Fig. 4: Optogenetic inhibition of LHb-DRN projection neurons and instigation-heightened aggression. | Nature Communications

Fig. 4: Optogenetic inhibition of LHb-DRN projection neurons and instigation-heightened aggression.

From: Lateral habenula glutamatergic neurons projecting to the dorsal raphe nucleus promote aggressive arousal in mice

Fig. 4: Optogenetic inhibition of LHb-DRN projection neurons and instigation-heightened aggression.The alternative text for this image may have been generated using AI.

a Schematics and timeline of this experiment. AAV2-hSyn-eNpHR3.0-EYFP was injected into bilateral LHb. Aggression tests were conducted every other day, and RI test and Inst test was conducted alternately. Yellow light (593 nm, 3 mW) was delivered to the DRN on days 7 and 15. b, c Expression of eNpHR3.0-EYFP (green) was observed in bilateral LHb (b scale bar 500 μm). Blue: DAPI. Projection terminals were observed in the DRN (outlined in white lines) and median raphe nucleus (c scale bar 500 μm). d Duration of total aggressive behaviors in each session. Gray lines show each individual’s data (n = 9 biologically independent animals) and black line indicates average data (Mean ± SEM). e Summarized data for the duration of total aggressive behavior in each condition. The Inst test increased total aggressive behavior compared to the RI test in OFF sessions, but light stimulation (ON) blocked the effect of social instigation (one-way repeated measures ANOVA, n = 9 biologically independent animals, F(2,16) = 12.65, p = 0.0005, post hoc t test with Tukey’s multiple comparisons tests (two-sided)). f–i Detailed behavioral analysis showed that optogenetic inhibition of LHb-DRN blocked the pro-aggressive effect of social instigation (one-way repeated measures ANOVA, n = 9 biologically independent animals, attack bites: F(2,16) = 15.47, p = 0.0002 (f), sideways threats: F(2,16) = 18.15, p < 0.0001 (g), post hoc t test with Tukey’s multiple comparisons tests (two-sided)), but had no effect on non-aggressive behaviors such as locomotion (h) and social contact (i) (one-way repeated measures ANOVA, n = 9 biologically independent animals, both n.s.). No effect of light stimulation was observed in the RI test on the duration of total aggressive behavior (j), frequency of attack bites (k), duration of sideways threats (l), locomotion (m), and social contacts (n) (paired t test (two-sided), n = 9 biologically independent animals, all n.s.). Each bar represents mean value ± SEM, and gray line indicates each individual’s data. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Source data are provided as a Source Data file.

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