May 2004
Volume 45, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2004
Electron microscopic localization of PDE , ß and in frog retina
Author Affiliations & Notes
  • Y. Nishizawa
    Anatomy and Cell Biology, Nagoya University, Nagoya, Japan
  • A. Yamazaki
    Kresge Eye Institute, Detroit, MI
  • J. Usukura
    Anatomy and Cell Biology, Nagoya University, Nagoya, Japan
  • Footnotes
    Commercial Relationships  Y. Nishizawa, None; A. Yamazaki, None; J. Usukura, None.
  • Footnotes
    Support  NON
Investigative Ophthalmology & Visual Science May 2004, Vol.45, 5353. doi:
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      Y. Nishizawa, A. Yamazaki, J. Usukura; Electron microscopic localization of PDE , ß and in frog retina . Invest. Ophthalmol. Vis. Sci. 2004;45(13):5353.

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Abstract

Abstract: : Purpose: We investigate precise localization of PDE α, ß and δ in the retina preserved close to the native state by rapid freezing. Methods: Retinas were isolated from dark–adapted frog, and immediately frozen quickly by plunging onto copper block cooled with liquid helium. Frozen retinas were freeze–substituted with acetone containing 1% tannic acid at –80°C. After substitution, retinas were embedded in Lowicryl K4M or LR White. For immunocytochemistry, we employed polyclonal antibodies raised against the peptide of PDE α, ß and δ. Results: Immuno–reactivity of antibodies to the frog retina was examined by western blot. Three antibodies recognized respectively well. Anti–PDE δ antibodies binding sites were found specifically in both outer and inner segments. In outer segment, especially, PDE δ was localized rich in basal region. PDE α and ß showed similar distribution in rod outer segments. It should be noted that PDE α was found frequently along the incisures. Conclusions: PDE δ as well as PDE α, ß was localized in rods and cones outer segment, but rich in the basal region. In contrast, PDE α was found frequently along the incisures.

Keywords: retina • immunohistochemistry • microscopy: electron microscopy 
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