Y2 receptor expression and inhibition of voltage-dependent Ca2+ influx into rod bipolar cell terminals

Neuropeptide Y (NPY) is a potent inhibitory neuropeptide expressed by amacrine cells in the rat retina. NPY modulates the release of multiple neurotransmitters in mammalian retina, yet the mechanisms mediating this regulation are not well defined. To further understand the action of NPY in the retina, Y receptor coupling to voltage-dependent Ca(2+) channels was investigated using Ca(2+) imaging with fura-2 AM to measure [Ca(2+)](i) increases in rod bipolar cell terminals. Y receptor expression was studied in rat retinal tissue with reverse transcription-polymerase chain reaction (RT-PCR). NPY inhibited the depolarization-evoked Ca(2+) influx into rod bipolar cell axon terminals and caused a dose-dependent reduction and an average maximal inhibition of 72% at 1 microM, which was reversed upon washout. K(+)-evoked Ca(2+) increases were also inhibited by the selective Y2 receptor agonists, C2-NPY and NPY(13-36), at concentrations of 1 microM, but not by the selective Y1 receptor agonist, [Leu(31)Pro(34)]NPY, selective Y4 receptor agonist, rPP, or the selective Y5 receptor agonist, [d-Trp32]-NPY. Y receptor expression was determined using RT-PCR for all known Y receptor subtypes. Y2 receptor mRNA, as well as Y1, Y4, and Y5 receptor mRNAs, are present in the rat retina. Like the rod bipolar cell, other studies in central neurons have shown that the Y2 receptor is expressed predominantly as a presynaptic receptor and that it modulates transmitter release. Together, these findings suggest that NPY activates presynaptic Y2 receptors to inhibit voltage-dependent Ca(2+) influx into rod bipolar cell terminals, and establishes one mechanism by which NPY may reduce l-glutamate release from the rod bipolar cell synapse.