Figure 1.

IPL synaptic density/volume is decreased 12 dpi

(A) Representation of an EAE score graph in a 60-day experiment (arrows indicate the timing of tissue collection). Synaptic density was calculated by analyzing the area in proximity of the optic nerve head for 12 EAE female mice and 9 sham-immunized mice at onset (12 dpi, four EAE mice and three sham-immunized mice), peak of disease (18 dpi, four EAE mice and three sham-immunized mice), and at a chronic stage (60 dpi, four EAE mice and three sham-immunized mice).

(B–I) Staining for presynaptic inputs (Bassoon and VGluT1, green) and postsynaptic compartments (Homer 1, red) and colocalization of pre-and postsynaptic terminals (yellow) in sham-immunized mice (B and F), at 12 dpi (C and G), 18 dpi (D and H), and 60 dpi (E and I). Scale bar: 10 um.

(J–K) Synaptic counts per area showed a trend toward reduction on day 12 and day 18 and on day 60 showed statistically significant synaptic loss.

(L–O) Examples of retinal IPL segmentation from DAPI images: sham-immunized mice (L), 12 dpi (M), 18 dpi (N), and 60 dpi (O). Scale bar: 100 um.

(P and Q) Quantification of the number of functional synapses per IPL volume, given by colocalization of both Bassoon/Homer (P) and VGlut1/Homer (Q) per IPL volume. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Technical replicates: for each animal, 3–4 stained retinal cross-sections at the level of the optic disc containing the central retina.