Fig. 5: Annotation of molecularly distinct glycinergic cell types in CN.

a Examples of reconstructed inhibitory neurons in a sagittal section, colored by cell type. b Example responses of inhibitory cells to current steps. c Left: UMAP visualization of morphological features of 116 inhibitory neurons (M-cluster), colored by expert cell type. Vertical: n = 38; SSC: n = 13; Cartwheel: n = 15; D-stellate: n = 15, L-stellate: n = 15. Right: Performance (Ave: 96.9%) of the classifier trained with morphological features. d Left: UMAP visualization of electrophysiological features along with anatomic locations (DCN:1; VCN:0) of 172 inhibitory neurons (E-cluster). Vertical: n = 50, SSC: n = 21; cartwheel: n = 21; D-stellate: n = 26, L-stellate: n = 53. Right: Performance (Ave: 97.9%) of the classifier trained with electrophysiological features and anatomic locations. See Fig. S4e, f for more analysis. e Left: UMAP visualization of 94 Patch-seq inhibitory neurons, colored by T-clusters. Right: Proportion of Patch-seq cells in each T-cluster assigned to a specific cell type. T-cluster 1: 12/12 are cartwheel cells; T-cluster 2: 20/21 are D-stellate cells; T-cluster 3: 35/35 are L-stellate cells; T-cluster 4: 10/12 are SSCs; T-cluster 5: 12/14 are vertical cells. f Top: 94 Patch-seq inhibitory neurons mapped onto snRNA-seq UMAP space, colored by T-cluster. Bottom: Proportion of Patch-seq cells assigned to each T-cluster in (e) mapped onto one of 13 distinct clusters in UMAP space. g UMAP visualization of all CN neuronal clusters, along with annotation with morpho-electrophysiological types. h UMAP visualization and annotation of all glycinergic CN cell types, and normalized expression of their respective marker genes. i FISH co-staining for Slc6a5/Stac, or for Slc6a5/Penk in sagittal sections. Lines indicate density of double-labeled neurons along two CN axes. Insets show proportion of double-labeled cells among all Stac⁺ or Penk⁺ cells counted across eight or six sections. j FISH co-staining for Slc6a5/tdTomato (left), or for Slc17a6/tdTomato (right) in sagittal sections in Penk-Cre:Ai9 mice. Insets show proportion of double-labeled cells among all tdTomato⁺ cells counted across six sections. k Micrographs showing labeled cells for recording (left), example responses to current steps (middle), and morphologies of labeled neurons (right) in Penk-Cre:Ai9 mice. l Distribution of labeled cells (red) in electrophysiological or morphological UMAP space of all inhibitory neurons. Source data are provided in the Source Data file.