Expression of mRNA for glutamate receptor subunits distinguishes the major classes of retinal neurons, but is less specific for individual cell types.
OBJECTIVE
To investigate the expression of ionotropic glutamate receptor subunits by retinal neurons, to assess the extent to which different functional types of retinal neurons are characterized by the expression of the receptor subtypes.


METHODS
Rod photoreceptor cells and bipolar cells were identified in retina dissociates. Amacrine cells were identified in dissociates from transgenic mice or by staining with an antibody against the extracellular carbohydrate epitope CD15. Ganglion cells were identified by retrograde axonal transport of FITC-dextran or by green fluorescent protein (GFP) fluorescence in a transgenic strain. We examined the receptors simultaneously using non-quantitative single-cell reverse transcriptase polymerase chain reaction for GluR1-R4 (alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) receptors), GluR5-R7, and KA1 and 2 (kainate receptors), delta1 and delta2 subunits, and the N-methyl-D-aspartate (NMDA) receptor subunits NR1, 2a-d, and 3a.


RESULTS
The expression of glutamate receptors on bipolar cells and rod photoreceptors was limited: Neither expressed functional NMDA receptors, and rods were also negative for AMPA receptors. The sample of ganglion cells included examples of many ganglion cell types; these were distinguished morphologically using quantitative parameters defined in a previous cluster analysis. All types of ionotropic glutamate receptors were found to be expressed on ganglion cells. The iGluR subunits GluR4, KA2, delta1, and NR1 were expressed on almost all ganglion cells examined.


CONCLUSIONS
Despite the heterogeneity of ganglion cell types, differences among them in this PCR-based method were minor. Thus, retinal interneurons are characterized by expression of distinctive glutamate receptor types, but functional differences among ganglion cells seem to be reflected instead in the amounts as well as spatial distributions of a widely expressed group of receptors.
Jakobs
Tatjana C
TC
Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114, USA. Jakobs@helix.mgh.harvard.edu
Ben
Yixin
Y
Masland
Richard H
RH
D000375:Aging / Q000378:metabolism; D025042:Amacrine Cells / Q000378:metabolism; D000818:Animals; D000831:Animals, Newborn; D016000:Cluster Analysis; D003663:Decision Trees; D051379:Mice; D008810:Mice, Inbred C57BL; D009474:Neurons / Q000145:classification* / Q000378:metabolism*; D020033:Protein Isoforms / Q000235:genetics; D012333:RNA, Messenger / Q000378:metabolism*; D018091:Receptors, AMPA / Q000235:genetics; D017470:Receptors, Glutamate / Q000235:genetics*; D018092:Kainic Acid Receptors / Q000235:genetics; D016194:Receptors, N-Methyl-D-Aspartate / Q000235:genetics; D012160:Retina / Q000166:cytology* / Q000378:metabolism*; D051245:Retinal Bipolar Cells / Q000378:metabolism; D012165:Retinal Ganglion Cells / Q000378:metabolism; D017948:Retinal Rod Photoreceptor Cells / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
D020033:Protein Isoforms; D012333:RNA, Messenger; D018091:Receptors, AMPA; D017470:Receptors, Glutamate; D018092:Kainic Acid Receptors; D016194:Receptors, N-Methyl-D-Aspartate