Fig. 4

a Schematic depicting the hypothesized telencephalic subpallial (SPa) pathway linking olfactory bulb conspecific cue responses to preoptic OXT neurons. b Top left: Maximum-projection image showing overlap of Tg(y321:Gal4;UAS:Kaede) (green) with GABAergic neurons labeled by Tg(Gad1b:RFP) (magenta) in the telencephalon. y321 and Gad1b co-expression occurs in both the anterior subpallium (aSPa, rostral to the anterior commissure) and posterior commissure (pSPa, caudal to the anterior commissure and rostral to the preoptic area). Top right: Lower-magnification maximum-projection image showing overlap of Tg(y321:Gal4;UAS:Kaede) with Tg(Gad1b:RFP) (magenta). Overall, 88.2 ± 1.5% of aSPa^y321 and 87.3 ± 3.06% of pSPa^y321 neurons are GABAergic (n = 7 fish). OB olfactory bulb, Tel telencephalon, PO preoptic area; PT posterior tuberculum. Bottom: y321 and Gad1b overlap shown across different z-planes. All scale bars = 20 µm. This experiment was repeated on 7 fish with similar results. c OXT neuron dendrites project into the location where SPa^y321 neurons reside. Top: Maximum-projection image; Bottom: 3D projection at different rotation angles. Scale bar = 50 µm. This experiment was repeated on more than 3 fish with similar results. d 2-P calcium imaging was performed on Tg(y321:Gal4; UASGCaMP6s) neurons in the aSPa region on fish exposed to kin, non-kin, and adult water cues. See Supplementary Figs. 4, 5 for imaging and analysis of the same telencephalic regions in Tg(HuC:GCaMP6s; Gad1b:RFP fish). Top left: Maximum-projection image showing the aSPa^y321 region imaged. Scale bar = 50 µm. Top right: Example Δf/f trace of a kin-responsive unit. Bottom: Stimulus-triggered averages for units that are kin-selective, adult-selective, or responsive to multiple conspecific cues. Shaded region indicates SEM. This experiment was repeated on 4 fish with similar results. e K-means clustering (k = 12) was performed on the matrix of correlation coefficients (color coded blue to red) to water and conspecific stimuli n = 4875 units from 4 fish. Number of units within each cluster and percentage representation of total units are displayed on the right. Clusters specific to kin, non-kin, and adult water, as well as clusters with mixed selectivity were observed. f Mean stimulus-triggered Δf/f for each cluster, aligned to stimulus onset. Order of clusters is the same as in (e). Source data are provided as a Source Data file.