Amplifying and ameliorating light avoidance in mice with photoreceptor targeting and CGRP sensitization.

Objective: To determine the photoreceptor basis of light avoidance in mice and assess the effect of CGRP sensitization on this behavior. Background: Prior studies have suggested that photophobia is mediated by a subset of retinal ganglion cells (RGCs) that contain melanopsin, making them intrinsically photosensitive (ipRGCs). These cells also receive extrinsic input from cones, which can also mediate light sensitivity. Here, we examined whether spectral variation targeting melanopsin or specific cone types in mice could effectively model light sensitivity. Also, we assessed whether sensitizing mice with calcitonin gene-related peptide (CGRP) could amplify ipRGC-mediated light avoidance. Methods: Light avoidance behavior was observed in a two-zone chamber illuminated by narrow-band LEDs targeting photopic opsins: 365 nm (UV; rodent S-cone), 460 nm (blue; melanopsin), and 630 nm (red; human L-cone). In a non-targeted assay, we assessed the degree of light avoidance in wildtype C57BL/6J mice to varying intensities (5 to 100%) of the blue and red LEDs. In a targeted assay, mice were given a choice to spend time between zones with differing relative contrast levels (0.50, 0.75, or 1.00) for the targeted photoreceptor(s). This was assessed in two transgenic mice with: 1) human red cone knock-in (RCKI), or 2) adult-onset ablation of M1 ipRGCs ( Results: Wildtype mice showed greater light avoidance with increasing light intensity, demonstrating a parametric response. RCKI mice showed avoidance of the high melanopsin (1.00: 0.52 ± 0.08; n = 18) and L-cone (1.00: 0.30 ± 0.11; n = 15) contrast zones but showed a preference for the higher S-cone (1.00: -0.35 ± 0.06; n = 16) contrast zone. These effects decreased with less relative contrast and, thus, contrast dependent. Adding S-cone contrast opposed avoidance to melanopsin (0.10 ± 0.12; n = 14) or L-cone (-0.19 ± 0.10; n = 15) contrast. Ablation of ipRGCs in Conclusions: Our findings further support that ipRGCs play a key role in mediating photophobia. This aversive response to light stems from ipRGCs combining excitatory input from intrinsic melanopsin stimulation and extrinsic L-cone input, which can be opposed by extrinsic inhibitory S-cone input. Chronic exposure to CGRP is likely one of many mechanisms in migraine that can amplify ipRGC signals, leading to photophobia. Plain Language Summary: To better understand light sensitivity, we studied which cells in the eye cause mice to avoid light. We found that mice avoided blue and red light but preferred UV light, and this is the result of a special cell (ipRGCs) in the eye that combines these light signals. Repeated exposure to CGRP, a key nervous system messenger in migraine, increased avoidance of blue light, which may model what happens in people with chronic migraine who experience light sensitivity.