Synaptic transmission in amphibian retinae during conditions unfavourable for calcium entry into presynaptic terminals.

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Toad (Bufo marinus) retinae were peeled from the pigment epithelium and superfused over the photoreceptor surface with a calcium-poor, cobalt-rich medium. The shape of the electroretinogram indicated that post-synaptic neurones received synaptic input. Adding the putative transmitters glutamate and N-acetylhistidine changed the shape of the electroretinogram. The change suggests that an excess of the putative transmitters blocked a component of synaptic transmission that persisted when a retina was bathed in cobalt. Salamander (Ambystomatigrinum) retinae in hemisected eye cups were superfused over their vitreal surface. Intracellular responses were recorded from photoreceptors. Reducing the calcium concentration in the superfusing medium from 1 mM to less than 10 microM slowly changed responses produced by light. The change indicates that the calcium concentration in the extracellular space surrounding photoreceptors fell to less than 100 microM. When retinae were superfused with a medium containing 1 mM-calcium, 3 mM-barium, and 10 mM-tetraethylammonium (TEA), rods produced action potentials that were later blocked by adding 1 mM-cobalt. Blocking calcium channels with cobalt and lowering the extracellular calcium concentration should together block calcium-dependent synaptic transmission. Intracellular responses were recorded from horizontal cells. After replacing external calcium with cobalt the membrane potential hyperpolarized and responses produced by light became smaller but did not entirely disappear. The responses that remained were less sensitive to light and had an altered shape. The change was reversible. Similar responses could be recorded after prolonged (30-120 min) exposure to cobalt. Electrical synapses between horizontal cells were uncoupled by adding 10 microM-forskolin to the cobalt medium. The polarity of a response could then be reversed if a cell was depolarized by injecting current. The observation of a reversal potential demonstrates that the response was produced by a conductance change. Intracellular responses were recorded from depolarizing and hyperpolarizing bipolar cells while retinae were superfused with cobalt-rich medium. After changing to a cobalt-free medium containing 1 mM-calcium, responses produced by light were slightly smaller. Large responses were recorded after superfusing with cobalt-rich, calcium-poor medium for 30-120 min. The results indicate that synaptic transmission by photoreceptors continues during conditions expected to block the entry of calcium into their presynaptic terminals.

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