The objective of the present work was to determine the interaction of
cone inputs in the response of horizontal cells using heterochromatic
flicker photometry (HFP). Intracellular electrophysiological recordings
were made in horizontal cells of isolated retinae of carp maintained in
physiological solution, with the receptor side up. Sharp glass
microelectrodes filled with 3 M KCl solution with resistances between 100
and 120 MΩ were used. Stimuli comprised six cycles of two 6-Hz
sinusoidal light waves in counterphase adjusted for the same number of
quanta: a green light (550 nm) from a monochromator with a Xenon lamp and
an LED red light (628 nm). The stimulation program consisted of 10 steps
with the 550-nm wave at constant amplitude, while the 628-nm wave varied
in increments of 10% up to 100%, followed by another 10 steps with the
628-nm wave at constant amplitude while the 550-nm wave varied in
increments of 10% up to 100%. We recorded responses from four different
horizontal cell classes: H1 (monophasic, broadband, n = 37), H2
(biphasic, red-green color-opponent, n = 13), and H3 (biphasic,
blue-yellow color-opponent, n = 2) cone horizontal cells; and RH
(monophasic, broadband, n = 3) rod horizontal cells. H1 and RH
horizontal cells showed a similar cancellation point at a heterochromatic
mixture consistent with mixed inputs from 630- and 550-nm cones. No
cancellation point was found for the H2 cell class. Fish H1 cells add cone
inputs and signal “luminance” in light levels appropriate for
cone stimulation. The same occurs with RH cells, which also signal
“luminance,” but in light levels appropriate for rod work. For
both cell classes there is an HFP cancellation point occurring at a
combination of 628-nm and 550-nm lights in opposing phase that leads to
the cancellation of the cell's response. No cancellation was found
for H2 and H3 cells, which are the chromatically opponent horizontal cells
in lower vertebrates.