In the vertebrate retina, visual information is segregated into an on channel excited by light increment and a complementary off channel excited by light decrement. We used 2-amino-4-phosphonobutyric acid (APB), which selectively blocks the on channel in the retina (29), to determine the contributions of the on and off pathways to response properties of neurons in the lateral geniculate nucleus (LGN) of anesthetized, paralyzed rabbits. Visually evoked responses were recorded from 46 single cells in the LGN before, during, and after vitreal perfusion with 200-1,000 microM APB. APB reversibly blocked responses of on uniform-field cells and on center concentric-field cells to stationary, flashing spots of light. Responses to off uniform-field cells and off-center concentric-field cells were largely unaffected. APB did not differentially affect responses elicited from the receptive-field centers, as opposed to the surrounds, of on-center concentric-field cells. This finding suggests that these cells are driven exclusively by the on retinal channel and that the center-surround organization of their receptive fields does not result from a convergence of the on and off pathways. We studied a small number of cells that were selective for stimulus direction or motion. In each case, APB eliminated the cell's response to a moving light edge. The surviving response to a moving dark edge retained its original direction or motion preference, suggesting that these response properties do not depend critically on interactions between the on and the off pathways. The findings obtained in the rabbit are reminiscent of the results of similar investigations in the cat (10, 11) and the monkey (25). Taken together, they indicate that in the LGN of several vertebrate species there is a precise segregation vertebrate species there is a precise segregation of on and off information, at least for some functional classes of cells. The combination of on and off information does not seem to play a major role in establishing the response properties observed at this level in the visual system.