Unbiased characterization of the larval zebrafish enteric nervous system at a single cell transcriptomic level
Laura E. Kuil,
Naomi J.M. Kakiailatu,
Jonathan D. Windster,
Eric Bindels,
Joke T.M. Zink,
Gaby van der Zee,
Robert M.W. Hofstra,
Iain T. Shepherd,
Veerle Melotte,
Maria M. Alves
Affiliations
Laura E. Kuil
Department of Clinical Genetics, Erasmus University Medical Center, Sophia Children’s Hospital, Rotterdam, the Netherlands; Corresponding author
Naomi J.M. Kakiailatu
Department of Clinical Genetics, Erasmus University Medical Center, Sophia Children’s Hospital, Rotterdam, the Netherlands
Jonathan D. Windster
Department of Clinical Genetics, Erasmus University Medical Center, Sophia Children’s Hospital, Rotterdam, the Netherlands; Department of Pediatric Surgery, Erasmus University Medical Center, Sophia Children’s Hospital, Rotterdam, the Netherlands
Eric Bindels
Department of Hematology, Erasmus MC, Rotterdam, the Netherlands
Joke T.M. Zink
Department of Hematology, Erasmus MC, Rotterdam, the Netherlands
Gaby van der Zee
Department of Clinical Genetics, Erasmus University Medical Center, Sophia Children’s Hospital, Rotterdam, the Netherlands
Robert M.W. Hofstra
Department of Clinical Genetics, Erasmus University Medical Center, Sophia Children’s Hospital, Rotterdam, the Netherlands
Iain T. Shepherd
Department of Biology, Emory University, Atlanta, GA, USA
Veerle Melotte
Department of Clinical Genetics, Erasmus University Medical Center, Sophia Children’s Hospital, Rotterdam, the Netherlands; Department of Pathology, GROW-School for Oncology and Developmental Biology, Maastricht University Medical Center, Maastricht, the Netherlands
Maria M. Alves
Department of Clinical Genetics, Erasmus University Medical Center, Sophia Children’s Hospital, Rotterdam, the Netherlands; Corresponding author
Summary: The enteric nervous system (ENS) regulates many gastrointestinal functions including peristalsis, immune regulation and uptake of nutrients. Defects in the ENS can lead to severe enteric neuropathies such as Hirschsprung disease (HSCR). Zebrafish have proven to be fruitful in the identification of genes involved in ENS development and HSCR pathogenesis. However, composition and specification of enteric neurons and glial subtypes at larval stages, remains mainly unexplored. Here, we performed single cell RNA sequencing of zebrafish ENS at 5 days post-fertilization. We identified vagal neural crest progenitors, Schwann cell precursors, and four clusters of differentiated neurons. In addition, a previously unrecognized elavl3+/phox2bb-population of neurons and cx43+/phox2bb-enteric glia was found. Pseudotime analysis supported binary neurogenic branching of ENS differentiation, driven by a notch-responsive state. Taken together, we provide new insights on ENS development and specification, proving that the zebrafish is a valuable model for the study of congenital enteric neuropathies.
