Silent-substitution stimuli silence the light responses of cones but not their output

Chromatic vision starts at the retinal photoreceptors but photoreceptors are themselves color-blind, responding only to their effective quantal catch and not to the wavelength of the caught photon per se. Mitchell and Rushton (1971) termed this phenomenon the univariance concept, and it is widely used in designing silent-substitution stimuli to test the unique contributions of specific photoreceptor types to vision. In principle, this procedure controls the effective quantal catch of photoreceptors well and hence works at the phototransduction-cascade level of vision. However, both phototransduction-cascade modulation and the horizontal-cell-mediated feedback signal determine photoreceptor output. Horizontal cells receive input from, and send feedback to, more than one photoreceptor type. This should mean that silent-substitution stimuli do not silence horizontal-cell activity, and that this activity is fed back to the silenced cones. This in turn will modulate the output of silenced cones, making them not so silent after all. Here we tested this idea and found that silent-substitution stimuli can adequately silence cone-membrane potential responses. However, these cones still received a feedback signal from horizontal cells, which modulates their Ca2+ current and thus their output. These feedback-induced Ca2+-current changes are substantial, as they are of the same order of magnitude as Ca2+-current changes that occur when cones are directly stimulated with light. This illustrates that great care needs to be taken in interpreting results obtained with silent-substitution stimuli. In the discussion, we outline two basic types of interpretation pitfalls that can occur.