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The RNA binding protein RBPMS is a selective marker of ganglion cells in the mammalian retina
Authors:Allen R. Rodriguez  Luis Pérez de Sevilla Müller  Nicholas C. Brecha
Affiliation:1. Department of Neurobiology, David Geffen School of Medicine at Los Angeles, University of California at Los Angeles, Los Angeles, California;2. Department of Medicine, David Geffen School of Medicine at Los Angeles, University of California at Los Angeles, Los Angeles, California;3. Jules Stein Eye Institute, David Geffen School of Medicine at Los Angeles, University of California at Los Angeles, Los Angeles, California;4. CURE Digestive Diseases Research Center, David Geffen School of Medicine at Los Angeles, University of California at Los Angeles, Los Angeles, California;5. Veterans Administration Greater Los Angeles Health System, Los Angeles, California
Abstract:There are few neurochemical markers that reliably identify retinal ganglion cells (RGCs), which are a heterogeneous population of cells that integrate and transmit the visual signal from the retina to the central visual nuclei. We have developed and characterized a new set of affinity‐purified guinea pig and rabbit antibodies against RNA‐binding protein with multiple splicing (RBPMS). On western blots these antibodies recognize a single band at ?24 kDa, corresponding to RBPMS, and they strongly label RGC and displaced RGC (dRGC) somata in mouse, rat, guinea pig, rabbit, and monkey retina. RBPMS‐immunoreactive cells and RGCs identified by other techniques have a similar range of somal diameters and areas. The density of RBPMS cells in mouse and rat retina is comparable to earlier semiquantitative estimates of RGCs. RBPMS is mainly expressed in medium and large DAPI‐, DRAQ5‐, NeuroTrace‐ and NeuN‐stained cells in the ganglion cell layer (GCL), and RBPMS is not expressed in syntaxin (HPC‐1)‐immunoreactive cells in the inner nuclear layer (INL) and GCL, consistent with their identity as RGCs, and not displaced amacrine cells. In mouse and rat retina, most RBPMS cells are lost following optic nerve crush or transection at 3 weeks, and all Brn3a‐, SMI‐32‐, and melanopsin‐immunoreactive RGCs also express RBPMS immunoreactivity. RBPMS immunoreactivity is localized to cyan fluorescent protein (CFP)‐fluorescent RGCs in the B6.Cg‐Tg(Thy1‐CFP)23Jrs/J mouse line. These findings show that antibodies against RBPMS are robust reagents that exclusively identify RGCs and dRGCs in multiple mammalian species, and they will be especially useful for quantification of RGCs. J. Comp. Neurol. 522:1411–1443, 2014. © 2013 Wiley Periodicals, Inc.
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