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Molecular & Cellular Proteomics 2:494-505, 2003.
© 2003 by The American Society for Biochemistry and Molecular Biology, Inc.

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Research

Differential Proteomic Analysis of the Mouse Retina

The Induction of Crystallin Proteins by Retinal Degeneration in the rd1 Mouse^*

Nükhet Cavusoglu^,, Danièle Thierse^¶, Saddek Mohand-Saïd^¶, Frédéric Chalmel^||, Olivier Poch^||, Alain Van-Dorsselaer, José-Alain Sahel^¶ and Thierry Léveillard^||,**

From the Laboratoire de Spectrométrie de Masse Bio-Organique, CNRS UMR 7509, Ecole Européenne de Chimie, Polyméres et Matériaux, 25 rue Becquerel, 67087 Strasbourg Cedex, France, ^¶ Laboratoire de Physiopathologie moléculaire et cellulaire de la rétine, INSERM U592, Hôpital St-Antoine, 184 rue du Faubourg St-Antoine, 75571, Paris Cedex 12, France, and ^|| Institut de Génétique et de Biologie Moléculaire et Cellulaire, 1 rue Laurent Fries, 67404 Illkirch, France

We have applied proteomic analysis to the degeneration of photoreceptors. In the rd1 mouse, a recessive mutation in the PDE6B gene leads to rapid loss of rods through apoptosis. By 5 wk postnatal, virtually all rod photoreceptors have degenerated, leaving one row of cones that degenerates secondarily. In order to assess comparative protein expression, proteins extracted from whole retina were resolved on a two-dimensional gel and identified by mass spectrometry combined with database screening. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry coupled to peptide mass fingerprinting was sufficient to identify most of the proteins, the remaining being identified with additional sequence information obtained by nano-electrospray ionization tandem mass spectrometry or liquid chromatography tandem mass spectrometry. The study revealed 212 spots, grouped into 109 different proteins. Differential analysis showed loss of proteins involved in the rod-specific phototransduction cascade, as well as induction of proteins from the crystallin family, in response to retinal degeneration. Identification of such pathways may contribute to new therapeutic approaches.

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