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

doi:10.1016/j.isci.2023.107070

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^*

^*Corresponding author for this work

Erasmus University Medical Centre
Emory University
Sophia Children's Hospital

Research output: Contribution to journal › Article › Academic › peer-review

17 Citations (Scopus)

108 Downloads (Pure)

Abstract

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.

Original language	English
Article number	107070
Number of pages	17
Journal	iScience
Volume	26
Issue number	7
DOIs	https://doi.org/10.1016/j.isci.2023.107070
Publication status	Published - 21 Jul 2023

Bibliographical note

Funding Information:
We want to thank Remco Hoogenboezem for mapping the scRNA-seq data, Emma de Pater for useful discussions, Soheil Yousefi for bioinformatics assistance, and the optical imaging center (OIC) of the Erasmus University Medical Center for assistance with confocal microscopy. Graphical abstract was created with BioRender.com. This work was funded by the Friends of the Sophia Foundation ( SSWO WAR-63 ).

Publisher Copyright:
© 2023 The Author(s)

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PIIS2589004223011471Final published version, 6.62 MBLicence: CC BY

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title = "Unbiased characterization of the larval zebrafish enteric nervous system at a single cell transcriptomic level",

abstract = "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.",

author = "Kuil, \{Laura E.\} and Kakiailatu, \{Naomi J.M.\} and Windster, \{Jonathan D.\} and Eric Bindels and Zink, \{Joke T.M.\} and \{van der Zee\}, Gaby and Hofstra, \{Robert M.W.\} and Shepherd, \{Iain T.\} and Veerle Melotte and Alves, \{Maria M.\}",

note = "Funding Information: We want to thank Remco Hoogenboezem for mapping the scRNA-seq data, Emma de Pater for useful discussions, Soheil Yousefi for bioinformatics assistance, and the optical imaging center (OIC) of the Erasmus University Medical Center for assistance with confocal microscopy. Graphical abstract was created with BioRender.com. This work was funded by the Friends of the Sophia Foundation ( SSWO WAR-63 ). Publisher Copyright: {\textcopyright} 2023 The Author(s)",

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AU - Bindels, Eric

AU - Zink, Joke T.M.

AU - van der Zee, Gaby

AU - Hofstra, Robert M.W.

AU - Shepherd, Iain T.

AU - Melotte, Veerle

AU - Alves, Maria M.

N1 - Funding Information: We want to thank Remco Hoogenboezem for mapping the scRNA-seq data, Emma de Pater for useful discussions, Soheil Yousefi for bioinformatics assistance, and the optical imaging center (OIC) of the Erasmus University Medical Center for assistance with confocal microscopy. Graphical abstract was created with BioRender.com. This work was funded by the Friends of the Sophia Foundation ( SSWO WAR-63 ). Publisher Copyright: © 2023 The Author(s)

PY - 2023/7/21

Y1 - 2023/7/21

N2 - 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.

AB - 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.

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JO - iScience

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Unbiased characterization of the larval zebrafish enteric nervous system at a single cell transcriptomic level

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