May 2004
Volume 45, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2004
Proteomic analysis of retinas from glaucomatous eyes regenerating axons in organ culture
Author Affiliations & Notes
  • J. Lasseck
    Experimental Ophthalmology, Univ Muenster Eye Inf, Muenster, Germany
  • K. Rose
    Experimental Ophthalmology, Univ Muenster Eye Inf, Muenster, Germany
  • S. Koenig
    Integrated Functional Genomics (IZKF), Muenster, Germany
  • R. Naskar
    Experimental Ophthalmology, Univ Muenster Eye Inf, Muenster, Germany
  • S. Thanos
    Experimental Ophthalmology, Univ Muenster Eye Inf, Muenster, Germany
  • Footnotes
    Commercial Relationships  J. Lasseck, None; K. Rose, None; S. Koenig, None; R. Naskar, None; S. Thanos, None.
  • Footnotes
    Support  DFG Na 425/1–1
Investigative Ophthalmology & Visual Science May 2004, Vol.45, 894. doi:
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      J. Lasseck, K. Rose, S. Koenig, R. Naskar, S. Thanos; Proteomic analysis of retinas from glaucomatous eyes regenerating axons in organ culture . Invest. Ophthalmol. Vis. Sci. 2004;45(13):894.

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

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Abstract

Abstract: : Purpose: To examine whether increased intraocular pressure (IOP) induces expression of molecules associated with regeneration of retinal ganglion cell (RGCs) axons in a rat model (GL–strain) suffering from hereditary glaucoma (buphthalmos). Methods: In the GL–rat strain elevated IOP and buphthalmic phenotype is established after the 6th week of life to continually increase throughout life. The glaucoma–induced decrease of the ganglion cell population was examined by retrograde labeling of the cells with the fluorescent carbocyanine dye 4Di–10 ASP injected into the midbrain. The regenerative potential of RGCs axons exposed to elevated IOP was examined in organ culture and compared to that of acutely axotomized RGCs and RGCs without elevated IOP. Upon regeneration of axons, the retinal tissue was harvested and used for proteomic analysis including 2D–electrophoresis, MALDI–MS sequencing, immunohistochemistry and Western blots. Results: The IOP of phenotypically buphthalmic eyes increased significantly with age whereas the IOP in the non–buphthalmic control eyes remained stable when measured with the Tonopen–XL. The number of RGCs decreased drastically in a uniform manner across all retinal quadrants and eccentricity in glaucomatous eyes. The number of regenerated axons from retinal explants was three times higher than the number of axons from controls. Among the proteins expressed within the glaucomatous retina, GAP–43 upregulation showed that glaucoma induces GAP–43. MALDI–MS–assisted peptide mapping of the regenerating retina revealed that some kinases with a as yet undefined role were differentially expressed within the regenerating retina obtained from glaucomatous eyes. Conclusion: We have presented data showing that retina obtained from eyes with chronically elevated IOP can regenerate axons. The data suggest that RGCs exposed to IOP were primed to express growth associated proteins which trigger the ability to regenerate axons.

Keywords: intraocular pressure • regeneration • ganglion cells 
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