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In the mammalian retina, divergence of cone signals into multiple feedforward bipolar cell pathways forms the initial basis for parallel retinal circuits dedicated to specific visual functions. Here, we used patch-clamp electrophysiology, electron microscopy, and two-photon imaging of a fluorescent glutamate sensor to examine how kinetically distinct responses arise in transient versus sustained ON alpha retinal ganglion cells (ON-T and ON-S RGCs) of the mouse retina. We directly compared the visual response properties of these RGCs with their presynaptic bipolar cell partners, which we identified using 3D electron microscopy reconstruction. Different ON bipolar cell subtypes (types 5i, 6, and 7) had indistinguishable light-driven responses whereas extracellular glutamate signals around RGC dendrites and postsynaptic excitatory currents measured in ON-T and ON-S RGCs in response to the identical stimuli used to probe bipolar cells were kinetically distinct. Anatomical examination of the bipolar cell axon terminals presynaptic to ON-T and ON-S RGCs suggests that bipolar subtype-specific differences in the size of synaptic ribbon-associated vesicle pools may contribute to transient versus sustained kinetics. Our findings indicate that feedforward bipolar cell synapses are a primary point of divergence in kinetically distinct visual pathways.<\/jats:p>","DOI":"10.7554\/elife.98817","type":"journal-article","created":{"date-parts":[[2024,8,20]],"date-time":"2024-08-20T12:42:25Z","timestamp":1724157745000},"update-policy":"https:\/\/doi.org\/10.7554\/elife.98817","source":"Crossref","is-referenced-by-count":1,"title":["Cone bipolar cell synapses generate transient versus sustained signals in parallel ON pathways of the mouse retina"],"prefix":"10.7554","volume":"13","author":[{"given":"Sidney P","family":"Kuo","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00cvxb145","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Neurobiology and Biophysics, University of Washington","place":["Seattle, United States"]},{"id":[{"id":"https:\/\/ror.org\/04s5mat29","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Biology, University of 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