The refractive state of hatchling chicks rapidly compensates to applied optical defocus through alteration in eye growth. The mechanism is capable of sensing whether the plane of focus lies in front of or behind the photoreceptors, however, its nature and site of action within the retina are unknown. We attempted to create an imbalance in the adaptation of the retinal ON and OFF mechanisms previously implicated in refractive control through pharmacological interventions, by rearing chicks from 4 to 9 days of age with a monocular +10 D, 0 D or -10 D lens, in an environment illuminated by a moving or stationary plaid of luminance gradients. When the plaid moved in one direction a local Fast-ON sawtooth luminance modulation was produced, while plaid motion in the other direction resulted in a Fast-OFF sawtooth modulation. Significantly reduced refractive compensation accompanied +10 D lens/Fast-OFF and -10 D lens/Fast-ON rearing, but not for the other conditions. Thus the refractive compensation mechanism depends on the nature of the temporal contrast of the environment, suggesting a relationship between the sign of defocus and the state of adaptation of the retinal ON and OFF subsystems.