Sustained ON-ganglion cells from the mudpuppy retina were studied with a combined approach, including intracellular and extracellular recording from the superfused retina-eyecup preparation, pharmacology with bath-applied 2-amino-4-phosphonobutyrate (APB), and retrograde and intracellular staining using horseradish peroxidase (HRP). Bath application of micromolar levels of APB selectively blocks the light response of ON-bipolar cells; APB was used to separate synaptic inputs into those which originate from ON- vs. OFF-bipolar cells. This approach clearly demonstrates that the light response of the vast majority of sustained ON-ganglion cells is primarily the result of sustained excitatory inputs that arise (directly or indirectly) from ON-bipolar cells. APB revealed the presence of transient excitatory OFF-inputs in many sustained ON-ganglion cells that are normally not evident. Five sustained ON-ganglion cells were intracellularly stained with HRP and their morphology was analyzed with the aid of a computer-assisted neuron reconstruction system. The stained cells are anatomically similar, based on quantitative analysis of a number of morphological parameters. The dendritic trees of all five cells are primarily confined to sublamina b of the inner plexiform layer, although some cells have a small number of processes that ramify in sublamina a. These latter processes may relate to the transient excitatory OFF-inputs revealed with APB application. Ganglion cells which are morphologically similar to the stained, intracellularly sustained ON-ganglion cells were found in a collection of Golgi-like cells that were labeled by retrograde HRP transport. This raises the possibility that sustained ON-ganglion cells in the mudpuppy may constitute a morphologically identifiable class of retinal ganglion cells in this species. There is also some suggestion that a morphologically similar class of OFF-cells may be present.