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The Journal of Neuroscience, May 15, 1998, 18(10):3715-3724

Calcium Currents and Calcium Signaling in Rod Bipolar Cells of Rat Retinal Slices

Dario A. Protti and Isabel Llano

Arbeitsgruppe Zelluläre Neurobiologie, Max-Planck-Institut für biophysikalische Chemie, 37070 Göttingen, Germany

Combined electrophysiological and imaging techniques were used to study calcium currents (I_Ca) and their sites of origin at rod bipolar cells in rat retinal slices. We report here for the first time the successful whole-cell patch-clamp recording from presynaptic boutons that were compared with somatic recordings. TTX-resistant inward currents were elicited in response to depolarization. The kinetic and pharmacological properties of I_Ca were very similar for recordings obtained from the soma and the presynaptic terminals. I_Ca activated maximally between 30 and 20 mV was enhanced by Bay K 8644 and was blocked by isradipine and nifedipine. Peak amplitude and time to peak were 31.3 ± 1.2 pA and 3.2 ± 0.2 msec with somatic recordings (n = 54), whereas the corresponding values were 31.6 ± 6.1 pA and 3.2 ± 0.7 msec in recordings obtained directly from terminals (n = 6). I_Ca showed little inactivation during sustained depolarizations. No T-type I_Ca was observed with depolarizations from 90 mV. Concomitant with Ca²⁺ entry, depolarization induced the appearance of transient outward currents that resembled IPSCs and were blocked by GABA and glycine receptor antagonists, suggesting that they arise from activation of amacrine feedback synapses. Upon depolarization, intracellular Ca²⁺ ([Ca²⁺]_i) rises were restricted to the presynaptic terminals with no somatic or axonal changes and were linearly dependent on pulse duration when using a low-affinity Ca²⁺ indicator. In cone bipolar cells, I_Ca inactivated markedly, and [Ca²⁺]_i rises occurred in the axon, as well as in the presynaptic terminals.

Key words: retina; bipolar cells; calcium channels; presynaptic terminal; calcium imaging; CNS slices; reciprocal synapses

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Copyright © 1998 Society for Neuroscience  0270-6474/98/18103715-10$05.00/0

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