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  • Immunotoxin-induced ablation of melanopsin retinal ganglion cells in a non-murine mammalian model.

Immunotoxin-induced ablation of melanopsin retinal ganglion cells in a non-murine mammalian model.

The Journal of comparative neurology (2009-07-04)
Elizabeth S Ingham, Emine Günhan, Patrick M Fuller, Charles A Fuller
ABSTRACT

In mammals, non-image-forming visual functions, including circadian photoentrainment and the pupillary light reflex, are thought to be mediated by the combination of rods, cones, and the melanopsin-expressing intrinsically photosensitive retinal ganglion cells (ipRGCs). Although several genetic models have been developed to clarify the individual roles of the rod, cone, and ipRGC systems in mediating non-image visual function, assessing the in vivo role(s) of the ipRGCs has been complicated by the possibility of ontogenetic issues in these genetically modified animal models. In the present study, we describe the development and validation of an immunotoxin that specifically targets the ipRGC population in the mature mammalian retina. This ipRGC immunotoxin, consisting of saporin conjugated to a melanopsin polyclonal antibody, was evaluated with respect to its effectiveness and specificity in depleting the ipRGC population in the fully developed rat retina. The results showed that the ipRGC toxin rapidly and permanently depleted approximately 70% of the ipRGC population, without inducing appreciable changes in the cell number or morphology of any of the non-melanopsin-containing retinal cell populations investigated. These findings suggest that the newly developed ipRGC immunotoxin provides a potent method for achieving relatively rapid, permanent, and selective depletion of the ipRGC population in a non-murine model system. The development of this ipRGC-ablation method is the next step in elucidating the role of ipRGCs in mediating non-visual and circadian light-resetting responses in a wide range of non-murine mammalian models.

MATERIALS
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Product Description

Sigma-Aldrich
Saporin from Saponaria officinalis seeds, lyophilized powder
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