Original ResearchFull Report: Basic and Translational—Alimentary TractOptogenetic Demonstration of Functional Innervation of Mouse Colon by Neurons Derived From Transplanted Neural Cells
Section snippets
Mice
Wnt1::Cre mice26 were time plug−mated to Ai32(RCL-ChR2(H134R)/EYFP) mice (stock no: 012569; The Jackson Laboratory, Bar Harbor, ME), which express ChR2/EYFP fusion protein after exposure to Cre recombinase. Neurospheres were generated from the gut of embryonic (E)14.5 or postnatal (P)3−5 Wnt1::Cre;ChR2-EYFP mice. Wild-type C57BL/6 mice at 3−4 weeks of age were used as recipients.
Isolation and Culture of Enteric Neural Cells as Neurospheres
Enteric neurospheres were generated as described previously21 (see Supplementary Material for details). Neurospheres
Enteric Neural Cells Transplanted Into the Colon Generate Neurons That Project Axons to the Circular Muscle and Myenteric Ganglia of the Recipient
Enteric neural crest-derived cells expressing the light-sensitive ion channel ChR2 were isolated from E14.5 or P3−5 Wnt1::Cre;ChR2EYFP mice and grown in low attachment wells as described previously.21 Seven days after isolation from both the fetal or postnatal bowel, enteric neural cells formed neurospheres around 200 μm in diameter (Figure 1A). Neurospheres generated from E14.5 Wnt1::Cre;ChR2EYFP mice were termed fetal neurospheres and neurospheres generated from P3−5 Wnt1::Cre;ChR2EYFP mice
Discussion
Cell therapy offers the potential to treat diseases of the ENS.4, 9, 11 Significant progress has been made in the isolation of enteric neural cells from the bowel of humans and rodents, and in transplantation of cells into the bowel of rodents.7, 15, 17, 20, 22, 23, 37, 38, 39, 40 However, it had not been possible to determine whether transplanted cells generate different functional classes of enteric neurons, including interneurons and excitatory and inhibitory motor neurons, which are
Acknowledgments
The authors thank Annette Bergner and Jan Morgan for excellent technical assistance, and Vanda Lennon and Piers Emson for generously providing antisera.
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2022, Journal of Pediatric SurgeryCitation Excerpt :Subsequently, several investigators successfully isolated ENCDCs from embryonic and postnatal rodent gut [34–36] and demonstrated their survival up to 24 months following transplantation in mouse colon in vivo [37]. ENCDCs give rise to functioning neurons [38,39] that integrate into the neuronal circuitry of the host ENS in vivo [40]. Importantly, ENCDC transplantation was recently shown to ameliorate the decreased colonic motility seen in neuronal nitric oxide synthase (nNOS)-null mice four weeks after surgery [29].
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2021, Trends in Molecular MedicineEffects of optogenetic activation of the enteric nervous system on gastrointestinal motility in mouse small intestine
2020, Autonomic Neuroscience: Basic and ClinicalCitation Excerpt :Only recently has it been demonstrated that optogenetic technology can be used to effectively control the excitability of the enteric nervous system in vertebrates (Stamp et al., 2017; Hibberd et al., 2018; Spencer et al., 2019; Perez-Medina and Galligan, 2019).
Retinoic Acid Accelerates the Specification of Enteric Neural Progenitors from In-Vitro-Derived Neural Crest
2020, Stem Cell ReportsCitation Excerpt :Transplanted human cells populated both the myenteric and submucosal plexuses of the gut, demonstrating extensive migration within the gut wall and formation of neuronal networks with close interactions with the intact host ENS (Figure 4). Transplantation studies using postnatal human and murine endogenous enteric neural stem cells in mice demonstrated functional integration (Cooper et al., 2016, 2017; Stamp et al., 2017), and rescue of an enteric neuropathy (McCann et al., 2017). Transplanted hPSC-derived ENS progenitors, generated through dual-SMAD inhibition, integrate and migrate extensively within a mouse model of HSCR leading to increased survival (Fattahi et al., 2016).
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2020, Gastroparesis: Pathophysiology, Clinical Presentation, Diagnosis and Treatment
Conflicts of interest The authors disclose no conflicts.
Funding This work was supported by National Health and Medical Research Council (NHMRC) project grant APP1079234 to Heather M. Young, Lincon A. Stamp, and Joel C. Bornstein, and NHMRC Senior Fellowship APP1002506 to Heather M. Young. The study sponsor played no role in study design or in the collection, analysis, and interpretation of data.
Author names in bold designate shared co-first authorship.