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Originally published In Press as doi:10.1074/mcp.M600245-MCP200 on January 29, 2007.
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Molecular & Cellular Proteomics 6:895-907, 2007.
© 2007 by The American Society for Biochemistry and Molecular Biology, Inc.

Research

Elongation of Axons during Regeneration Involves Retinal Crystallin ß b2 (crybb2) *,S

Thomas Liedtke{ddagger}, Jens Christian Schwamborn§, Uwe Schröer{ddagger} and Solon Thanos{ddagger},¶,||

From the {ddagger} Department of Experimental Ophthalmology, Interdisciplinary Center of Clinical Research (IZKF), School of Medicine, University of Münster, Domagkstrasse 15, D-48149 Münster and § Abteilung für Molekularbiologie, Institut für Allgemeine Zoologie und Genetik, University of Münster, D-48149 Münster, Germany

Adult retinal ganglion cells (RGCs) can regenerate their axons in vitro. Using proteomics, we discovered that the supernatants of cultured retinas contain isoforms of crystallins with crystallin ß b2 (crybb2) being clearly up-regulated in the regenerating retina. Immunohistochemistry revealed the expression of crybb within the retina, including in filopodial protrusions and axons of RGCs. Cloning and overexpression of crybb2 in RGCs and hippocampal neurons increased axonogenesis, which in turn could be blocked with antibodies against ß-crystallin. Conditioned medium from crybb2-transfected cell cultures also supported the growth of axons. Finally real time imaging of the uptake of green fluorescent protein-tagged crybb2 fusion protein showed that this protein becomes internalized. These data are the first to show that axonal regeneration is related to crybb2 movement. The results suggest that neuronal crystallins constitute a novel class of neurite-promoting factors that likely operate through an autocrine mechanism and that they could be used in neurodegenerative diseases.

|| To whom correspondence should be addressed: Dept. of Experimental Ophthalmology, School of Medicine, University of Münster, Domagkstrasse 15, D-48149 Münster, Germany. Tel.: 49-251-8356915; Fax: 49-251-8356916; E-mail: solon{at}uni-muenster.de


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