The influence of HEPES on light responses of rabbit horizontal cells

HEPES-buffered solutions, mostly used in studies of isolated cells, and bicarbonate-buffered solutions, mostly used in studies of isolated retinal tissues, have both been used to superfuse an isolated rabbit retina preparation. The responses of horizontal cells (HCs) to light, detected by intracellular microelectrodes filled with Lucifer Yellow, were recorded. Buffering of the superfusate with 100% HEPES completely, but reversibly, abolished the responses of A-type HCs, and is not, therefore, suitable for studies on isolated rabbit retinas. The responses remained when buffering was partially with HEPES and partially with bicarbonate, but were changed: in A-type HCs the overshoot was reduced and the afterpotential was increased. The overshoot may be caused by feedback of HCs on the cones and might be dependent on pHi at the synaptic structure between HCs and photoreceptors.