The Enteric Nervous System for Epithelial Researchers: Basic Anatomy, Techniques, and Interactions With the Epithelium

Kathleen T Walsh, Anne E Zemper, Kathleen T Walsh, Anne E Zemper

Abstract

The intestinal epithelium does not function in isolation, but interacts with many components including the Enteric Nervous System (ENS). Understanding ENS and intestinal epithelium interactions requires multidisciplinary approaches to uncover cells involved, mechanisms used, and the ultimate influence on intestinal physiology. This review is intended to serve as a reference for epithelial biologists interested in studying these interactions. With this in mind, this review aims to summarize the basic anatomy of the epithelium and ENS, mechanisms by which they interact, and techniques used to study these interactions. We highlight in vitro, ex vivo and in vivo techniques. Additionally, ENS influence on epithelial proliferation and gene expression within stem and differentiated cells as well as gastrointestinal cancer are discussed.

Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.

Figures

Figure 1
Figure 1
Anatomy of intestinal epithelium, ENS, and intestinal muscle layers. (A) Cartoon depiction of SI epithelium depicting villi with differentiated cell types and crypts with progenitor cell niche located above the ENS and musculature of intestine. (B) Cartoon depicting of colonic epithelium depicting differentiated cells and progenitors within crypts located above the ENS and musculature of intestine. (A, B) Mucosal glia located within mucosa, intraganglionic glia are located adjacent to neuronal cell bodies within ganglia, and intramuscular glia are located along nerves within muscle layers. Yellow nerves represent intrinsic nerves from submucosal plexus, orange nerves represent intrinsic nerves from myenteric plexus, and blue nerves represent extrinsic innervation into intestine. The legend is located below the graphic. (C) Immunofluorescent image of colonic epithelium. Epithelial cells (DAPI, blue) are located above submucosal and myenteric neurons. Neuronal nuclei marked with HuC/D (green) and neuronal projections marked with PGP9.5 (magenta). Note that the staining at the top of the crypts is background staining. (D) Longitudinal view of myenteric plexus depicting enteric neuron morphology using same markers as described in panel C. Circle represents one ganglia (yellow dashed circle). (C, D) Scale bar = 50 um. ICC, interstitial cells of Cajal.
Figure 2
Figure 2
Mechanisms of epithelial-ENS interactions and current techniques. (A) Paracrine model of neuron-epithelial cell communication. Diffusion of molecules between intestinal epithelial cells and enteric neurons may result in communication in both directions. Release of molecules (yellow or blue circles) from one cell can bind to receptors (red or magenta ovals) on the signal-receiving cell resulting in downstream signaling. (B) Depiction of direct synaptic connections between neurons and epithelial neuropod cells. Expression of synaptic proteins between the neuron and neuropod cell allows for direct communication between the two cells. As with diffusion, expression of hormones or neurotransmitters (yellow or blue circles) from the presynaptic cell binds to receptors (red or magenta ovals) on the opposing postsynaptic cell. Expression of pre- and postsynaptic proteins has been shown with this model. (C–H) Approaches used to study epithelial-ENS interactions using in vitro, ex vivo, or in vivo techniques: (C) optogenetics to manipulate ENS activity in vivo; (D) abdominal window to visualize intestinal activity in vivo in anesthetized animals; and (E) Ussing chamber to study secretion, signaling, or barrier function with ENS cultured on basal side. (The Ussing chamber can be an ex vivo or in vitro technique depending on type of epithelial tissue or cell monolayer used); (F) intestinal motility assay measures propagation of artificial fecal pellet in ex vivo setting; (G) co-culture of monolayers; or (H) organoids allowing for measurement of in vitro gene expression changes or barrier permeability with or without ENS influence on epithelium.

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