FullFull -Field Ch Chro roma matic tic Pupillometry in the Assessment of the PostPost-Illumination Pupil Response Driven by MelanopsinMelanopsin-Containing

Retinal Ganglion Cells No.4110 - A0098

Session: 413

Lei, 1,2H.C. Goltz, 1M. Chandrakumar, 1,2,3A.M.F. Wong 1Program in Neuroscience & Mental Health, Hospital for Sick Children; Department of Ophthalmology and Vision Sciences, 2University of Toronto & 3Hospital for Sick Children, Toronto Canada

1Shaobo

Introduction
Melanopsin is a photopigment allowing a small subset of
retinal ganglion cells to absorb light energy directly and fire action potential with or without synaptic input form rods and cones. Intrinsic photo response of melanopsin-containing retinal ganglion cells (MGCs) provides sustained irradiance coding for non-visual photoperception functions including circadian photoentrainment and the pupillary light response. A sustained pupil constriction can be observed after the offset of a bright blue light stimulus. This post-illumination pupil response (PIPR) is produced by MGC, and can be measured by chromatic pupillometry. Measuring PIPR holds promise as a novel tool to assess MGC function. Current testing protocols use central-field stimulation and require a very bright light of long duration which can be difficult for some subjects. We test the hypothesis that a consistent PIPR can be induced with full-field blue light stimuli of shorter duration and lower intensity than with central-field stimulation.

Experimental Conditions (cont.):

For each of the intensity and duration steps, a red flash was presented first followed by a blue flash 45 s after the offset of the red flash. Participants were provided with a short break at 45 s after the offset of the blue flash to allow the pupil size to return to baseline and to prevent fatigue.

PIPR to full-field blue stimulation increased monotonically with increasing stimulus intensity. Red light (internal reference) induced no or minimal PIPR at all intensity level. Full-Field vs. Central-field Stimulation

Measurement and Data Analysis:

Pupil size data were normalized to baseline calculated from the mean pupil size during a 1 s period before the onset of each stimulus. Primary Outcome Measure PIPR: mean of the normalized pupil size over a 20 s period from 10-30s after the offset of light stimuli (smaller number means greater pupil constriction).
* comparison of PIPR to full-field blue stimuli of 400 cd/m2 intensity of the 6 longest duration steps (100, 200, 400, 600, 800, and 1000 ms. * Duration-response function of PIPR to full-field stimulation vs. PIPR to 1s, 400 cd/m2 central-field stimuli

No further increase in PIPR was observed when the duration increased from 400-1000 ms. 400-1000 ms, 400 cd/m2 blue full-field stimuli induced significantly greater PIPR than 1000 ms blue central-field stimulus of the same duration.

* Comparison of PIPR induced using 400 cd/m2 central-field stimuli (dashed line) vs 100-400 cd/m2 full-field stimuli (solid lines)

* Intensity-response function of PIPR to full-field stimulation vs. PIPR to 1s, 400 cd/m2 central-field stimuli

Methods
Apparatus:
Full-field and central-field light stimulation were presented with a Colordome Ganzfeld bowl originally designed for ERG testing. Pupillary light response was monitored with a video based eye tracker at 60Hz. Statistic Analysis One-way ANOVA with post hoc corrected for pairwise multiple comparisons using the TukeyKramer method.

400 cd/m2, full-field blue stimulus induced significantly larger PIPR than central-field blue stimulus of the same intensity. PIPR to red flashes did not differ significantly full-field vs. central-field.

Conclusions
Full-field stimulation is more effective that central-field stimulation in inducing PIPR, suggesting that PIPR is a function of stimulus intensity, duration and area. This study is the first to demonstrate that saturating PIPR up to 30 seconds post illumination can be induced in vivo with a strong blue flash lasting only a few hundred milliseconds. This updated understanding of the relation between PIPR and stimulus intensity, duration and area will allow investigators to tailor their PIPR testing paradigm to target a specific research question, and greatly facilitate the development of a convenient and comfortable technique to assess MGCs function for emerging clinical use.

Participants:
10 visually-normal adult subjects (mean age: 31 years, range 22 56), only right eyes were stimulated and recorded, left eyes were patched during recording.

Results: Experiment 2 (duration trials)

Results: Experiment 1 (intensity trials)

Experimental Conditions:
Experiment 1 (intensity trials): After 10 min dark adaptation, PIPR was induced with alternating red (64010 nm) and blue (46717 nm), 1-second full-field stimuli of increasing intensity from 0.1 to 400 cd/m2 (11 steps) . For comparison with a previously published protocol, a 6090 central-field blue stimulus at 400 cd/m2 was also presented for 1 second. Experiment 2 (duration trials): 10 min dark adaptation, 100 cd/m2 and 400 cd/m2, red and blue full-field stimulations of increasing duration from 4-1000 ms were presented alternately. 100 cd/m2 and 400 cd/m2 trials were conducted on different days.
* Mean (n=10) normalized postillumination pupil response (PIPR) tracings in response to 1 s stimulation of varying intensity from 10 visually-normal participants.

* Mean PIPR to 100 cd/m2 and 400 cd/m2 full-field stimulation of varying duration from 10 visually-normal observers.

Contact Information
shao-bo.lei@sickkids.ca
PIPR in response to 400 cd/m2 increased as the duration of stimulus increased from 4-200 ms. *Authors have no commercial interest to disclose.