Electrical resonance and Ca2+ influx in the synaptic terminal of depolarizing bipolar cells from the Goldfish retina
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L. Lagnado: email@example.com
- 1Whole-cell recordings and fura-2 measurements of cytoplasmic [Ca2+] were made in depolarizing bipolar cells isolated from the retinae of goldfish. The aim was to study the voltage signal that regulates Ca2+ influx in the synaptic terminal.
- 2The current-voltage relation was linear up to about −44 mV. At this threshold, the injection of 1 pA of current triggered a maintained ‘all-or-none’ depolarization to a plateau of −34 mV, associated with a decrease in input resistance and a damped voltage oscillation with a frequency of 50–70 Hz and initial amplitude of 4–10 mV. A second frequency component of 5–10 Hz was often observed. In a minority of cells the response to current injection was transient, recovering with an undershoot.
- 3Unstimulated bipolar cells generated similar voltage signals, driven by current entering the cell through a non-specific cation conductance that continuously varied in amplitude.
- 4The threshold for activation of the Ca2+ current was −43 mV and free [Ca2+]i in the synaptic terminal rose during a depolarizing response. Simultaneous measurements of the fluorescence associated with the membrane marker FM1–43 demonstrated that these Ca2+ signals stimulated exocytosis. Regenerative depolarizations and associated rises in [Ca2+]i were blocked by inhibiting l-type Ca2+ channels with 30 μm nifedipine.
- 5Depolarization beyond −40 mV also elicited an outwardly rectifying K+ current. Blocking this current by replacing external Ca2+ with Ba2+ caused the voltage reached during a depolarizing response to increase to +10 mV.
- 6The majority of the K+ current was blocked by 100 nm charybdotoxin, indicating that it was carried by large-conductance Ca2+-activated K+ channels. A transient voltage-gated K2+current remained, which began to activate at −40 mV. High-frequency voltage oscillations were blocked by 100 nm charybdotoxin, but low-frequency oscillations remained.
- 7These results indicate that the voltage response of depolarizing bipolar cells is shaped by l-type Ca2+ channels, Ca+-activated K+ channels and voltage-dependent K+ channels. This combination of conductances regulates Ca2+ influx into the synaptic terminal and confers an electrical resonance on the bipolar cell.