Myenteric neurons and interstitial cells of Cajal of mouse colon express several nitric oxide synthase isoforms

doi:10.1016/S0304-3940(02)00338-5

Neuroscience Letters

Volume 326, Issue 3, 5 July 2002, Pages 191-195

https://doi.org/10.1016/S0304-3940(02)00338-5 Get rights and content

Abstract

Information on equipment and subcellular distribution of nitric oxide synthase (NOS) isoforms in myenteric neurons and pacemaker cells (ICC) might help to identify nitric oxide (NO) pathway(s) acting on gastrointestinal motility. In sections of mouse colon labelled with neuronal (n)NOS, endothelial (e)NOS and inducible (i)NOS antibodies, all myenteric neurons co-expressed eNOS and iNOS and a subpopulation of them co-expressed nNOS. ICC co-expressed nNOS and eNOS. In the neurons, nNOS-labeling was intracytoplasmatic, in the ICC at cell periphery. In both cell types, eNOS-labeling was on intracytoplasmatic granules, likely mitochondria. In conclusion, myenteric neurons and ICC co-express several NOS isoforms with specific subcellular distribution. Different nNOS splice variants are presumably present: intracytoplasmatic nNOSβ and nNOSα producing neurogenic NO, plasma membrane-bound nNOSα producing ICCgenic NO. eNOS might be implicated in mitochondrial respiration and, in ICC, also in pacemaker activity. Neurons express iNOS also in basal condition.

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Acknowledgements

The financial support of Telethon-Italy (Grant no 1134 to M-S F-P) is gratefully acknowledged.

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A 22-year-old male patient having Duchenne muscular dystrophy (DMD) and chronic constipation referred to the emergency department of our hospital for severe abdominal pain and signs of bowel obstruction. Contrast-enhanced computed tomography of the abdomen demonstrated mechanical ileus due to a volvulus of the transverse colon. Torsion of the transverse mesocolon and multiple ischemic areas with focal intestinal wall perforations were confirmed during surgical exploration and a subtotal colectomy was performed. DMD is frequently associated to gastrointestinal motility disorders, including chronic constipation, and life-threatening conditions like intestinal pseudo-obstruction and sigmoid volvulus. To date to our knowledge, transverse colon volvulus represents an unreported condition among patients with DMD.
Spontaneous transverse colon volvulus in a patient with Duchenne muscular dystrophy: An unreported complication
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A 22-year-old male patient with Duchenne muscular dystrophy (DMD) and chronic constipation presents to the emergency room with severe abdominal pain and hive closed to feces and gas. Contrast-enhanced computed tomography of the abdomen demonstrates mechanical ileus due to volvulus of the transverse colon: torsion of the transverse mesocolon is confirmed and subtotaly colectomy is performed, revealing multiple ischemic areas with focal perforations. DMD is frequently associated with gastrointestinal motility disorders, including chronic constipation and life-threatening conditions like intestinal pseudo-obstruction and sigmoid volvulus. To date, transverse colic localization of volvolus represents an unreported condition among patients with DMD.
Increase of n-NOS and i-NOS in Rat Colon After Sacral Neuromodulation
2017, Neuromodulation
Citation Excerpt :
Wang et al. reported that NO could be produced in the ENS through both n-NOS and i-NOS pathway and ENS might be involved in the inhibition of the contraction of the circular muscle strips (13). Active i-NOS gene is present in the myenteric neurons in basal conditions (36), so the amount and activity of i-NOS might be more tightly controlled than in other kinds of cells. Jing et al. showed that NO produced by i-NOS played a predominant effect on the contraction of circular muscle strips from colon in male rats (31).
Sacral neuromodulation (SNM) is proposed to treat different anorectal dysfunctions but its mechanism of action is not yet known. Our previous study demonstrated how SNM can significantly increase neuronal nitric oxide synthase NOS (n-NOS) and inducible NOS (i-NOS) expression in the anus and rectum of rats. There are no reports regarding the relation between SNM and NOS in colonic cells: our aim was to assess NOS expression in colonic rat model after SNM.
Twenty-six female Sprangue-Dawley rats were considered: group I, normal control rats; group II, sham treatment rats, in whom electrodes for electrical stimulation were placed in S1 foramen bilaterally and left in place, without performing neuromodulation; group III, rats in whom SNM was performed. After 14 days, the rats were sacrificed and we evaluated n-NOS and i-NOS in colonic specimens by immunohistochemistry and Western Blot analysis.
Western Blot analysis showed that levels of n-NOS and i-NOS were higher in colon of the III group rats respect to the others; in particular, immunohistochemistry revealed that, after neuromodulation, n-NOS expression in the muscle cells and i-NOS expression in glandular epithelium and nervous cells were highly represented (p < 0.05).
Our study showed that in colon, SNM is able to influence NO synthesis, activating n-NOS expression in muscle cells and i-NOS expression in glandular epithelium and nervous cells. Our study showed a complex colonic response to SNM. This experimental model could be applied to better understand the mechanism of action of SNM in bowel dysfunction.
Inhibiting inducible nitric oxide synthase in enteric glia restores electrogenic ion transport in mice with colitis
2015, Gastroenterology
Disturbances in the control of ion transport lead to epithelial barrier dysfunction in patients with colitis. Enteric glia regulate intestinal barrier function and colonic ion transport. However, it is not clear whether enteric glia are involved in epithelial hyporesponsiveness. We investigated enteric glial regulation of ion transport in mice with trinitrobenzene sulfonic acid− or dextran sodium sulfate−induced colitis and in Il10^−/− mice.
Electrically evoked ion transport was measured in full-thickness segments of colon from CD1 and Il10^−/− mice with or without colitis in Ussing chambers. Nitric oxide (NO) production was assessed using amperometry. Bacterial translocation was investigated in the liver, spleen, and blood of mice.
Electrical stimulation of the colon evoked a tetrodotoxin-sensitive chloride secretion. In mice with colitis, ion transport almost completely disappeared. Inhibiting inducible NO synthase (NOS2), but not neuronal NOS (NOS1), partially restored the evoked secretory response. Blocking glial function with fluoroacetate, which is not a NOS2 inhibitor, also partially restored ion transport. Combined NOS2 inhibition and fluoroacetate administration fully restored secretion. Epithelial responsiveness to vasoactive intestinal peptide was increased after enteric glial function was blocked in mice with colitis. In colons of mice without colitis, NO was produced in the myenteric plexus almost completely via NOS1. NO production was increased in mice with colitis, compared with mice without colitis; a substantial proportion of NOS2 was blocked by fluoroacetate administration. Inhibition of enteric glial function in vivo reduced the severity of trinitrobenzene sulfonic acid−induced colitis and associated bacterial translocation.
Increased production of NOS2 in enteric glia contributes to the dysregulation of intestinal ion transport in mice with colitis. Blocking enteric glial function in these mice restores epithelial barrier function and reduces bacterial translocation.
Interaction between NMDA glutamatergic and nitrergic enteric pathways during in vitro ischemia and reperfusion
2015, European Journal of Pharmacology
Nitric oxide (NO) and glutamate, via N-methyl-d-aspartate (NMDA) receptors, participate to changes in neuromuscular responses after ischemic/reperfusion (I/R) injury in the gut. In the present study we investigated the existence of a possible interplay between nitrergic and NMDA receptor pathways in the guinea pig ileum after in vitro I/R injury, resorting to functional and biomolecular approaches. In normal metabolic conditions NMDA concentration-dependently enhanced both glutamate (analyzed by high performance liquid chromatography with fluorimetric detection) and NO (spectrophotometrically quantified as NO₂⁻ and NO₃⁻) spontaneous overflow from isolated ileal segments. Both effects were reduced by the NMDA antagonists, (-)-AP5 (10 µM) and 5,7-diCl-kynurenic acid (10 µM, 5,7-diCl-KYN). N^ω-propyl-l-arginine (1 µM, NPLA) and 1400W (10 µM), respectively, nNOS and iNOS inhibitors, reduced NMDA-stimulated glutamate overflow. After in vitro I/R, glutamate overflow increased, and returned to control values in the presence of NPLA and 1400W. NO₂⁻ and NO₃⁻ levels transiently increased during I/R and were reduced by both (-)-AP5 and 5,7-diCl-KYN. In longitudinal muscle myenteric plexus preparations, iNOS mRNA and protein levels increased after in vitro I/R; both parameters were reduced to control values by (-)-AP5 and 5,7-diCl-KYN. Both antagonists were also able to reduce ischemia-induced enhancement of nNOS mRNA levels. Protein levels of GluN1, the ubiquitary subunit of NMDA receptors, increased after I/R and were reduced by both NPLA and 1400W. On the whole, this data suggests the existence of a cross-talk between NMDA receptor and nitrergic pathways in guinea pig ileum myenteric plexus, which may participate to neuronal rearrangements occurring during I/R.
Effects of menthol on circular smooth muscle of human colon: Analysis of the mechanism of action
2014, European Journal of Pharmacology
Citation Excerpt :
Because antispasmodic activity of peppermint oil in rat trachea involves NO-synthase (de Sousa et al., 2010), the present study verified the putative contribute of NO in the inhibitory responses to menthol. On the other hand, NO is one of the main inhibitory neurotransmitters in the gastrointestinal tract, including human colon (Gallego et al., 2008) and can originate from non-neural cells of the intestinal tract, such as smooth muscle cells (Murthy and Makhlouf, 1994; Xue et al., 1994; Jin et al., 1996; Teng et al., 1998) or interstitial cells of Cajal (Vannucchi et al., 2002; Fornai et al., 2007). NO signalling involves activation of cytosolic guanylyl cyclase with elevation of cGMP, activation of protein kinase G and consequent cellular hyperpolarization, likely due K+ channel activation.
Menthol is the major constituent of peppermint oil, an herbal preparation commonly used to treat nausea, spasms during colonoscopy and irritable bowel disease. The mechanism responsible for its spasmolytic action remains unclear. The aims of this study were to investigate the effects induced by menthol on the human distal colon mechanical activity in vitro and to analyze the mechanism of action. The spontaneous or evoked-contractions of the circular smooth muscle were recorded using vertical organ bath. Menthol (0.1 mM–30 mM) reduced, in a concentration-dependent manner, the amplitude of the spontaneous contractions without affecting the frequency and the resting basal tone. The inhibitory effect was not affected by 5-benzyloxytryptamine (1 μM), a transient receptor potential-melastatin8 channel antagonist, or tetrodotoxin (1 μM), a neural blocker, or 1H-[1,2,4] oxadiazolo [4,3-a]quinoxalin-1-one (10 µM), inhibitor of nitric oxide (NO)-sensitive soluble guanylyl cyclase, or tetraethylammonium (10 mM), a blocker of potassium (K⁺)-channels. On the contrary, nifedipine (3 nM), a voltage-activated L-type Ca²⁺ channel blocker, significantly reduced the inhibitory menthol actions. Menthol also reduced in a concentration-dependent manner the contractile responses caused by exogenous application of Ca²⁺ (75–375 μM) in a Ca²⁺-free solution, or induced by potassium chloride (KCl; 40 mM). Moreover menthol (1–3 mM) strongly reduced the electrical field stimulation (EFS)-evoked atropine-sensitive contractions and the carbachol-contractile responses. The present results suggest that menthol induces spasmolytic effects in human colon circular muscle inhibiting directly the gastrointestinal smooth muscle contractility, through the block of Ca²⁺ influx through sarcolemma L-type Ca²⁺ channels.

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Myenteric neurons and interstitial cells of Cajal of mouse colon express several nitric oxide synthase isoforms

Abstract

Section snippets

Acknowledgements

FEBS Lett.

Neurosci. Lett.

Neurosci. Lett.

Gastroenterology

Neurosci. Lett.

J. Auton. Nerv. Syst.

Nitric oxide from enteric nerves acts by a different mechanism from myogenic nitric oxide in canine lower esophageal sphincter

J. Pharmacol. Exp.Ther.

Endothelial nitric oxide synthase localized to hippocampal pyramidal cells: implications for synaptic plasticity

Proc. Natl. Acad. Sci. USA

Subcellular localization and characterization of neuronal nitric oxide synthase (NOS)

J. Neurochem.

Characterization and splice variants of neuronal nitric oxide synthase in rat small intestine

Am. J. Physiol.