Neurochemically similar myenteric and submucous neurons directly traced to the mucosa of the small intestine

Furness, John B.; Costa, M.; Gibbins, Ian L.; Llewellyn‐Smith, Ida J.; Oliver, James M.

doi:10.1007/bf00214637

Cited by 185 publications

(79 citation statements)

References 19 publications

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“…This is supported by the abolition of staining by preadsorption of 1.B3.9B3 with human placental ChAT and the lack of staining with an irrelevant, isotypematched monoclonal antibody (anti-hepatitis B surface antigen). The localization of ChAT IR in the submucosal and myenteric plexi is consistent with other reports that employed immunocytochemistry on whole mounts (Furness et al 1984(Furness et al ,1985Schemann et al 1993Schemann et al ,1995Mann et al 1995), including studies that successfully used 1.B3.9B3 (Schemann et al 1993(Schemann et al ,1995. In addition, ChAT IR and activity in endothelia of various tissues have been previously described (Parnavelas et al 1985;Gonzalez and Santos-Benito 1987;Milner et al 1989), and there are reports of ChAT in epithelial cells (Grando et al 1993;Sakuragawa et al 1997) and lymphocytes (Rinner and Schauenstein 1993).…”

Section: Discussionsupporting

confidence: 88%

“…Reports using AChE histochemistry on normal mucosa either have dubious specificity (Isaacs et al 1976) or report few or no cholinergic nerve fibers (Lake et al 1978). Studies with ChAT immunohistochemistry on whole mounts have traced ChAT IR nerve processes to the mucosa but have not demonstrated ChAT IR nerve fibers in this compartment (Furness et al 1985). Possibly, ChAT is not transported into mucosal nerve processes, or a non-1.B3.9B3-immunoreactive ChAT isoenzyme is present in mucosal nerves (for a discussion of ChAT expression, see Mallet et al 1990).…”

Section: Figurementioning

confidence: 99%

“…However, this enzyme is found in relatively small amounts in neural tissue, complicating the development of immunostaining methods in the enteric nervous system (Schemann et al 1993). Therefore, immunocytochemistry for ChAT has as yet been applied only to frozen sections and whole mounts in a limited number of studies in the enteric nervous system (Furness et al 1984(Furness et al ,1985Schemann et al 1993Schemann et al ,1995Mann et al 1995). Whole mounts and frozen section preparations do not permit the same microscopic resolution as paraffin sections and preclude the use of archival tissue.…”

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confidence: 99%

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Choline Acetyltransferase (ChAT) Immunoreactivity in Paraffin Sections of Normal and Diseased Intestines

Ratcliffe

Desa

Dixon

et al. 1998

J Histochem Cytochem.

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SUMMARYThere is increasing interest in localizing nerves in the intestine, especially specific populations of nerves. At present, the usual histochemical marker for cholinergic nerves in tissue sections is acetylcholinesterase activity. However, such techniques are applicable only to frozen sections and have uncertain specificity. Choline acetyltransferase (ChAT) is also present in cholinergic nerves, and we therefore aimed to establish a paraffin section immunocytochemical technique using an anti-ChAT antibody. Monoclonal anti-choline acetyltransferase (1.B3.9B3) and a biotin-streptavidin detection system were used to study the distribution of ChAT immunoreactivity (ChAT IR) in paraffin-embedded normal and diseased gastrointestinal tracts from both rats and humans. Optimal staining was seen after 6-24 hr of fixation in neutral buffered formalin and overnight incubation in 1 g/ml of 1.B3.9B3, with a similar distribution to that seen in frozen sections. In the rat diaphragm (used as a positive control), axons and motor endplates were ChAT IR. Proportions of ganglion cells and nerve fibers in the intramural plexi of both human and rat gastrointestinal tracts were also ChAT IR, as well as extrinsic nerve bundles in aganglionic segments of Hirschsprung's disease. Mucosal cholinergic nerves, however, were not visualized. In addition, non-neuronal cells such as endothelium, epithelium, and inflammatory cells were ChAT IR. We were able to localize ChAT to nerves in formalin-fixed, paraffin-embedded sections. The presence of ChAT IR in non-neuronal cells indicates that this method should be used in conjunction with other antibodies. Nevertheless, it proves to be a useful technique for studying cholinergic neuronal distinction in normal tissues and pathological disorders.

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Section: Discussionsupporting

confidence: 88%

Section: Figurementioning

confidence: 99%

mentioning

confidence: 99%

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Choline Acetyltransferase (ChAT) Immunoreactivity in Paraffin Sections of Normal and Diseased Intestines

Ratcliffe

Desa

Dixon

et al. 1998

J Histochem Cytochem.

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“…Wholemounts, fixed and prepared as above, were incubated in primary antisera against NPY (sheep anti-NPY, E2210, 1:400 (Furness et al, 1985)). After the preparations were washed they were incubated with biotinylated donkey anti-sheep IgG, 1:200.…”

Section: Methodsmentioning

confidence: 99%

“…These four types include 1) neurons with immunoreactivity for vasoactive intestinal peptide (VIP) and several other peptides, including galanin, dynorphin, and neuromedin U; 2) neurons immunoreactive for neuropeptide Y (NPY), choline acetyltransferase (ChAT), somatostatin, and calcitonin gene-related peptide; 3) neurons immunoreactive for calretinin, ChAT, and dynorphin; and 4) neurons with a typical Dogiel type II morphology that are immunoreactive for ChAT and tachykinins, about half of which are also immunoreactive for calbindin. NPY immunoreactivity defines the shapes of one phenotype (Furness et al, 1985), but neither VIP nor calretinin, nor other markers of the VIP and calretinin phenotypes, reveal the dendritic morphologies of these types (Furness et al, 1984;Brookes et al, 1991).…”

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confidence: 96%

Morphologies and projections of defined classes of neurons in the submucosa of the guinea‐pig small intestine

et al. 2003

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Four types of neurons have previously been identified by neurochemical markers in the submucosal ganglia of the guinea-pig small intestine, and functional roles have been ascribed to each type. However, morphological differences among the classes have not been determined, and there is only partial information about their projections within the submucosa. In the present work, we used intracellular microelectrodes to fill neurons of each type with biocytin, which was then converted to a permanent dye, so that the shapes of the neurons could be determined and their projections within the submucosa could be followed. Cell bodies of noncholinergic secretomotor/ vasodilator neurons had Dogiel type I morphology. These neurons, which are vasoactive intestinal peptide immunoreactive, had single axons that ran through many ganglia without providing terminals around other neurons. Cholinergic secretomotor neurons with neuropeptide Y immunoreactivity had Stach type IV morphology, and cholinergic secretomotor/vasodilator neurons had stellate cell bodies. The axons of these two types ran short distances in the plexus and did not innervate other submucosal neurons. Neurons of the fourth type, intrinsic primary afferent neurons, had cell bodies with Dogiel type II morphology and their processes supplied networks of varicose processes around other nerve cells. It is concluded that each functionally defined type of submucosal neuron has a characteristic morphology and that intrinsic primary afferent neurons synapse with secretomotor neurons to form monosynaptic secretomotor reflex circuits. Anat Rec Part A 272A: 475-483, 2003.

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Localisation of NK1 receptor immunoreactivity to neurons and interstitial cells of the guinea-pig gastrointestinal tract

et al. 1996

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Tachykinins, including substance P, neurokinin A, and neuropeptides K and gama, are expressed widely in the peripheral nervous system where they affect smooth muscle contraction, exocrine gland secretion, vascular permeability, and neurotransmission. Substance P, the preferred ligand for the NK1 receptor, is found in high concentrations in the enteric nervous system. In the present study, the localisation and distribution of the NK1 receptor was studied throughout the gastrointestinal tract of the guinea-pig by using a polyclonal antiserum raised against the C-terminal 15 amino acids of the NK1 receptor. Co-localisation with other neuronal markers was examined in the ileum. Nerve cell bodies reactive for the NK1 receptor were found in the myenteric plexus of all regions and the submucous plexus of the small and large intestines. In the small intestine, the interstitial cells of Cajal were also immunoreactive. Immunoreactivity was largely confined to cell surfaces. Almost all immunoreactive myenteric nerve cells had Dogiel type I morphology, and most of these were immunoreactive for nitric oxide synthase, a transmitter of inhibitory neurons to the muscle and of descending interneurons. Neuropeptide Y-containing secretomotor neurons in the submucous and myenteric plexuses also exhibited NK1 receptor immunoreactivity. NK1 receptors were present on a minority of tachykinin immunoreactive neurons of submucous ganglia. The results suggest that receptors on the longitudinal muscle might not be conventional NK1 receptors, that excitation of the circular muscle of the ileum is indirect, perhaps via the interstitial cells of Cajal, and that enteric inhibitory neurons may be excited via NK1 receptors.

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Neurochemically similar myenteric and submucous neurons directly traced to the mucosa of the small intestine

Cited by 185 publications

References 19 publications

Choline Acetyltransferase (ChAT) Immunoreactivity in Paraffin Sections of Normal and Diseased Intestines

Choline Acetyltransferase (ChAT) Immunoreactivity in Paraffin Sections of Normal and Diseased Intestines

Morphologies and projections of defined classes of neurons in the submucosa of the guinea‐pig small intestine

Localisation of NK1 receptor immunoreactivity to neurons and interstitial cells of the guinea-pig gastrointestinal tract

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