (ERG) was used to assess rod and cone photoreceptor function. Retinal structure was examined by light and electron microscopy.

Results: An age-related cone degeneration was found in cKO mice, evidenced by a reduction in photopic ERG amplitudes and loss of cone cells. By 12 months of age, approximately 78% of cones had died, and progressive disorganization of synaptic ultrastructure was noted in surviving cone terminals in cKO retinas. Rod viability was unaffected in p85α cKO mice.

Conclusions: The present study suggests that PI3K signaling pathway is essential for cone survival in the mouse retina.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Arrestin / metabolism
  • Blotting, Western
  • Cell Line
  • Cell Survival
  • Class Ia Phosphatidylinositol 3-Kinase / physiology*
  • Electroretinography
  • Female
  • Gene Deletion
  • Immunohistochemistry
  • Integrases
  • Male
  • Mice
  • Mice, Knockout
  • Mice, Transgenic
  • Microscopy, Fluorescence
  • Opsins / metabolism
  • Retinal Cone Photoreceptor Cells / enzymology*
  • Retinal Cone Photoreceptor Cells / ultrastructure
  • Retinal Degeneration / enzymology*
  • Retinal Degeneration / pathology

Substances

  • Arrestin
  • Opsins
  • Class Ia Phosphatidylinositol 3-Kinase
  • Cre recombinase
  • Integrases