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

Kuil, Laura E.; Kakiailatu, Naomi; Windster, Jonathan D; Bindels, Eric M.; Peulen-Zink, Joke; Zee, G. van der; Hofstra, Robert; Shepherd, Iain T.; Melotte, Veerle; Alves, Maria M.

doi:10.1016/j.isci.2023.107070

Cited by 8 publications

(2 citation statements)

References 66 publications

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“…Once reaching their destinations along the gut, which can extend from the foregut to the hindgut, ENCCs spatially pattern along the gut circumference, and begin to differentiate into enteric neurons [ 31 , 35 ] ( Figure 1C ), or glia [ 40 , 41 ], between 54 hpf and 7 days post fertilization (dpf) during the larval stage. Differentiating neurons express enteric neuronal subtype markers, such as vipb , nos1 , chata , and pbx3b [ 36 , 41–43 ], while cells with differentiating glial identity express the markers her4 , cx43 , and s100b [ 40 , 41 ]. The zebrafish ENS continues neurogenesis after 7 dpf [ 40 , 43 ]; however, the ENS already contains an identifiable neuropil structure by 7 dpf [ 44 ].…”

Section: Zebrafish Ens Developmental Stages and Major Eventsmentioning

confidence: 99%

Genetic regulation of enteric nervous system development in zebrafish

Uribe

2024

Biochemical Society Transactions

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The enteric nervous system (ENS) is a complex series of interconnected neurons and glia that reside within and along the entire length of the gastrointestinal tract. ENS functions are vital to gut homeostasis and digestion, including local control of peristalsis, water balance, and intestinal cell barrier function. How the ENS develops during embryological development is a topic of great concern, as defects in ENS development can result in various diseases, the most common being Hirschsprung disease, in which variable regions of the infant gut lack ENS, with the distal colon most affected. Deciphering how the ENS forms from its progenitor cells, enteric neural crest cells, is an active area of research across various animal models. The vertebrate animal model, zebrafish, has been increasingly leveraged to understand early ENS formation, and over the past 20 years has contributed to our knowledge of the genetic regulation that underlies enteric development. In this review, I summarize our knowledge regarding the genetic regulation of zebrafish enteric neuronal development, and based on the most current literature, present a gene regulatory network inferred to underlie its construction. I also provide perspectives on areas for future zebrafish ENS research.

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Section: Zebrafish Ens Developmental Stages and Major Eventsmentioning

confidence: 99%

Genetic regulation of enteric nervous system development in zebrafish

Uribe

2024

Biochemical Society Transactions

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“…Recently, integration of single-cell RNA sequencing (scRNA-seq) into the zebrafish model for exploration of ENS development has yielded invaluable insights into what genes are expressed during ENS developmental phases [ 20 , 21 ]. Concurrently, the successful adoption of CRISPR technology for gene disruption in zebrafish, known for its high efficiency in analyzing phenotypes directly in the injected generation (F0), also known as “crispants”, facilitates swift assessment of candidate genes [ 22 , 23 ].…”

Section: Introductionmentioning

confidence: 99%

A targeted CRISPR-Cas9 mediated F0 screen identifies genes involved in establishment of the enteric nervous system

Moreno-Campos,

Singleton,

Uribe

2024

PLoS ONE

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The vertebrate enteric nervous system (ENS) is a crucial network of enteric neurons and glia resident within the entire gastrointestinal tract (GI). Overseeing essential GI functions such as gut motility and water balance, the ENS serves as a pivotal bidirectional link in the gut-brain axis. During early development, the ENS is primarily derived from enteric neural crest cells (ENCCs). Disruptions to ENCC development, as seen in conditions like Hirschsprung disease (HSCR), lead to the absence of ENS in the GI, particularly in the colon. In this study, using zebrafish, we devised an in vivo F0 CRISPR-based screen employing a robust, rapid pipeline integrating single-cell RNA sequencing, CRISPR reverse genetics, and high-content imaging. Our findings unveil various genes, including those encoding opioid receptors, as possible regulators of ENS establishment. In addition, we present evidence that suggests opioid receptor involvement in the neurochemical coding of the larval ENS. In summary, our work presents a novel, efficient CRISPR screen targeting ENS development, facilitating the discovery of previously unknown genes, and increasing knowledge of nervous system construction.

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