Inhibitory control of synaptic activity in goldfish Mb bipolar cell terminals visualized by FM1-43

Abstract

To investigate the physiology and plasticity of mixed rod–cone ON-bipolar cells (Mb) in the goldfish retina, we established a slice preparation which allows us to optically monitor the synaptic activity of bipolar cell axon terminals. We used the styryl dye FM1-43 which is incorporated into active axon terminals due to synaptic vesicle cycling and thus reflects synaptic activity. Different activity states of the axon terminals were revealed when slices prepared from light-adapted retinae were incubated in the presence of FM1-43 under various conditions. Depolarizing high K⁺ Ringer (50 mM) and the gamma-butyric acid (GABA) antagonist bicuculline (100 μM) resulted in more than two-fold increase in the number of stained terminals compared to slices stained in normal Ringer. In contrast, GABA treatment (0.5 mM) reduced the frequency of stained terminals. Thus, in light-adapted retinal slices the synaptic activity of Mb axon terminals can be modulated towards higher and lower activity states. The fact that the GABA antagonist bicuculline had similar effects as stimulatory high K⁺ Ringer suggests that inhibitory control is an important component in the regulation of synaptic activity and transmitter release in Mb terminals.