Extended Data Fig. 4: Morphological analysis and clustering of DP neurons.

a, General design of Cre-dependent MORF3 reporter mouse line²⁵. b, Lightsheet images and digital reconstructions of MORF-labelled DP projection neurons. These neurons exhibit detailed dendritic morphology. c, Reconstructed DP neurons coloured by distance of soma from the midline. Left: continuous colour gradient (blue = closest, red = farthest). Middle and right: neurons clustered into superficial (blue) and deep (red) groups based on distance-based classification. d, Left: classification of DP neurons into two morphological clusters: Cluster 1 neurons (blue) have smaller, less complex dendrites (average total dendritic length: 3024 ± 833 µm, average number of branches: 38 ± 12). Cluster 2 neurons (red) have larger, more complex dendrites (average total dendritic length: 5821 ± 958 µm, average number of branches: 76 ± 13). Right: differences between DP neurons with cell bodies in the superficial vs. deep layer. We initially measured the distance of each DP neuron’s cell body from the brain midline. Subsequently, we grouped all 47 neurons into two categories based on their distance from the midline (using k-means clustering method): (i) superficial (26 neurons, average distance of 288 ± 81 µm from the midline) and (ii) deep (21 neurons, average distance of 605 ± 106 µm from the midline). When comparing the superficial and deep clusters with the morphological clusters, the majority of deep layer DP neurons (18/21 neurons) fell within the smaller, less complex (deep layer average length: 3472 ± 1285 µm, average number of branches: 42 ± 16) morphological cluster, whereas the morphology of superficial DP neurons were significantly larger and more complex (average length: 4275 ± 1717 µm, average number of branches: 56 ± 24).