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. 2021 Aug 19;184(17):4564–4578.e18. doi: 10.1016/j.cell.2021.07.002

Figure S7.

Figure S7

Optogenetic perturbation of SN-projecting MLR neurons, related to Figure 6

(A) Effect of laser stimulation on normalized rolling s.d. of position (mean ± SEM) of body parts (forelimbs, hindlimbs, snout) upon laser application in MLR-Rbp4 loss-of-function (LOF) (n = 10; left) and control mice (n = 5, right) and bar plot comparing light-induced change in the normalized s.d. of position (mean ± SEM) in MLR-Rbp4 LOF (n = 10) and control mice (n = 5).

(B) Anatomical confirmation of fiber placements for MLR-Rbp4 LOF mice at corresponding rostro-caudal levels (Bregma indicated).

(C) Electromyography in the forelimb muscles biceps and triceps during ongoing locomotion and upon optogenetic stimulation of MLR-Rbp4 neurons in an example mouse (blue window: laser application; red line: speed trace). Note that behavioral arrest is accompanied by cessation of biceps and triceps alternating muscle contraction.

(D) Comparison of the change in speed (mean ± SEM) between the MLR-Rbp4 gain-of-function (GOF; n = 10) and control mice (n = 5) upon light application.

(E) Effect of light application on normalized rolling s.d. of position (mean ± SEM) for different body parts (as indicated: forelimbs, hindlimbs, genitals, tail base, snout) for rearing (left), grooming (middle) and handling (right) trials upon laser application in MLR-Rbp4 GOF (n = 10) and control mice (n = 5). The bar plots indicate the comparison of the light induced change in the normalized rolling s.d. of position (mean ± SEM) for each behavior between MLR-Rbp4 GOF (n = 10) and control mice (n = 5).

(F) Top: Experimental strategy for optogenetic activation of MLR-Rbp4 neurons (n = 10), axonal terminals of vGlut2-MLR neurons over SN (n = 10), vGlut2-MLR neurons with projections to SN (n = 7), compared to control mice (n = 8) not expressing optogenetic activators. Middle: Graphs from example mice showing speed versus time plots and depict single trials (in gray) and their averages for closed loop laser application (red), no laser application (black), as well as an overlay of these two experimental conditions. Bottom: Anatomical confirmation of fiber placements for analyzed mice at corresponding rostro-caudal levels (Bregma indicated; one fiber of one animal not shown due to unilateral dorsal fiber placement).

∗∗p ≤ 0.01.