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Published May 11, 2022 | Submitted
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Single-cell profiling coupled with lineage analysis reveals distinct sacral neural crest contributions to the developing enteric nervous system


During development, the enteric nervous system (ENS) arises from neural crest cells that emerge from the neural tube, migrate to and along the gut, and colonize the entire intestinal tract. While much of the ENS arises from vagal neural crest cells that originate from the caudal hindbrain, there is a second contribution from the sacral neural crest that migrates from the caudal end of the spinal cord to populate the post-umbilical gut. By coupling single cell transcriptomics with axial-level specific lineage tracing in avian embryos, we compared the contributions between embryonic vagal and sacral neural crest cells to the ENS. The results show that the two neural crest populations form partially overlapping but also complementary subsets of neurons and glia in distinct ganglionic units. In particular, the sacral neural crest cells appear to be the major source of adrenergic/dopaminergic and serotonergic neurons, melanocytes and Schwann cells in the post-umbilical gut. In addition to neurons and glia, the results also reveal sacral neural crest contributions to connective tissue and mesenchymal cells of the gut. These findings highlight the specific properties of the sacral neural crest population in the hindgut and have potential implications for understanding development of the complex nervous system in the hindgut environment that may influence congenital neuropathies.

Additional Information

The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license. This version posted May 9, 2022. This work was supported by R01DE027568 and R35NS111564 to M.E.B. We thank Drs. Igor Antoshechkin and Vijaya Kumar and the Millard and Muriel Jacobs Genetics and Genomics Laboratory at California Institute of Technology for their guidance and support in bulk RNA-sequencing. We thank Jamie Tijerina and Rochelle Diamond from the Beckman Institute Flow Cytometry Facility for their help with the FACS. We thank Dr. Sisi Chen, Jeff Park, Prof. Matt Thomson and SPEC at Caltech for their dedicated support in optimization and guidance in single-cell RNA-sequencing. We thank Dr. Fan Gao and Bioinformatics Resource Center in the Beckman Institute at Caltech for guiding us through single-cell transcriptomic analysis. We appreciate the help from Prof. Carlos Lois for kindly sharing equipment with us to perform RIA concentration. We thank Dr. Michael L. Piacentino, Dr. Erica J. Hutchins and Prof. Angelike Stathopoulos for the helpful discussion on the manuscript. The authors have declared no competing interest.

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Submitted - 2022.05.09.491197v1.full.pdf


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