Fig. 3

Fig. 3

a Left: Schematic of imaging and cue-delivery setup. Two-photon (2-P) calcium imaging of the olfactory bulb was performed as each Tg(HuC:GCaMP6s) fish was presented with pulses of kin water, non-kin water, and adult water (randomized order), alternated with pulses of water. Right: Stimulus-triggered average of all olfactory bulb (OB) neuron responses (Δf/f) to kin, non-kin, and adult water. b Δf/f traces (bottom, raster plot for an example fish) from individual units (see Methods) within the OB were correlated with regressors (top) fit to each stimulus type. c Mean OB Δf/f integrated over a 60-s period post stimulus from a single fish (same fish shown in Fig. 3b, also see Supplementary Fig. 3), shown for each plane (ventral to dorsal) and each cue in a volumetric stack. Bottom-most panel shows the anatomy references for each plane. A anterior, L left. Scale bar = 50 µm. This experiment was repeated on 10 fish with similar results. d Δf/f traces from individual units were extracted from the OB (see Methods). Top: Correlation coefficients for each unit (n = 477 units from the same fish in b and c) with each regressor are color coded (blue to red) and overlaid over the anatomy image per z-plane. Bottom: Each unit’s response (i.e., correlation coefficient) to water and conspecific stimuli, from the same fish as above, clustered using K-means clustering (as in e). 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 = 4018 units from 10 fish). Number of units within each cluster and percentage representation of total units are displayed on the right. Clusters specific to kin water 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). As OB responses are often sustained across the entire stimulus epoch, negative Δf/f and correlation coefficients in response to water flow are likely due to washout of residual olfactants, rather than an active suppression of the signals by water flow. g 3D plot displaying the spatial localization of units within each cluster, for a few selected clusters (all clusters shown in Supplementary Fig. 3c). XY coordinates for all units per fish were scaled linearly to their minimum and maximum values in each dimension. Adult-responsive clusters tend to be situated laterally (e.g., cluster 7), whereas clusters that show more general conspecific responses (e.g., clusters 4,5,10) are situated more medially. Kin-specific responses (clusters 1 and 2) are more scattered, though they extend more laterally than general conspecific-responsive clusters. A anterior, P posterior, D dorsal, V ventral. Source data are provided as a Source Data file.