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Abstract Quantitatively comparing brain-wide connectivity of different types of neuron is of vital importance in understanding the function of the mammalian cortex. Here we have designed an analytical approach to examine and compare datasets from hierarchical segmentation ontologies, and applied it to long-range presynaptic connectivity onto excitatory and inhibitory neurons, mainly located in layer 2/3 (L2/3), of mouse primary visual cortex (V1). We find that the origins of long-range connections onto these two general cell classes—as well as their proportions—are quite similar, in contrast to the inputs on to a cell type in L6. These anatomical data suggest that distal inputs received by the general excitatory and inhibitory classes of neuron in L2/3 overlap considerably.

Anatomical segmentation of high-resolution 3D microscopy datasets is necessary to map large samples at cellular resolution. Here the authors present a pipeline for automated mouse atlas propagation (aMAP) to segment fluorescence images of the adult mouse brain and validate it against human segmentations.