Effects of Test Flash Duration on the Photopic Negative Response (PhNR) of the Flash Electroretinogram

Abstract

Purpose: The Photopic Negative Response (PhNR) of the cone mediated electroretinogram (ERG) is a slow potential with negative polarity that appears after the b-wave. The PhNR originates from the electrical activity of retinal ganglion cells (RGCS) and has clinical utility. The PhNR is typically recorded to a brief (<4ms) test flash, we explored the effect of increasing the stimulus duration on the PhNR amplitude of normal subjects in an ongoing attempt to optimize the stimulus conditions for its clinical use. Methods: ERGs were recorded with DTL electrodes from normal subjects (N=10) in the age range 23-53 years using the ColorBurst handheld ganzfeld stimulator and hardware from Diagnosys (Lowell, MA). The stimuli consisted of red test flashes on constant blue background (8 phot cd.s/m.sq). The test flashes were either brief stimuli (<4 ms duration) in the range of 0.00625 - 6.4 phot cd.s/m.sq or longer duration (20-80 ms) in the range of 0.125-1500 cd/m.sq. A new algorithm in the Espion software with objective sweep selection based on various noise and artifact identification criteria was used to average repeated responses at each test flash intensity. The PhNR amplitude of the averaged waveform was plotted as a function of test flash intensity and fitted with the standard Naka-Rushton equation. The saturated amplitude (Vmax), slope (n) and semisaturation constant (K) derived from the fits were analyzed. The student t-test was performed to compare the fit parameters across different test flash durations with correction for multiple comparisons using the Holm’s method. Results: Vmax for the brief stimulus was 20+7 microvolts and increased to 23+2 microvolts for 20 ms duration stimuli. With further increase in stimulus duration the PhNR Vmax was 34+8 v, 42+10 v and 37+10 v for 40 ms, 50 ms and 60ms duration stimuli and thereafter reduced to 29+7 v for an 80 ms stimulus duration. The Vmax amplitude differences between the brief and longer duration stimuli were statistically significant only for the 40 ms (p=0.04), 50 ms (p=0.001) and 60 ms (p=0.01) after correcting for multiple comparisons. Responses to stimulus durations up to 60 ms demonstrated a single PhNR trough and for longer duration stimuli two PhNR troughs were observed one following light onset after the b-wave and another following light offset. Conclusions: The saturated PhNR amplitude is larger for longer duration stimuli and is maximal in the range of 40-80 ms duration. The larger PhNR amplitude at the intermediate test flashes likely reflect the summation of the PhNR to stimulus onset and offset and could potentially have more value in assessing retinal ganglion cell function in patients with disease affecting the optic nerve and/or inner-retinal neurons.