The ribbon-associated protein C-terminal-binding protein 1 is not essential for the structure and function of retinal ribbon synapses

Overall the results suggest that CtBP1 protein is not essential for the formation of functional ribbon synapses in the retina.

Confocal microscopy, electrophysiology, and electron microscopy were used to examine the structure and function of ribbon synapses in the retina and in isolated bipolar neurons from CtBP1 null mice compared with their wild-type littermates.

Synaptic ribbons are organelles found at presynaptic active zones of sensory neurons that generate sustained graded electrical signals in response to stimuli, including retinal photoreceptor cells and bipolar neurons. RIBEYE is the major and specific protein constituent of ribbons; however, over the past decade an increasing number of other proteins have been identified at ribbon active zones, including C-terminal-binding protein 1 (CtBP1; a regulator of transcription and membrane trafficking that might bind to the B domain of RIBEYE). The appearance of CtBP1 together with RIBEYE suggests that it may contribute to ribbon function, but the possible role of CtBP1 at ribbon synapses has not yet been examined. Using CtBP1-knockout mice, we tested for functional effects of absence of CtBP1 protein.

Expression of ribbons appeared to be normal in CtBP1 null mouse retina as revealed by immunofluorescence with an antibody to the B domain of RIBEYE and by binding studies using a fluorescent peptide that binds to RIBEYE in ribbons of living bipolar cells. Electron microscopy also showed grossly normal pre- and postsynaptic organization of ribbon synapses in both photoreceptors and bipolar cells. Synaptic vesicles were normal in size, but the overall density of reserve vesicles was reduced by ~20% in the cytoplasm of CtBP1 null ribbon synaptic terminals. However, the reduced vesicle density did not detectably alter synaptic function of bipolar neurons as revealed by activity-dependent loading of synaptic vesicles with FM4-64, presynaptic calcium current, capacitance measurements of synaptic exocytosis, and destaining of FM dye upon stimulation. Conclusions: Overall the results suggest that CtBP1 protein is not essential for the formation of functional ribbon synapses in the retina.