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The orbitofrontal cortex (OFC) is involved in diverse brain functions via its extensive projections to multiple target regions. There is a growing understanding of the overall outputs of the OFC at the population level, but reports of the projection patterns of individual OFC neurons across different cortical layers remain rare. Here, by combining neuronal sparse and bright labeling with a whole-brain florescence imaging system (fMOST), we obtained an uninterrupted three-dimensional whole-brain dataset and achieved the full morphological reconstruction of 25 OFC pyramidal neurons. We compared the whole-brain projection targets of these individual OFC neurons in different cortical layers as well as in the same cortical layer. We found cortical layer-dependent projections characterized by divergent patterns for information delivery. Our study not only provides a structural basis for understanding the principles of laminar organizations in the OFC, but also provides clues for future functional and behavioral studies on OFC pyramidal neurons.

Figure 1. Strategy for sparse neuronal labeling with strong brightness.
 

To obtain brain-wide projections of OFC populations, the authors sparsely and brightly labeled neurons in the OFC in mice and performed whole-brain imaging using the dual-color fluorescence micro-optical sectioning tomography (fMOST) system. The study not only facilitates our understanding of the laminar organization of the OFC but also provides detailed anatomical clues for further functional and behavior studies related to OFC pyramidal neurons.

 
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