Extended Data Fig. 2: Identification of candidate ArcAgRP targets and optogenetic activation of projections.
From: Integration of hunger and hormonal state gates infant-directed aggression
a, Immunostainings against c-Fos in brain sections from Agg+ and Agg– mice, and registration to Allen Brain Atlas (see Methods). b, Example FOS+ cell densities in MPOA of Agg+ and Agg− mice in MPOA sections. Scale bars, 200 µm. c, Density of FOS+ cells in hypothalamic brain areas of Agg+ and Agg– mice (n = 6 (Agg+), 5 (Agg–) mice). d, ArcAgRP projections. Areas with significantly different FOS+ cell numbers between Agg+ and Agg− groups, and receiving direct ArcAgRP projections, are highlighted. Note that ADP, MPN, MPO, PD and PS are MPOA subregions. e, Implantation sites of optical fibres in MPOA, LPOA and PVH of Agrp-Cre mice injected with AAV-DIO-ChR2-EYFP. Scale bars, 200 µm. f, Optogenetic stimulation paradigm to assess food intake. g, Correlation between food intake and pup attack latency in mice with optogenetic stimulation of ArcAgRP→MPOA projections (linear regression, R2 = 0.784; P = 0.046, n = 5 mice). h, Latency of food intake during continuous stimulation of ArcAgRP→MPOA and ArcAgRP→PVH projections (n = 4 (MPOA), 4 (PVH) mice). i,j, Effect of optogenetically activating ArcAgRP→MPOA projections (i, 1-h pre-stim, n = 5 mice) on 1-h food consumption in sated mice, and negative controls (j, Agrp-Cre mice injected with AAV-DIO-EYFP, n = 6 mice). k,l, Effect of optogenetically activating ArcAgRP→LPOA (k, n = 6 mice) or ArcAgRP→PVH (l, n = 5 mice) projections on 1-h food consumption in sated mice. m, Pup attack latency after chemogenetic activation of ArcAgRP neurons (Gq), optogenetic stimulation of ArcAgRP→MPOA projections (Opto, 30-min pre stimulation), and food deprivation (n (left to right) = 7, 5 and 39 mice). Statistics: U test in c,h (two-sided, Benjamini-Hochberg adjustment in c). Repeated measures ANOVA in i–m, with pairwise t-tests and Benjamini-Hochberg adjustment in i,j,l. Data are mean ± s.e.m. Box plots: median (line), interquartile range (box), whiskers, 1.5× IQR. See Supplementary Table 1 for acronyms of brain areas. *P < 0.05, **P < 0.01, ***P < 0.001. See Supplementary Table 3 for further details of the statistical analyses.
