Transcription and Signaling Regulators in Developing Neuronal Subtypes of Mouse and Human Enteric Nervous System

Fatima Memic,Viktoria Knoflach,Khomgrit Morarach,Rebecca Sadler,Catia Laranjeira,Jens Hjerling-Leffler,Erik Sundström,Vassilis Pachnis,Ulrika Marklund

doi:10.1053/j.gastro.2017.10.005

Fatima Memic, Viktoria Knoflach + Show 7 more

Open Access

https://doi.org/10.1053/j.gastro.2017.10.005

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Abstract

Background & AimsThe enteric nervous system (ENS) regulates gastrointestinal function via different subtypes of neurons, organized into fine-tuned neural circuits. It is not clear how cell diversity is created within the embryonic ENS; information required for development of cell-based therapies and models of enteric neuropathies. We aimed to identify proteins that regulate ENS differentiation and network formation.MethodsWe generated and compared RNA expression profiles of the entire ENS, ENS progenitor cells, and non-ENS gut cells of mice, collected at embryonic days 11.5 and 15.5, when different subtypes of neurons are formed. Gastrointestinal tissues from R26ReYFP reporter mice crossed to Sox10-CreERT2 or Wnt1-Cre mice were dissected and the 6 populations of cells were isolated by flow cytometry. We used histochemistry to map differentially expressed proteins in mouse and human gut tissues at different stages of development, in different regions. We examined enteric neuronal diversity and gastric function in Wnt1-Cre x Sox6fl/fl mice, which do not express the Sox6 gene in the ENS.ResultsWe identified 147 transcription and signaling factors that varied in spatial and temporal expression during development of the mouse ENS. Of the factors also analyzed in human ENS, most were conserved. We uncovered 16 signaling pathways (such as fibroblast growth factor and Eph/ephrin pathways). Transcription factors were grouped according to their specific expression in enteric progenitor cells (such as MEF2C), enteric neurons (such as SOX4), or neuron subpopulations (such as SATB1 and SOX6). Lack of SOX6 in the ENS reduced the numbers of gastric dopamine neurons and delayed gastric emptying.ConclusionsUsing transcriptome and histochemical analyses of the developing mouse and human ENS, we mapped expression patterns of transcription and signaling factors. Further studies of these candidate determinants might elucidate the mechanisms by which enteric stem cells differentiate into neuronal subtypes and form distinct connectivity patterns during ENS development. We found expression of SOX6 to be required for development of gastric dopamine neurons.

Highlights

BACKGROUND & AIMSThe enteric nervous system (ENS) regulates gastrointestinal function via different subtypes of neurons, organized into fine-tuned neural circuits
As SOX10 marks dividing ENS progenitor cells,[8] we could retrieve this subpopulation by inducing reporter expression in Sox10CreERT2 Â R26ReYFP embryos.[6]
Transcriptomes of YFPÀ cells, derived from Wnt1Cre Â R26ReYFP gut at E11.5 and E15.5 representing non-ENS gut tissue were included in the analysis (Figure 1A)

Summary

Introduction

The enteric nervous system (ENS) regulates gastrointestinal function via different subtypes of neurons, organized into fine-tuned neural circuits. It is not clear how cell diversity is created within the embryonic ENS; information required for development of cell-based therapies and models of enteric neuropathies. CONCLUSIONS: Using transcriptome and histochemical analyses of the developing mouse and human ENS, we mapped expression patterns of transcription and signaling factors. Further studies of these candidate determinants might elucidate the mechanisms by which enteric stem cells differentiate into neuronal subtypes and form distinct connectivity patterns during ENS development. The diverse types of neurons of the enteric nervous system (ENS) are required for normal gastrointestinal functions, yet how cellular diversity is created during ENS development remains unknown

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