May 2003
Volume 44, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2003
Peptides ADNF-9 and NAP Increase Cell Survival and Neurite Outgrowth of Rat Retinal Ganglion Cells In Vitro
Author Affiliations & Notes
  • W.A. Lagreze
    Universitäts-Augenklinik, Freiburg, Germany
  • A. Pielen
    Universitäts-Augenklinik, Freiburg, Germany
  • R. Steingart
    Department of Clinical Biochemistry, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel
  • H.D. Hofmann
    Anatomy I, University Freiburg, Freiburg, Germany
  • I. Gozes
    Anatomy I, University Freiburg, Freiburg, Germany
  • M. Kirsch
    Anatomy I, University Freiburg, Freiburg, Germany
  • Footnotes
    Commercial Relationships  W.A. Lagreze, None; A. Pielen, None; R. Steingart, None; H.D. Hofmann, None; I. Gozes, None; M. Kirsch, None.
Investigative Ophthalmology & Visual Science May 2003, Vol.44, 5229. doi:
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      W.A. Lagreze, A. Pielen, R. Steingart, H.D. Hofmann, I. Gozes, M. Kirsch; Peptides ADNF-9 and NAP Increase Cell Survival and Neurite Outgrowth of Rat Retinal Ganglion Cells In Vitro . Invest. Ophthalmol. Vis. Sci. 2003;44(13):5229.

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

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Abstract

Abstract: : Purpose: Recent studies demonstrated that short peptides derived from activity-dependent neuroprotective protein are neuroprotective at femtomolar concentrations. We evaluated these findings in cultures of purified retinal ganglion cells (RGC) using two such peptides, ADNF-9 and NAP. In a second step we investigated the influence on neurite outgrowth and regeneration in retinal explants. Methods: RGCs were purified from newborn (P0 - P2) rat retina by immunopanning with antibodies against Thy1.1 and cultured in serumfree N2 medium for 2 days. RGCs were treated with ADNF-9 and NAP at concentrations ranging from 10e-18 to 10e-10 M. Survival was quantified by counting viable cells under phase-contrast microscopy and normalizing to controls. Retinal explants from postnatal (P9 - P11) rats were cultured in three-dimensional fibrin clots in serumfree medium for 3 days. Explants were treated with NAP (1µM) and ADNF-9 (1 µM). Neurite outgrowth was visualized by staining with sudan black and quantified by measuring axonal length. Results: Both peptides enhanced survival of RGCs in a dose-dependent manner. ADNF-9 showed a maximal effect at 10e-13 M with an increase of survival to 177%, CI95 [149, 204]. NAP showed a maximal effect at 5e-12 M with an increase of survival to 167%, CI95 [146,189]. In the explants, 1 µM ADNF-9 enhanced axonal outgrowth to 124%, CI95 [116, 133] and 1 µM NAP to 114%, CI95 [94, 135]. Conclusions: Both peptides, ADNF-9 and NAP, do not only increase neuronal cell survival in culture but also support regeneration and neurite outgrowth in retinal explants. These peptides deserve further attention as potential neuroprotective compounds.

Keywords: neuropeptides • neuroprotection • ganglion cells 
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