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1 The Vollum Institute, OHSU, Portland, Oregon, United States
2 Dept. of Neurobiology, SUNY at Stony Brook, Portland, Oregon, United States
3 Stony Brook, New York, United States; Dept. of Neurobiology, SUNY at Stony Brook, Portland, Oregon, United States
4 Portland, Oregon, United States; The Vollum Institute, OHSU, Portland, Oregon, United States
* To whom correspondence should be addressed. E-mail: vongersd{at}ohsu.edu.
The number and morphology of synaptic ribbons at photoreceptor and bipolar cell terminals has been reported to change on a circadian cycle. Here, we sought to determine whether this phenomenon exists at goldfish Mb-type bipolar cell terminals with the aim of exploring the role of ribbons in transmitter release. We examined the physiology and ultrastructure of this terminal around two time points: midday and midnight. Nystatin perforated-patch recordings of membrane capacitance (Cm) revealed that synaptic vesicle exocytosis evoked by short depolarizations was reduced at night, even though Ca2+ currents were larger. The efficiency of exocytosis (measured as the 
Cm jump per total Ca2+ charge influx) was thus significantly lower at night. The paired pulse ratio remained unchanged, however, suggesting that release probability was not altered. Hence, the decreased exocytosis likely reflects a smaller readily releasable vesicle pool at night. Electron microscopy of single sections from intact retinas averaged 65% fewer ribbons at night. Interestingly, the number of active zones did not change from day to night, only the probability of finding a ribbon at an active zone. Additionally, synaptic vesicle halos surrounding the ribbons were more completely filled at night when these ON-type bipolar cells are more hyperpolarized. There was no change, however, in the physical dimensions of synaptic ribbons from day to night. These results suggest that the size of the readily releasable vesicle pool and the efficiency of exocytosis are reduced at night when fewer ribbons are present at bipolar cell terminal active zones.
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