Supplementary Figure 13: CRH microinjections into the VTA decrease anxiety at specific doses. | Nature Neuroscience

Supplementary Figure 13: CRH microinjections into the VTA decrease anxiety at specific doses.

From: Chronic CRH depletion from GABAergic, long-range projection neurons in the extended amygdala reduces dopamine release and increases anxiety

Supplementary Figure 13

Bilateral injections of CRH into the VTA of wildtype C57BL/6J mice at (a) 0.4, (b) 40 and (c) 400 ng/side. No behavioral alterations were observed at the lowest CRH dose (unpaired two-tailed t-test / OF: distance t(16) = 0.71, p = 0.49; inner zone time (%) t(16) = 0.18, p = 0.86; n = 8 Vehicle, 10 CRH / DaLi: lit zone time (%) t(15) = 0.75, p = 0.46; no. lit zone entries t(15) = 0.23, p = 0.82; n = 8 vehicle, 9 CRH / EPM: open arm time (%) t(11) = 0.58, p = 0.57; open arm entries (%) t(11) = 0.14, p = 0.89; n = 7 vehicle, 6 CRH). Microinjections of 40 ng/side induced decreased anxiety-related behavior specifically in the EPM (unpaired two-tailed t-test: t(9) = 3.95, **p = 0.003, n = 5 vehicle, 6 CRH) without alerting locomotion or cued fear conditioning (unpaired two-tailed t-test / OF: distance t(11) = 0.52, p = 0.66; inner zone time (%) t(11) = 0.60, p = 0.56; n = 6 vehicle, 7 CRH / DaLi: lit zone time (%) t(13) = 1.4, p = 0.19; no. lit zone entries t(13) = 0.93, p = 0.37; n = 7 vehicle, 8 CRH / EPM: open arm entries (%) t(11) = 0.66, p = 0.53; n = 5 vehicle, 6 CRH / fear conditioning (RM ANOVA) – tone/auditory: time x genotype interaction F6,72 = 1.83, p = 0.11; genotype F1,12 = 0.42, p = 0.69 / context: time x genotype interaction F2,24 = 0.53, p = 0.59; genotype F1,12 = 0.40, p = 0.75; n = 7 vehicle, 7 CRH). At the highest dose of 400 ng/side, CRH induced a pronounced decrease in locomotion in the OF (unpaired two-tailed t-test / OF: distance - t(15) = 3.5, **p = 0.003; inner zone time (%) t(15) = 1.83, p = 0.09; n = 8 vehicle, 9 CRH) and lead to a decrease in number of lit zone entries in the DaLi (unpaired two-tailed t-test / DaLi: lit zone time (%) t(16) = 0.43, p = 0.67; no. lit zone entries t(16) = 2.5, *p = 0.025; n = 9 vehicle, 9 CRH). (d) Representative coronal brain section (right) and corresponding atlas image (left) depicting the correct placement of the injection cannula in the VTA. Error bars represent s.e.m.

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