Extended Data Fig. 10: Effects of impaired sensory input on POA development.

(a) Distance between mutant and control sample centroids in PCA space (normalized to intra-genotype distance) for inhibitory clusters. Control samples included both C57BL6/J and control littermates. Sample numbers, left to right columns, mutant/control: 2/8, 4/10, 2/10, 2/8, 2/8, 2/6, 2/7, 2/7, 2/8, 2/8, 2/7. (b) Random forest classifier approach (Augur) to identify cell types that show differences between mutant and control samples. Each point represents a single cell type. Number of biological replicates as described in (a). (c) Mutant-control PCA distance for all clusters in Trpc2 P10 experiments, split according to sub-region, and ordered from most to least affected. Data from n = 4 biological replicates per genotype. (d) UMAP showing identification of visual cortex neuronal subtypes in Trpc2 mutants and controls (pooled) at P10. (e) Random forest classifier (Augur) for replication of Trpc2 P10 POA (left) and visual cortex (right) from the same brains. Cells pooled from n = 2 biological replicates per genotype. (f) Weight of Trpc2−/− pups and Trpc2−/+ littermates at P7 to P14. n = 41 (Trpc2−/+) and 30 (Trpc2−/−). Error bars show standard error of the mean. (g) Latency for mothers to retrieve Trpc2−/+ or Trpc2−/− P9-10 pups when given a choice between the two. n = 48 (Trpc2−/+) and 48 (Trpc2−/−). (h) Latency for Trpc2−/+ or Trpc2−/− P13-14 pups to first contact their anesthetized mother. n = 26 (Trpc2−/+) and 22 (Trpc2−/−). (i) Latency for Trpc2−/+ or Trpc2−/− P11-12 pups to first contact with littermates. n = 36 (Trpc2−/+) and 20 (Trpc2−/−). (j) Latency for Trpc2−/+ or Trpc2−/− P11-12 pups to enter behavioral quiescence. n = 48 (Trpc2−/+) and 30 (Trpc2−/−). (k) Ultrasonic vocalizations emitted by isolated Trpc2−/+ or Trpc2−/− P13-14 pups, placed in a clean cage for 30 min at room temperature or with a heating pad underneath. Boxed portion quantified in (l). n = 22 (Trpc−/+), 21 (Trpc2−/−). Error bars show standard error of the mean. (l) Difference in the number of vocalizations between room temperature and heating pad conditions for each pup, at 20-30 min post-isolation (boxed portion in (k)). In all panels, boxplots show median, interquartile range, and 1.5 * interquartile range. For panels (g-l), p-value indicates results from two-sided t-test.