Intestinal Nerve Research Articles

The Pathological Mechanism Between the Intestine and Brain in the Early Stage of Parkinson's Disease.

Parkinson's disease (PD) is the second most common chronic progressive neurodegenerative disease. The main pathological features are progressive degeneration of neurons and abnormal accumulation of α-synuclein. At present, the pathogenesis of PD is not completely clear, and many changes in the intestinal tract may be the early pathogenic factors of PD. These changes affect the central nervous system (CNS) through both nervous and humoral pathways. α-Synuclein deposited in the intestinal nerve migrates upward along the vagus nerve to the brain. Inflammation and immune regulation mediated by intestinal immune cells may be involved, affecting the CNS through local blood circulation. In addition, microorganisms and their metabolites may also affect the progression of PD. Therefore, paying attention to the multiple changes in the intestinal tract may provide new insight for the early diagnosis and treatment of PD.

Large abdominal cyst of the fetus: A rare diagnosis of neonatal megaduodenum

We present a rare case of neonatal duodenal malformation. Herein, we provide a differential diagnosis for a large abdominal cyst in a prenatal fetus and explore the possible causes and treatment of neonatal megaduodenum. We retrospectively analyzed one case of megaduodenum from fetal examination to neonatal diagnosis and treatment. Ultrasonography of the fetus at 12 weeks' gestation revealed a cystic mass in the abdominal cavity, measuring about 0.7 cm × 0.5 cm × 0.4 cm. The cystic mass grew in equal proportion to the fetus. At 32 weeks’ gestation, the peritoneal cystic mass was enlarged to 9.0 cm × 7.9 cm × 5.8 cm, and the mass was found to be communicating with the stomach. Intraoperatively, the mass was about 10-cm long and 6 cm in diameter. The duodenal papilla was located about 1.5 cm above the proximal junction. The megaduodenum was removed, and the distal and proximal intestines were anastomosed end-to-end. The patient recovered smoothly postoperatively and gained 550 g 1 month after discharge. The results of amniotic fluid high-throughput sequencing identified Seq [Grch37] del (16) (p12.2) Chr16: g.22000000_22440000del. There was a copy number deletion of about 0.44 Mb on the short arm of chromosome 16, which was suspected to be pathogenic and involved seven protein-coding genes. Histopathologically, in the same paraffin specimen, there was a mixture of stunted ganglion cells, ganglion dysplasia, and well-developed ganglion cells. Neonatal megaduodenum was considered as a differential diagnosis of the large cystic mass in the fetal abdomen. Further additional evidence was needed to prove whether the mutation was pathogenic, which also suggested that the etiology is related to genetic variation. The nerve plexus and ganglion cells were poorly developed, which further supported that the intestinal nerve abnormality was the etiology. Excision was safe and effective for treating megaduodenum.

5-hydroxytryptamine produced by enteric serotonergic neurons initiates colorectal cancer stem cell self-renewal and tumorigenesis

Colorectal cancer stem cells (CSCs) contribute to colorectal tumorigenesis and metastasis. Colorectal CSCs reside within specialized niches and harbor self-renewal and differentiation capacities. However, the niche regulations of CSCs remain unclear. Here, we show that intestinal nerve cells are required for CSC self-renewal and colorectal tumorigenesis. Enteric serotonergic neurons produce 5-hydroxytryptamine (5-HT) to function as a modulator of CSC self-renewal. 5-HT receptors HTR1B/1D/1F are highly expressed in colorectal CSCs and engage with 5-HT to initiate Wnt/β-catenin signaling. Mechanistically, colorectal cancer (CRC)-enriched microbiota metabolite isovalerate suppresses the enrichment of the NuRD complex onto Tph2 promoter to initiate Tph2 expression, leading to 5-HT production. 5-HT signaling is correlated with CRC severity. Blocking 5-HT signaling in mice not only inhibits the self-renewal of colorectal CSCs but also displays therapeutic efficacy against CRC tumors. Our findings reveal a cross talk between intestinal neurons and tumor cells that serves as an additional layer for CSC regulation.

Gut microbiota drives macrophage-dependent self-renewal of intestinal stem cells via niche enteric serotonergic neurons.

Lgr5+ intestinal stem cells (ISCs) reside within specialized niches at the crypt base and harbor self-renewal and differentiation capacities. ISCs in the crypt base are sustained by their surrounding niche for precise modulation of self-renewal and differentiation. However, how intestinal cells in the crypt niche and microbiota in enteric cavity coordinatelyregulate ISC stemness remains unclear. Here, we show that ISCs are regulated by microbiota and niche enteric serotonergic neurons. Thegut microbiota metabolite valeric acid promotes Tph2 expression in enteric serotonergic neurons via blocking the recruitment of the NuRD complex onto Tph2 promoter. 5-hydroxytryptamine (5-HT) in turnactivates PGE2 production in aPGE2+ macrophage subset through its receptors HTR2A/3 A; and PGE2 via binding its receptors EP1/EP4,promotes Wnt/β-catenin signaling in ISCs to promote theirself-renewal. Our findings illustrate a complex crosstalk among microbiota, intestinal nerve cells, intestinal immune cells and ISCs, revealing a new layer of ISC regulation by niche cells and microbiota.

A Review on the Marek's Disease Outbreak and Its Virulence-Related meq Genovariation in Asia between 2011 and 2021.

Simple SummaryMarek’s disease is continuously causing an economic loss in Asia, despite the wide use of vaccines in the last decade. This review aims at summarizing the outbreak, the virulence-related meq gene variation, and the pathological information of Marek’s disease in the last decade in Asia. We found that a total of 132 viral strains emerged in 12 countries with different meq sequences. Among the evidence we have collected, 12 strains found in China were vaccine-resistant, reaching a mortality rate of 30% and above. This evidence requires the related region in China to consider the renewal of its vaccination type; however, more studies regarding the vaccination efficiency in other Asian countries are recommended, as the current information is not enough. The visceral tumor is the most common pathological type (13 in 16 studies) in Asia, while it is possible that a neural type may exist. We suggest that farmers monitor the behavioral changes of chickens to identify this harmful disease at the early stage. The phylogenetic analysis shows interconnection between Middle Eastern, South Asian, and East Asian countries that are geologically connected—poultry trading managers should consider the potential of viral transmitting.Marek’s disease is an infectious disease in poultry that usually appears in neural and visceral tumors. This disease is caused by Gallid alphaherpesvirus 2 infection in lymphocytes, and its meq gene is commonly used in virulent studies for coding the key protein functional in oncogenic transformation of the lymphocytes. Although vaccines have been introduced in many countries to control its spread and are proven to be efficient, recent records show a decline of such efficiency due to viral evolution. In this study, we reviewed the outbreak of Marek’s disease in Asia for the last 10 years, together with associated meq sequences, finding a total of 36 studies recording outbreaks with 132 viral strains in 12 countries. The visceral type is the most common (13 in 16 studies) form of Marek’s disease, but additional unobserved neural changes may exist. MD induces liver lymphoma most frequently (11 in 14 studies), and tumors were also found in spleen, kidney, heart, gizzard, skin, intestine, lung, and sciatic nerve. Twelve viral strains distributed in China have been reported to escape the CVI988 vaccine, reaching a mortality rate of more than 30%. Phylogenetic analyses show the internal connection between the Middle East (Turkey, Iraq, Iran, Saudi Arabia), South Asia (India, Indonesia), and East Asia (China and Japan), while external viral communications might occasionally occur. In 18 strains with both sequential and mortality data, amino acid alignment showed several point substitutions that may be related to its virulence. We suggest more behavioral monitoring in Marek’s disease-endemic regions and further studies on strain virulence, together with its Meq protein structural changes.

Diagnosis of Marek’s Disease through Histological, Immunohisto-Chemical and Molecular Applications: A Comparative Study

A total of 615 dead birds from various commercial poultry farms in and around Tirupati of Chittoor district of Andhra Pradesh were examined and among them 63 birds (10.24%) revealed suspected lesions for Marek’s disease. The gross and microscopic examination of the different tissues was carried out and further confirmed as Marek’s disease by immunohistochemical and molecular applications. Histologically, infiltration of the pleomorphic lymphoid cells was observed in the liver, heart, lung, kidney, ovary, spleen, thymus, pancreas, proventriculus, intestine, skin, skeletal muscle and sciatic nerve. Immunohistochemical staining revealed mild to moderate positive reactivity for MDV antigens in the cytoplasm of the infiltrating lymphocytes in all the organs. Mild positive reactivity for Marek’s disease was detected in the hepatocytes, tubular epithelial cells of the kidney, myocardial fibers and epithelial cells of the intestine. Molecular detection of the Meq gene of serotype 1 MDV in the tissue samples revealed an expected size of 1081 bp in PCR products in Marek’s disease - positive birds. The sequence showed 99.7% homology with that of different strains of serotype-1 specific Gallid Herpes virus 2 of Kerala and Ludhiana. The above studies indicated that the incidence of Marek’s disease among the vaccinated chicken flock was evident. The immunohistochemical studies revealed the distribution pattern of the Marek’s disease virus in internal organs and the PCR product was amplified on histologically diagnosed tissues.

Development of Human Gut Organoids With Resident Tissue Macrophages as a Model of Intestinal Immune Responses.

Development of Human Gut Organoids With Resident Tissue Macrophages as a Model of Intestinal Immune Responses.

Pathological changes in internal organs and intestines with colidispensions in young children

The aim of this work was to study and atologoanatomical changes in internal organs, the brain and intestines, the serotypes of Escherichia coli, its localization, and the state of the intestinal intramural nerves. The corpses of 40 children from the neonatal period to 1.5 years of age were examined (in the first week of life - 3, in the second - 3, from 1 to 3 months of age - 13, from 3 to 6 months - 13, from 6 months to 1 years - 6, from 1 year to 1.5 years - 2).

Loss of Integrin-Linked Kinase Leads to Dysplasia of the Colon in Mice

Normal development of the colon is essential for well-being during the life span. Integrin linked kinase (ILK), which is a key part of the integrin signaling pathway, connects intracellular and extracellular signaling pathways and is evolutionally conserved. It has been shown that ILK knockouts can lead to abnormal formation of various organs. In our study, we found that deletion of ILK by platelet-derived growth factor receptor B (PDGFR[Formula: see text]-Cre leads to the abnormal migration of neural crest cells (NCCs) and the shortening of the gastrointestinal tract, along with the expansion of the colon cavity and dysplasia of the intestinal nerve fibers. In conclusion, ILK is required for normal colon development.

Regulation of the Autonomic Nervous System on Intestine.

Intestine is composed of various types of cells including absorptive epithelial cells, goblet cells, endocrine cells, Paneth cells, immunological cells, and so on, which play digestion, absorption, neuroendocrine, immunological function. Intestine is innervated with extrinsic autonomic nerves and intrinsic enteric nerves. The neurotransmitters and counterpart receptors are widely distributed in the different intestinal cells. Intestinal autonomic nerve system includes sympathetic and parasympathetic nervous systems, which regulate cellular proliferation and function in intestine under physiological and pathophysiological conditions. Presently, distribution and functional characteristics of autonomic nervous system in intestine were reviewed. How autonomic nervous system regulates intestinal cell proliferation was discussed. Function of autonomic nervous system on intestinal diseases was extensively reviewed. It might be helpful to properly manipulate autonomic nervous system during treating different intestinal diseases.

Renal Denervation Attenuates Neuroinflammation in the Brain by Regulating Gut-Brain Axis in Rats With Myocardial Infarction.

Aims: The development of neuroinflammation deteriorates the prognosis of myocardial infarction (MI). We aimed to investigate the effect of renal denervation (RDN) on post-MI neuroinflammation in rats and the related mechanisms.Methods and Results: Male adult Sprague-Dawley rats were subjected to sham or ligation of the left anterior descending coronary artery to induce MI. One week later, the MI rats received a sham or RDN procedure. Their cardiac functions were analyzed by echocardiography, and their intestinal structures, permeability, and inflammatory cytokines were tested. The intestinal microbiota were characterized by 16S rDNA sequencing. The degrees of neuroinflammation in the brains of rats were analyzed for microglia activation, inflammatory cytokines, and inflammation-related signal pathways. In comparison with the Control rats, the MI rats exhibited impaired cardiac functions, intestinal injury, increased intestinal barrier permeability, and microbial dysbiosis, accompanied by increased microglia activation and pro-inflammatory cytokine levels in the brain. A RDN procedure dramatically decreased the levels of renal and intestinal sympathetic nerve activity, improved cardiac functions, and mitigated the MI-related intestinal injury and neuroinflammation in the brain of MI rats. Interestingly, the RDN procedure mitigated the MI-increased intestinal barrier permeability and pro-inflammatory cytokines and plasma LPS as well as ameliorated the gut microbial dysbiosis in MI rats. The protective effect of RDN was not significantly affected by treatment with intestinal alkaline phosphatase but significantly reduced by L-phenylalanine treatment in MI rats.Conclusions: RDN attenuated the neuroinflammation in the brain of MI rats, associated with mitigating the MI-related intestinal injury.

Therapeutic effect and mechanism of Shouhui Tongbian Capsules on slow transit constipation model mice

Shouhui Tongbian Capsules was used to explore the therapeutic effect and potential mechanism on slow transit constipation model mice induced by loperamide hydrochloride. In the experiment, loperamide hydrochloride-induced ICR mice were used as the model of slow transit constipation. Fifty ICR mice were divided into the blank group, model group and high, medium and low dose groups of Shouhui Tongbian Capsules extract(100, 200 and 400 mg·kg~(-1)). The model group and the administration groups were then modeled using loperamide hydrochloride intragastrically to obtain slow transit constipation. After successful modeling, high, medium and low doses of drugs were given to each drug group by intragastric administration. After 14 days of administration, the first defecation time, 6 h defecation grain number, 6 h defecation wet weight and dry weight, black feces discharged within 6 h and the fecal water content were measured. Intestinal tissues were taken for c-Kit and SCF immunohistochemical sections to detect the expression of c-Kit and SCF in the blank group, model group and high, medium and low dose groups of the medicinal extract of Shouhui Tongbian Capsules. The tissue changes in the intestinal wall of mice were detected by HE staining. At the same time, partial intestinal tissues were taken to test the activity of ATP synthase and isocitrate dehydrogenase in intestinal tissues of mice. RESULTS:: showed that Shouhui Tongbian Capsules effectively improved the symptoms of slow transit constipation in ICR mice and promoted intestinal movement. Shouhui Tongbian Capsules obviously shortened the time of discharging black stool for the first time, improved the intestinal propulsion rate, increased the water content and amount of feces, and improved the constipation symptoms. Mechanism study revealed that Shouhui Tongbian Capsules increased ATP synthase activity and mitochondrial isocitrate dehydrogenase activity in intestinal tissue, and up-regulated c-Kit/SCF signaling pathway to promote interstitial Cajal cells proliferation, intestinal nerve transmission, intestinal motility and transport capacity.

On the question of the peripheral innervation of the rectum. Critical and experimental research

All experiments were made by you 56. Of these, 10 experiments with negative pressure, 8 experiments with double registration (according to Courtade and Guyon` y), 15 with simultaneous registration of three bowel sections (colonis, recti and sphinct. Inter.), 4 experiments with irritation spinal roots, 2 experiments for examining the external sphincter. The remaining 17 embrace the usual experiments of investigating the movements of the recti under the influence of the irritated intestinal nerves according to the method indicated by us in the beginning of our work; this also included 5 experiments with combined irritations n. erigentis and n. hypogastrici.

Simulations of Myenteric Neuron Dynamics in Response to Mechanical Stretch.

Background Intestinal sensitivity to mechanical stimuli has been studied intensively in visceral pain studies. The ability to sense different stimuli in the gut and translate these to physiological outcomes relies on the mechanosensory and transductive capacity of intrinsic intestinal nerves. However, the nature of the mechanosensitive channels and principal mechanical stimulus for mechanosensitive receptors are unknown. To be able to characterize intestinal mechanoelectrical transduction, that is, the molecular basis of mechanosensation, comprehensive mathematical models to predict responses of the sensory neurons to controlled mechanical stimuli are needed. This study aims to develop a biophysically based mathematical model of the myenteric neuron with the parameters constrained by learning from existing experimental data. Findings. The conductance-based single-compartment model was selected. The parameters in the model were optimized by using a combination of hand tuning and automated estimation. Using the optimized parameters, the model successfully predicted the electrophysiological features of the myenteric neurons with and without mechanical stimulation. Conclusions The model provides a method to predict features and levels of detail of the underlying physiological system in generating myenteric neuron responses. The model could be used as building blocks in future large-scale network simulations of intrinsic primary afferent neurons and their network.

Vasoactive Intestinal Polypeptide (VIP) in the Intestinal Mucosal Nerve Fibers in Dogs with Inflammatory Bowel Disease.

Simple SummaryCanine inflammatory bowel disease (IBD)—a group of gastrointestinal disorders—is a serious problem in veterinary medicine. The etiology of IBD remains unknown, and its diagnosis and effective treatment are difficult. One of the less-known aspects of IBD pathology is the influence of this disease on the enteric nervous system, which is located in the intestinal wall and regulates most of the gastrointestinal functions. Therefore, the aim of the present study was to evaluate the influence of IBD on the intramucosal nerve fibers containing vasoactive intestinal polypeptide (VIP). VIP is one of the most important substances produced by the enteric nervous structures that is involved in many regulatory processes in the gastrointestinal tract. The obtained results show that IBD induces changes in the density of intramucosal VIP-positive nerve fibers in the canine gastrointestinal tract. It suggests that VIP is involved in the pathological processes occurring during this disease. Observed changes may be a result of neuroprotective and/or adaptive processes regulated by VIP, aimed at the homeostasis maintenance in the inflamed gastrointestinal (GI) tract and induced by proinflammatory factors.Canine inflammatory bowel disease (IBD) is a group of enteropathies with nonspecific chronic symptoms and poorly understood etiology. Many aspects connected with IBD are not understood. One of them is the participation of the intestinal nervous system in the development of pathological processes. Thus, this study aimed to demonstrate changes in the density of intramucosal nerve fibers containing vasoactive intestinal polypeptide (VIP)—one of the most important intestinal nervous factors caused by the various stages of IBD development. Mucosal biopsy specimens collected from the duodenum, jejunum and descending colon of healthy dogs and dogs with varied severity of IBD were included in the experiment. The density of VIP-like immunoreactive (VIP-LI) nerves was determined by a single immunofluorescence technique and a semi-quantitative method consisting in VIP-LI fiber counts in the field of view (0.1 mm2). The obtained results indicate that IBD induces changes in the density of mucosal VIP-LI nerve fibers in the canine gastrointestinal tract. The initial decrease is followed by an increase in VIP-like immunoreactivity in successive stages of the disease. These observations show that VIP is a neuronal factor that participates in the pathological processes connected with canine IBD. The observed changes probably result from the neuroprotective and/or adaptive properties of VIP. Protective and adaptive reactions induced by inflammation aim to protect the GI tract against damage by proinflammatory factors and ensure the homeostasis in the enteric nervous system (ENS) under the conditions changed by the disease process.

Chronic high fat diet impairs glucagon like peptide-1 sensitivity in vagal afferents

Dysfunction of the gut-brain axis is one of the potential contributors to the pathophysiology of obesity and is therefore a potential target for treatment. Vagal afferents innervating the gut play an important role in controlling energy homeostasis. There is an increasing evidence for the role of vagal afferents in mediating the anorexigenic effects of glucagon-like peptide-1 (GLP-1), an important satiety and incretin hormone. This study aimed to examine the effect of chronic high fat diet on GLP-1 sensitivity in vagal afferents. C57/BL6 mice were fed either a high-fat or low-fat diet for 6–8 weeks. To evaluate gastrointestinal afferent sensitivity and nodose neurons’ response to GLP-1, extracellular afferent recordings and patch clamp were performed, respectively. Exendin-4 (Ex-4) was used as an agonist of the GLP-1 receptor. C-Fos Expression was examined as an indication of afferent input to the nucleus tractus solitarius (NTS). Food intake was monitored in real-time before and after Ex-4 treatment to monitor the consequence of the high fat diet on the satiating effect of GLP-1. In high fat fed (HFF) mice, GLP-1 caused lower activation of intestinal afferent nerves, and failed to potentiate mechanosensitive nerve responses compared to low fat fed (LFF). GLP-1 increased excitability in LFF and this effect was reduced in HFF neurons. Consistent with these findings on vagal afferent nerves, GLP-1 receptor stimulation given systemically, had a reduced satiating effect in HFF compared to LFF mice, and neuronal activation in the NTS was also reduced. The present study demonstrated chronic high fat diet impaired vagal afferent responses to GLP-1, resulting in impaired satiety signaling. GLP-1 sensitivity may account for the impairment of satiety signaling in obesity and thus a therapeutic target for obesity treatment.

Cholera and Pancreatic Cholera: Is VIP the Common Pathophysiologic Factor?

Background: Cholera remains a major global health problem, causing high output diarrhea leading to severe dehydration and shock in developing countries. We aimed to determine whether vasoactive intestinal polypeptide (VIP), the mediator of pancreatic cholera syndrome, has a role in the pathophysiology of human cholera. Methods: We conducted a prospective observational study of cholera cases hospitalized with severe dehydration. Plasma and stool water levels of VIP were measured just after admission, after complete rehydration (3–4 h), at 24 h post-rehydration and at discharge after diarrhea ceased. Results: In total, 23 cholera patients were examined between January and August 2018. The geometric mean of stool VIP (sVIP) and plasma VIP (pVIP) on admission were 207.67 and 8.34 pmol/L, respectively. pVIP values were all within the normal range (</= 30 pcmol/L); however, sVIP levels were very high at all timepoints, though less so just after rehydration. In multivariable GEE models, after adjustment for covariates, sVIP levels were significantly associated with duration of hospitalization (p = 0.026), total stool volume (p = 0.023) as well as stool output in the first 24 h (p = 0.013). Conclusions: The data suggest that VIP, which is released by intestinal nerves, may play an important role in human choleragenesis, and inhibitors of intestinal VIP merit testing for potential therapeutic benefits.

Cannabis Hyperemesis Syndrome: A Still Under-Recognized Syndrome.

Cannabis is one of the most widely used illicit drugs in the world. Its use is associated with several short- and long-term side-effects such as changes in mood, impaired memory, impaired attention, depression and anxiety, and it is correlated with schizophrenia. Cannabinoid hyperemesis syndrome (CHS) is characterized by chronic cannabis use, cyclic intractable nausea and vomiting, and compulsive hot bathing. Patients are typically diagnosed with CHS only after multiple medical evaluations. Recent research has identified type 1 cannabinoid receptors in the intestinal nerve plexus that have an inhibitory effect on gastrointestinal motility. This effect may explain hyperemesis in cannabis users. The thermoregulatory role of endocannabinoids may be responsible for compulsive hot bathing. We describe the cases of two young men seeking repeated emergency room care with recurrent nausea and vomiting. Abstinence from cannabis led to resolution of vomiting symptoms and abdominal pain.LEARNING POINTSCannabis is one of the most widely used illicit drugs in world.Cannabinoid hyperemesis syndrome is an under-recognized syndrome that can lead to expensive and unrevealing investigations.Abstinence from cannabis led to resolution of symptoms.

Spatial relationship between telocytes, interstitial cells of Cajal and the enteric nervous system in the human ileum and colon.

Telocytes (TCs) are recently described interstitial cells, present in almost all human organs. Among many other functions, TCs regulate gastrointestinal motility together with the interstitial cells of Cajal (ICCs). TCs and ICCs have close localization in the human myenteric plexus; however, the exact spatial relationship cannot be clearly examined by previously applied double immunofluorescence/confocal microscopy. Data on TCs and submucosal ganglia and their relationship to intestinal nerves are scarce. The aim of the study was to analyse the spatial relationship among these components in the normal human ileum and colon with double CD34/CD117 and CD34/S100 immunohistochemistry and high‐resolution light microscopy. TCs were found to almost completely encompass both myenteric and submucosal ganglia in ileum and colon. An incomplete monolayer of ICCs was localized between the TCs and the longitudinal muscle cells in ileum, whereas only scattered ICCs were present on both surfaces of the colonic myenteric ganglia. TC‐telopodes were observed within colonic myenteric ganglia. TCs, but no ICCs, were present within and around the interganglionic nerve fascicles, submucosal nerves and mesenterial nerves, but were only observed along small nerves intramuscularly. These anatomic differences probably reflect the various roles of TCs and ICCs in the bowel function.

Induced pluripotent stem cell-derived enteric neural crest cells repopulate human aganglionic tissue-engineered intestine to form key components of the enteric nervous system

Models for enteric neuropathies, in which intestinal nerves are absent or injured, are required to evaluate possible cell therapies. However, existing options, including transgenic mice, are variable and fragile. Here immunocompromised mice were implanted with human pluripotent stem cell–derived tissue-engineered small intestine 10 weeks prior to a second survival surgery in which enteric nervous system precursor cells, or saline controls, were injected into the human intestinal organoid–derived tissue-engineered small intestine and analyzed 4 weeks later. Human intestinal organoid–derived tissue-engineered small intestine implants injected with saline as controls illustrated formation of intestinal epithelium and mesenchyme without an enteric nervous system. Second surgical introduction of human pluripotent stem cell–generated enteric nervous system precursors into developing human intestinal organoid–derived tissue-engineered small intestine implants resulted in proliferative migratory neuronal and glial cells, including multiple neuronal subtypes, and demonstrated function in contractility assays.