Synaptic localization of P2X7 receptors in the rat retina

The distribution of P2X(7) receptor (P2X(7)R) subunits was studied in the rat retina using a subunit-specific antiserum. Punctate immunofluorescence was observed in the inner and outer plexiform layers. Double labeling of P2X(7) and the horizontal cell marker, calbindin, revealed extensive colocalization in the outer plexiform layer (OPL). Significant colocalization of P2X(7)R and kinesin, a marker of photoreceptor ribbons, was also observed, indicating that this receptor may be expressed at photoreceptor terminals. Furthermore, another band of P2X(7)R puncta was identified below the level of the photoreceptor terminals, adjacent to the inner nuclear layer (INL). This band of P2X(7)R puncta colocalized with the active-zone protein, bassoon, suggesting that "synapse-like" structures exist outside photoreceptor terminals. Preembedding immunoelectron microscopy demonstrated P2X(7)R labeling of photoreceptor terminals adjacent to ribbons. In addition, some horizontal cell dendrites and putative "desmosome-like" junctions below cone pedicles were labeled. In the inner plexiform layer (IPL), P2X(7)R puncta were observed surrounding terminals immunoreactive for protein kinase C-alpha, a marker of rod bipolar cells. Double labeling with bassoon in the IPL revealed extensive colocalization, indicating that P2X(7)R is likely to be found at conventional cell synapses. This finding was confirmed at the ultrastructural level: only processes presynaptic to rod bipolar cells were found to be labeled for the P2X(7)R, as well as other conventional synapses. These findings suggest that purines play a significant role in neurotransmission within the retina, and may modulate both photoreceptor and rod bipolar cell responses.