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Retinal bipolar cells (BCs) compose the canonical vertical excitatory pathway that conveys photoreceptor output to inner retinal neurons. Although synaptic transmission from BC terminals is thought to rely almost exclusively on Ca
influx through voltage-gated Ca
(Ca
) channels mediating L-type currents, the molecular identity of Ca
channels in BCs is uncertain. Therefore, we combined molecular and functional analyses to determine the expression profiles of Ca
α
, β, and α
δ subunits in mouse rod bipolar (RB) cells, BCs from which the dynamics of synaptic transmission are relatively well-characterized. We found significant heterogeneity in Ca
subunit expression within the RB population from mice of either sex, and significantly, we discovered that transmission from RB synapses was mediated by Ca
influx through P/Q-type (Ca
2.1) and N-type (Ca
2.2) conductances as well as the previously-described L-type (Ca
1) and T-type (Ca
3) conductances. Furthermore, we found both Ca
1.3 and Ca
1.4 proteins located near presynaptic ribbon-type active zones in RB axon terminals, indicating that the L-type conductance is mediated by multiple Ca
1 subtypes. Similarly, Ca
3 α
, β, and α
δ subunits also appear to obey a "multisubtype" rule, i.e., we observed a combination of multiple subtypes, rather than a single subtype as previously thought, for each Ca
subunit in individual cells.
Bipolar cells (BCs) transmit photoreceptor output to inner retinal neurons. Although synaptic transmission from BC terminals is thought to rely almost exclusively on Ca
influx through L-type voltage-gated Ca
(Ca
) channels, the molecular identity of Ca
channels in BCs is uncertain. Here, we report unexpectedly high molecular diversity of Ca
subunits in BCs. Transmission from rod bipolar (RB) cell synapses can be mediated by Ca
influx through P/Q-type (Ca
2.1) and N-type (Ca
2.2) conductances as well as the previously-described L-type (Ca
1) and T-type (Ca
3) conductances. Furthermore, Ca
1, Ca
3, β, and α
δ subunits appear to obey a "multisubtype" rule, i.e., a combination of multiple subtypes for each subunit in individual cells, rather than a single subtype as previously thought.