a, Schematic of stimulus presentation for activity mapping. b, Attraction towards stimuli shown in a. n = 17 (no stimulus) or n = 9 (continuous; bout-like) single animals; and n = 8 animals tested in 4 pairs (conspecific). Data are mean (black dots) ± 1 s.d. Exact P values were calculated using two-tailed t-tests compared with the no-stimulus group: P = 0.16 (continuous); P = 5.2 × 10⁻⁸ (bout-like); P = 8.1 × 10⁻¹¹ (conspecific). Bonferroni-corrected α values: NS, P > 0.05/3 (NS); ***P < 0.001/3. c, Representative slices of maximum-intensity-normalized c-fos signal merged across all 28 registered animals. The views are horizontal (top row), sagittal (bottom left) and coronal (bottom right). The solid grey lines indicate the corresponding planes across the slices. The dashed line indicates the midline. Coloured patches indicate activity clusters (Extended Data Fig. 1). A, anterior; D, dorsal; L, lateral. d, The average normalized c-fos signal at the three representative horizontal planes indicated in c. n = 6 (no stimulus), n = 8 (continuous), n = 6 (bout-like) and n = 8 (conspecific) animals. e, The effect size (Cohen’s d) of normalized bulk c-fos induction compared with the no-stimulus condition. Negative values indicate a lower signal compared with the no-stimulus condition. The dendrogram represents hierarchical clustering. Statistical analysis was performed using two-tailed t-tests in each activity cluster versus the no-stimulus group. *P < 0.05/3, **P < 0.01/3, ***P < 0.001/3 (α values were Bonferroni-corrected per activity cluster). Animal numbers are the same as in d. Additional statistical information is provided as Source Data. Scale bars, 200 µm. DT, dorsal thalamus; En, entopeduncular nucleus; Hc1, caudal hypothalamus 1; Hc2, caudal hypothalamus 2; Hc3, caudal hypothalamus 3; Hi1, intermediate hypothalamus 1; Hi2, intermediate hypothalamus 2; Hi3, intermediate hypothalamus 3; Hrl, rostral hypothalamus, lateral; mHr, rostral hypothalamus, medial; MOd, medulla oblongata, dorsal; MOi, medulla oblongata, intermediate; nMLF, nucleus of the medial longitudinal fasciculus; OB, olfactory bulb; P, pallium; Pl, pallium, lateral; PM, magnocellular preoptic nucleus; Pn, pineal; PPa, anterior parvocellular preoptic nucleus; PPp, posterior parvocellular preoptic nucleus; Pr, pretectum; PT, posterior tuberculum; Ri, inferior raphe; SPd, subpallium, dorsal; SPv, subpallium, ventral; TeOa, tectum, anterior; TeOd, tectum, dorsal; TeOv, tectum, ventral; Tg, lateral tegmentum; TS, torus semicircularis; VT, ventral thalamus.

Source data

a, Schematic of the experimental set-up. b, Example imaging planes in the TeO and DT with all segmented neuronal ROIs, representative for n = 11 animals (left). Right, representative normalized ∆F/F traces of one tectal and one thalamic neuron. c, Horizontal view of all responsive neurons (n = 28,306 total, 2,573 ± 1,175 per fish) from 11 fish (18–22 d.p.f.) aligned to a juvenile reference brain (left). Colour indicates mean baseline ∆F/F (no stimulus), and responses to continuous and bout-like motion. The yellow line indicates the DT. Right, mean responses of all DT neurons per fish (n = 258 ± 198, 2,837 total) from n = 6 animals with a number of recorded DT-BPNs of >30. Data are mean ± 1 s.d. d, Mean ∆F/F responses of example neurons from b to all stimulus frequencies. e, The distribution of all responsive neurons from n = 11 fish in the reference brain. The colour map shows the BPI. Opacity scales with absolute BPI (0–0.5). f, The distribution of BPNs (312 ± 143 neurons per fish, 3,437 total). Colour reflects a Gaussian KDE; contours delineate densities of 0.1, 0.15 and 0.3 BPNs per 1,000 μm³. n = 11 fish. g, DT-BPN tuning to stimulus frequency. The mean peak across neurons was 1.2 Hz  ± 1.6 Hz. n = 563 neurons. The black lines represent the mean values of individual animals. Data are from a subset of animals in e with a number of recorded DT-BPNs of >30. n = 6 animals. h, DT-BPN tuning to average speed at 1.5 Hz or 60 Hz and acceleration. The cartoons show stimulus displacement over time. Data are mean ± 1 s.d. of all of the neurons shown above. n = 291 neurons. The black lines indicate individual animals. n = 4 fish, 73 ± 10 neurons per fish. i, DT-BPN and PreT responses to local dot motion and whole-field motion and their anatomical distribution. Circles (left) show the mean of individual animals. n = 4 fish, 77 ± 15 (DT-BPNs), 114 ± 48 (PreT) neurons per fish. Data are mean ± 1 s.d. j, The distribution of BPNs in 7 d.p.f. larvae (n = 4 fish, 230 ± 87 neurons per fish) as in f. Scale bars, 100 μm (b and i) and 200 μm (c, e, f and j).

Source data

a, Frontal view of an EM reconstruction of neurons in the bout-preference region (BPN KDE, red) of the DT. Axons are shown in blue. b, Top view of the neurons shown in a. c, Magnified view of the thalamic arborization field, outlined in b. Synapses with identified presynaptic partners are shown as blue spheres. One representative synapse of a tectal PVPN axon (white arrow) onto a putative BPN’s dendrite (pBPN, black arrow) is indicated below (randomly chosen). d, Frontal view of tectal PVPNs (green) and their postsynaptic pBPN partners (red). e, Example of a single PVPN (green), which makes ipsilateral synaptic contacts to at least four identified pBPNs (red). f, Side view of the left (top) and the right (bottom) tectal SFGS layers, showing the PVPNs (green) and their presynaptic retinal ganglion cell axons (different colours). PVPN axons are not shown for clarity. g, Side view of the pBPNs (red, axons in blue) and their axonal target regions (Supplementary Video 2). AF4 (yellow) is shown as a reference. h, Circuit diagram. Identified cell types are indicated next to arrows with cell numbers in parentheses. For c, scale bar, 0.5 µm.

a, Schematic of the shoaling test after chemogenetic ablation. ctr., control. b, Two-photon image of 21 d.p.f. SAGFF(lf)81c:Gal4, UAS:NTR-mCherry and elavl3:H2B-GCaMP6s animals 24 h after ablation versus control treatment. Representative of three similar fish. c,d, Reduced neighbour density (c) and attraction (d) in 81c:NTR-ablated animals. Short-range repulsion is intact. n = 13 (ablated) and n = 15 (control) animals. The neighbour maps in c show the probability of finding the stimulus in space with the animal at the centre of the map, heading up. Each map is 60 mm × 60 mm. AU, arbitrary units. e, Schematic of volumetric two-photon imaging in the DT after 81c:NTR ablation. f, Mean and example ΔF traces of all DT-BPNs for ablated and control animals show that 81c:NTR ablation strongly reduces responses to bout-like motion. The vertical lines mark the start of stimulus presentation. The shaded areas denote 1 s.d. around the mean. n = 25 (ablated) and n = 849 (control). g,h, Fewer DT-BPNs in 81c:NTR (-ablated) animals. g, The anatomical location of all DT-BPNs across animals coloured by mean ΔF/F to bout-like motion. n = 25 (ablated) and n = 849 (control). The yellow area shows the DT. h, Quantification of DT-BPNs per animal. n = 7 (ablated) and 9 (control) animals. Cohen’s d effect size is shown. The P value was calculated using the two-sided Mann–Whitney U-test. i, Attraction is strongly reduced in s1026tEt:NTR-ablated animals. Short-range repulsion is intact. n = 7 (ablated), n = 21 (control) animals. j, Bilateral two-photon laser ablation of neurons in the DT-BPN region in juvenile zebrafish. k, Reduced attraction after DT laser ablation. n = 7 (ablated) and n = 9 (embedding control) animals. Short-range repulsion is intact. Data in d, h, i and k represent individual animals and mean ± 1 s.d. Cohen’s d effect size is shown. The P values were calculated using two-tailed Student’s t-tests with no correction. For b, e, g and j, scale bars, 100 µm.

Source data.