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. Author manuscript; available in PMC: 2014 Nov 29.
Published in final edited form as: Nature. 2014 Apr 16;509(7502):627–632. doi: 10.1038/nature13169

Figure 2. Esr1+ cells in VMHvl are necessary and sufficient for aggression.

Figure 2

a. Strategy for optogenetic activation of Esr1+ cells in VMHvl. EF1α, elongation factor 1α promoter; ChR2 is V5 epitope-tagged. b-d. Whole-cell patch clamp recording from Esr1+ cells in VMHvl (c, EYFP+ cell) in acute hypothalamic slices. Photostimulation-evoked spiking (d, upper) and quantification of spike fidelity (d, lower) are shown (filled circles, 2 ms light pulse-width, n=11 cells; open circles, 20 ms pulse-width, n=5 cells). e-j. Double-labeling for Cre-dependent hrGFP viral reporter (e, h) and c-Fos (f, i) in VMHvl of Esr1cre/+ males following photostimulation; h-j, boxed areas indicated in e-g. k. Quantification of (e-j) (mCherry, n=5; ChR2, n=10; ***p<0.001; Mann-Whitney U-test). l, m. Representative raster plots (l) and video stills (m) illustrating photostimulation-evoked (blue bars; mW/mm2) attack (l, red rasters) or close investigation (CI, yellow rasters) byChR2-expressing Esr1cre/+ males (m, black mice), toward a castrated male (”♂”; l, m, upper) or an intact female (l, m, lower). See Supp. Video 1. n-q. Quantification of attack parameters towards castrated males (ChR2, n, n=33; o, n=23; p-q, n=11; mCherry, n, n=14) or females (ChR2, n, n=28; o. n=22; p-q. n=16; mCherry, n, n=10). r. Photoactivation of Esr1- cells using “Cre-out” ChR2 rAAV. s. Percentage of wild-type (WT; n=4) and Esr1cre/+(n=9) males showing photostimulated attack toward castrated males (see ED Fig. 3b, c). t. Expression of eNpHR3.0 in VMHvl Esr1+ neurons. u. Whole-cell patch clamp recording in acute hypothalamic slices, showing photostimulation-induced suppression of current injection-evoked spiking in eNpHR3.0-mCherry expressing Esr1+ cells. v. Representative raster plots illustrating effect of photostimulation on attack towards a male intruder. w-y. Quantification of behavioral parameters (mCherry, n=10; eNpHR3.0, n=13; ****p<0.0001; Mann-Whitney U-test). z. Percentage of photostimulation trials in which approach to intruder led to attack (mCherry, n=7; eNpHR3, n=4; **p<0.01; Mann-Whitney U-test). Data are mean ± SEM (d, k, o, z) or median ± min and max values (p-q, w-y).