Projections of peptide-containing neurons in rat colon

doi:10.1016/0306-4522(88)90296-5

Neuroscience

Volume 27, Issue 2, November 1988, Pages 655-674

https://doi.org/10.1016/0306-4522(88)90296-5 Get rights and content

Abstract

The distribution, origin and projections of nerve fibers containing vasoactive intestinal peptide, substance P, neuropeptide Y, galanin, gastrin-releasing peptide, calcitonin gene-related peptide, somatostatin or enkephalin were studied in the midcolon of the rat by immunocytochemistry and immunochemistry. Most of these nerve fibers had an intramural origin as was established by extrinsic denervation (severing of mesenterial nerves). Extrinsic denervation eliminated neuropeptide Y-contaming fibers of presumably sympathetic origin together with sensory nerve fibers containing both substance P and calcitonin gene-related peptide. Co-existence of two peptides in the same neuron was studied by double immunostaining. This revealed co-existence of neuropeptide Y and vasoactive intestinal peptide in one population of intramural neurons; an additional population of intramural neurons was found to contain vasoactive intestinal peptide but not neuropeptide Y. All somatostatin-containing neurons in the submucous ganglia were found to harbor calcitonin gene-related peptide. A much larger population of submucous neurons containing calcitonin gene-related but not somatostatin was also detected. Some perivascular calcitonin gene-related peptide-containing nerve fibers (of intrinsic origin) harbored vasoactive intestinal peptide while others (of extrinsic origin) harbored substance P. The polarities and projections of the various peptide-containing intramural neurons in the transverse colon were studied by analysing the loss of nerve fibers upon local disruption of enteric nervous pathways (myectomy or intestinal clamping). Myenteric neurons containing vasoactive intestinal peptide, galanin, gastrin-releasing peptide, calcitonin gene-related peptide, somatostatin or vasoactive intestinal peptide/neuropeptide Y gave off 5–10-mm-long descending projections while those containing substance P or enkephalin issued approx. 5-mm-long ascending projections. Submucous neurons containing calcitonin gene-related peptide, somatostatin/calcitonin gene-related peptide or gastrin-releasing peptide issued both ascending (2–6 mm) and descending (2–6 mm) projections, those containing vasoactive intestinal peptide issued ascending (approx. 2 mm) projections, while those containing galanin or vasoactive intestinal peptide/neuropeptide Y lacked demonstrable oro-anal projections. Enkephalin-containing fibers could not be detected in the mucosa and the mucosal substance P-containing nerve fibers were too few to enable us to delineate their projections.

Reference (36)

EkbladE. et al.
VIP and PHI coexist with an NPY-like peptide in intramural neurones of the small intestine
Regul. Pept.
(1984)
EkbladE. et al.
Peptide-containing nerve fibers in the stomach wall of rat and mouse
Gastroenterology
(1985)
EkbladE. et al.
Galanin nerve fibers in the rat gut: distribution, origin and projections
Neuroscience
(1985)
EkbladE. et al.
Projections of peptide-containing neurons in rat small intestine
Neuroscience
(1987)
FurnessJ.B. et al.
Polarity and projections of peptide-containing neurons in the guinea pig small intestine
FurnessJ.B. et al.
Distribution and projections of nerves with enkephalin-like immunoreactivity in the guinea-pig small intestine
Neuroscience
(1983)
GibbinsI.L. et al.
Co-localization of calcitonin gene-related peptide-like immunoreactivity with substance P in cutaneous, vascular and visceral sensory neurons of guinea pigs
Neurosci. Lett.
(1985)
GrunditzT. et al.
Neuronal pathways to the rat thyroid revealed by retrograde tracing and immunocytochemistry
Neuroscience
(1988)
KeastJ.R. et al.
Origins of peptide and norepinephrine nerves in the mucosa of the guinea pig small intestine
Gastroenterology
(1984)
MakhloufG.M.
Enteric neuropeptides: role in neuromuscular activity of the gut
Trends pharmac. Sci.
(1985)

MoghimzadehE. et al.

Neuronal gastrin-releasing peptide in the mammalian gut and pancreas

Neuroscience

(1983)

SundlerF. et al.

Peptidergic nervous systems in the gut

Clin. Gastroenterol.

(1980)

AlumetsJ. et al.

Leu-enkephalin-like material in nerves and enterochromaffin cells in the gut

Histochemistry

(1978)

BrodinE. et al.

Immunoreactive substance P in the chicken gut: distribution, development and possible functional significance

Cell Tiss. Res.

(1981)

BrodinE. et al.

Tachykinins in rat central nervous system: distribution, molecular forms, release and effects of chronic treatment with antidepressant drugs

ClaqueJ.R. et al.

Localization of calcitonin gene-related peptide-like immunoreactivity in neurons of the rat gastrointestinal tract

Neurosci. Lett.

(1985)

CoonsA.H. et al.

Studies on antibody production I. A method for the histochemical demonstration of specific antibody and its application to a study of the hyperimmune rabbit

J. exp. Med.

(1955)

CostaM. et al.

Nervous control of intestinal motility

Cited by (119)

Peripheral Autonomic Nervous System
2015, The Rat Nervous System: Fourth Edition
The autonomic nervous system adjusts the activities of organs and tissues that are not under overt voluntary control so that they operate at levels most favorable to the states of the body and environment. It has parasympathetic, sympathetic and enteric divisions, plus associated afferent systems that detect body and environmental conditions relevant to autonomic control. Parasympathetic motor pathways leave the central nervous system in the 3rd, 7th, 9th and 10th cranial nerves and in sacral nerves. Sympathetic pathways emerge from the thoracic and lumbar spinal cord. Autonomic pathways form synapses in cranial, cervical, thoracic, abdominal and pelvic ganglia where signals are integrated and relayed. Many tissues are innervated only by parasympathetic or sympathetic neurons. Some organs are under dual, complementary, control. Ganglia of the enteric division are in the walls of the digestive tract, gallbladder and pancreas. In the intestine the enteric division includes full reflex pathways (afferent neurons, interneurons and motor neurons).
Enteric Nervous System Structure and Neurochemistry Related to Function and Neuropathology
2012, Physiology of the Gastrointestinal Tract, Two Volume Set
Enteric Nervous System Structure and Neurochemistry Related to Function and Neuropathology
2012, Physiology of the Gastrointestinal Tract
Mast cells reduce survival of myenteric neurons in culture
2009, Neuropharmacology
Citation Excerpt :
As general neuronal markers' antibodies against HuC/HuD (a mouse monoclonal anti-human neuronal protein; code no. A-21271; Molecular Probes, Eugene, OR, USA; dilution 1:400; Lin et al., 2003) or human protein gene product 9.5 (code no. PGP 9.5; Ultraclone, Isle of Wight, UK; dilution 1:1600; Sandgren et al., 2003) were used. For the detection of VIP- and NOS-containing neurons a polyclonal VIP antiserum against porcine VIP raised in rabbit (code no. 7852; Euro-Diagnostica AB, Malmö, Sweden; dilution 1:1200; Ekblad et al., 1988) or a polyclonal antiserum against synthetic rat cerebellar NOS (code no. 9223; Euro-Diagnostica AB, Malmö, Sweden; dilution 1:5000; Ekblad et al., 1994) were used in double-staining procedures with anti-Hu. Cultured mast cells were detected by antibodies raised in rabbits against histamine (code no. 8432, Euro-Diagnostica AB, Malmö, Sweden; dilution 1:300, Håkanson et al., 1986).
Mast cell–nerve interactions play a key role in intestinal inflammation and irritable bowel disease. Loss of enteric neurons has been reported in inflammatory conditions but the contribution of mast cells in this event is unknown. To study neuronal survival and plasticity of myenteric neurons in contact with mast cells a co-culture system using myenteric neurons from rat small intestine and peritoneal mast cells was set up.
Dissociated myenteric neurons were cultured for 4 days before addition of mast cells isolated by peritoneal lavage. Neuronal survival and expression of vasoactive intestinal peptide (VIP) and nitric oxide synthase (NOS) were studied by immunocytochemistry and neuronal cell counting. Myenteric neurons cultured without mast cells were used to study the rate of neuronal survival after the addition of various mast cell mediators, proteinase-activated receptor₂ (PAR₂) agonist, VIP or corticosteroid.
A striking mast cell-induced neuronal cell death was found after co-culturing. It was counteracted by the addition of mast cell stabiliser doxantrazole, protease inhibitors, PAR₂ antagonist FSLLRY-amide, corticosteroid or VIP. In myenteric neurons cultured without mast cells the PAR₂ agonist SLIGRL-amide, prostaglandin D₂ and interleukin (IL) 6 reduced neuronal survival while histamine, serotonin, heparin, IL1β and tumour necrosis factor α had no effect; corticosteroid and VIP enhanced neuronal survival. The relative numbers of VIP-, but not NOS-expressing myenteric neurons increased after co-culturing.
Mast cell-induced neuronal cell death is suggested to be mediated via PAR₂ activation, IL6 and prostaglandin D₂. Corticosteroid and VIP are neuroprotective and able to prevent cell death of myenteric neurons in co-culture.
Colonic Vascular Conductance Increased by Daikenchuto via Calcitonin Gene-Related Peptide and Receptor-Activity Modifying Protein 1
2008, Journal of Surgical Research
Citation Excerpt :
Nervous mechanisms are important for the regulation of gastrointestinal blood flow. A number of neuropeptides such as CGRP, VIP, and SP have been localized immunohistochemically in sensory nerves innervating various viscera, including the gastrointestinal tract [29–33]. Exogenous application of these peptides has been shown to dilate arterioles [34–36].
Daikencyuto (DKT) is a traditional Japanese medicine (Kampo) and is a mixture of extract powders from dried Japanese pepper, processed ginger, ginseng radix, and maltose powder and has been used as the treatment of paralytic ileus. DKT may increase gastrointestinal motility by an up-regulation of the calcitonin gene-related peptide (CGRP). CGRP is also the most powerful vasoactive substance. In the present study, we investigated whether DKT has any effect on the colonic blood flow in rats.
Experiments were performed on fasted anesthetized and artificially ventilated Wistar rats. Systemic mean arterial blood pressure and heart rate were recorded. Red blood cell flux in colonic blood flow was measured using noncontact laser tissue blood flowmetry, and colonic vascular conductance (CVC) was calculated as the ratio of flux to mean arterial blood pressure. We examined four key physiological mechanisms underlying the response using blocker drugs: CGRP1 receptor blocker (CGRP_8–37), nitric oxide synthase inhibitor, vasoactive intestinal polypeptide (VIP) receptor blocker ([4-Cl-DPhe6, Leu17]-VIP), and substance P receptor blocker (spantide). Reverse transcription-polymerase chain reaction was used for the detection of mRNA of calcitonin receptor-like receptor, receptor-activity modifying protein 1, the component of CGRP 1 receptor and CGRP. After laparotomy, a cannula was inserted into the proximal colon to administer the DKT and to measure CVC at the distal colon.
Intracolonal administration of DKT (10, 100, and 300 mg/kg) increased CVC (basal CVC, 0.10 mL/mmHg) from the first 15-min observation period (0.14, 0.17, and 0.17 mL/mmHg, respectively) and with peak response at either 45 min (0.17 mL/mmHg by 10 mg/kg), or 75 and 60 min (0.23 and 0.21 mL/mmHg by 100 and 300 mg/kg, respectively). CGRP_8–37 completely abolished the DKT-induced hyperemia, whereas nitric oxide synthase inhibitor partially attenuated the DKT-induced hyperemia. [4-Cl-DPhe6, Leu17]-VIP and spantide did not affect the hyperemia. Japanese pepper significantly increased CVC at 45 min or later, whereas ginseng radix only showed a significant increase at 15 min. Reverse transcription-polymerase chain reaction showed that mRNA for calcitonin receptor-like receptor, receptor-activity modifying protein 1, and CGRP were expressed in rat colon and up-regulated by DKT.
The present study demonstrated that DKT increased CVC, which was mainly mediated by CGRP and its receptor components.
Endogenous PYY and NPY mediate tonic Y<inf>1</inf>- and Y<inf>2</inf>-mediated absorption in human and mouse colon
2008, Nutrition
Citation Excerpt :
In the enteric nervous system, NPY is present in ∼50% of submucous plexus neurons, the majority of which innervate the mucosa and target the lamina propria of most species, including rat, mouse, and human colon [12,13]. Within the submucous plexus, NPY is most frequently present in secretomotor nerves and is colocalized with the inhibitory neurotransmitter, vasoactive intestinal polypeptide [12,14]. PP, in contrast, is present in a sparse population of endocrine cells scattered along the length of the intestine in most species [9,10], its primary source being the pancreatic F-cells.
To establish the functional significance of endogenous peptide YY (PYY) and neuropeptide Y (NPY) as mediators of Y₁ and Y₂ absorptive tone in colonic mucosa.
Functional studies utilized descending colon from adult mice (wild type [WT] and peptide nulls) and ex vivo human colonic tissue (from patients undergoing bowel resections) measuring changes in basal ion transport. Peak increases in ion transport to Y₁ or Y₂ antagonists (BIBO3304 300 nM; BIIE0246 1 μM) were pooled (mean ± SEM) and compared using Student's unpaired t test (P ≤ 0.05); some tissues received tetrodotoxin (TTX; 100 nM). PYY-positive L-cell numbers and NPY innervation were also compared.
Y₁ and Y₂ tones were present in human and WT mouse colon mucosa and only the latter was TTX sensitive. Y₁ tone was unchanged in NPY^−/− but was ∼90% inhibited in PYY^−/− and abolished in PYYNPY^−/− colon mucosa. Y₂ tone was reduced ∼50% in NPY^−/− and PYY^−/− tissues and was absent from PYYNPY^−/− colon. Residual Y₂ and Y₁ tones present in PYY^−/− mucosa were abolished by TTX. PYY ablation had no apparent effect on NPY innervation and PYY-positive cells were observed at the same frequency in NPY^−/− (56.7 ± 6.8 cells/section) and WT (55.0 ± 4.6 cells/section) colons. Double knockouts lacked PYY and NPY expression, but endocrine cells and enteric nerves were present with similar frequencies to those of WT mice.
Endogenous PYY mediates Y₁ absorptive tone that is epithelial in origin, whereas Y₂ tone is a combination of PYY and NPY mediation.

View all citing articles on Scopus

View full text

Projections of peptide-containing neurons in rat colon

Abstract

Regul. Pept.

Gastroenterology

Neuroscience

Neuroscience

Neuroscience

Neurosci. Lett.

Neuroscience

Gastroenterology

Trends pharmac. Sci.

Neuroscience

Clin. Gastroenterol.

Leu-enkephalin-like material in nerves and enterochromaffin cells in the gut

Histochemistry

Immunoreactive substance P in the chicken gut: distribution, development and possible functional significance

Cell Tiss. Res.

Tachykinins in rat central nervous system: distribution, molecular forms, release and effects of chronic treatment with antidepressant drugs

Localization of calcitonin gene-related peptide-like immunoreactivity in neurons of the rat gastrointestinal tract

Neurosci. Lett.

Studies on antibody production I. A method for the histochemical demonstration of specific antibody and its application to a study of the hyperimmune rabbit

J. exp. Med.

Nervous control of intestinal motility