Immunohistochemical characterization and quantitative analysis of neurons in the myenteric plexus of the equine intestine.
Abstract: The present study was performed on whole-mount preparations to investigate the chemical neuroanatomy of the equine myenteric plexus throughout its distribution in the intestinal wall. The objective was to quantify neurons of the myenteric plexus, especially the predominant cholinergic and nitrergic subpopulations. Furthermore, we investigated the distribution of vasoactive intestinal polypeptide and the calcium-binding protein calretinin. Samples from different defined areas of the small intestine and the flexura pelvina were taken from 15 adult horses. After fixation and preparation of the tissue, immunofluorescence labeling was performed on free floating whole-mounts. Additionally, samples used for neuropeptide staining were incubated with colchicine to reveal the neuropeptide distribution within the neuronal soma. The evaluation was routinely accomplished using confocal laser-scanning microscopy. For quantitative and qualitative analysis, the pan-neuronal marker anti-HuC/D was applied in combination with the detection of the marker enzymes for cholinergic neurons and nitrergic nerve cells. Quantitative data revealed that the cholinergic subpopulation is larger than the nitrergic one in several different locations of the small intestine. On the contrary, the nitrergic neurons outnumber the cholinergic neurons in the flexura pelvina of the large colon. Furthermore, ganglia are more numerous in the small intestine compared with the large colon, but ganglion sizes are bigger in the large colon. However, comparison of the entire population of neurons in the different locations of the gut showed no difference. The present study adds further data on the chemoarchitecture of the myenteric plexus which might facilitate the understanding of several gastrointestinal disorders in the horse.
Publication Date: 2008-10-07 PubMed ID: 18930715DOI: 10.1016/j.brainres.2008.09.070Google Scholar: Lookup
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- Journal Article
Summary
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The research paper investigates the chemical neuroanatomy of the equine myenteric plexus in the intestinal wall of horses. It aims to quantify neurons, specifically, the cholinergic and nitrergic neuronal subpopulations, and the distribution of certain proteins. The study results provide more insight into the functioning of the equine gastrointestinal tract, potentially aiding in understanding related disorders.
Research Method
- The researchers studied the myenteric plexus in the intestines of 15 adult horses, with samples taken from various defined areas of the small intestine and the flexura pelvina (a curve in the large intestine).
- After fixation and tissue preparation, immunofluorescence labeling was performed on the samples. This labels specific proteins in the tissue and allows them to be visualized under a microscope.
- For neuropeptide staining, to reveal neuropeptide distribution in the neuronal cells, samples were subjected to colchicine treatment.
- The evaluation was performed using confocal laser-scanning microscopy, a sophisticated imaging technique which allows high-resolution imaging of biological s
Quantitative and Qualitative Analysis
- The researchers used the pan-neuronal marker anti-HuC/D with marker enzymes for cholinergic and nitrergic neurons. This allowed them to differentiate and count these two types of nerve cells in the samples.
- The findings revealed that the cholinergic neuronal subpopulation is larger than the nitrergic one in several locations in the small intestine. However, in the flexura pelvina of the large colon, nitrergic neurons predominated.
- In comparison, there were more ganglia, groups of nerve cells, in the small intestine than in the large colon. However, the size of these ganglia was larger in the large colon.
Conclusions and Implications
- No specific difference was found in the total population of neurons in different locations of the gut. This suggests a certain degree of uniformity in the equine intestinal nervous system.
- The study added more data about the chemical makeup or chemoarchitecture of the equine myenteric plexus.
- This research might facilitate a greater understanding of various gastrointestinal disorders in horses, as the intestinal nervous system has a major role in controlling gut functions.
Cite This Article
APA
Freytag C, Seeger J, Siegemund T, Grosche J, Grosche A, Freeman DE, Schusser GF, Härtig W.
(2008).
Immunohistochemical characterization and quantitative analysis of neurons in the myenteric plexus of the equine intestine.
Brain Res, 1244, 53-64.
https://doi.org/10.1016/j.brainres.2008.09.070 Publication
Researcher Affiliations
- Paul Flechsig Institute for Brain Research, University of Leipzig, Jahnallee 59, D-04109 Leipzig, Germany.
MeSH Terms
- Animals
- Choline O-Acetyltransferase / metabolism
- Colon / anatomy & histology
- Colon / innervation
- Colon / metabolism
- Fluorescent Antibody Technique
- Ganglia, Autonomic / cytology
- Ganglia, Autonomic / metabolism
- Horses
- Immunohistochemistry
- Intestinal Mucosa / metabolism
- Intestine, Small / anatomy & histology
- Intestine, Small / innervation
- Intestine, Small / metabolism
- Intestines / anatomy & histology
- Intestines / innervation
- Microscopy, Confocal
- Myenteric Plexus / anatomy & histology
- Myenteric Plexus / cytology
- Myenteric Plexus / metabolism
- Neurons / cytology
- Neurons / metabolism
- Nitrergic Neurons / cytology
- Nitrergic Neurons / metabolism
- Nitric Oxide Synthase / metabolism
- Vasoactive Intestinal Peptide / metabolism
- Vesicular Acetylcholine Transport Proteins / metabolism
Citations
This article has been cited 8 times.- Yamada NO, Wenduerma, Senda T. Altered microbiota caused by disordered gut motility leads to an overactivation of intestinal immune system in APC1638T mice. Med Mol Morphol 2023 Sep;56(3):177-186.
- Brehmer A. Classification of human enteric neurons. Histochem Cell Biol 2021 Aug;156(2):95-108.
- Bombardi C, Rambaldi AM, Galiazzo G, Giancola F, Graïc JM, Salamanca G, Cozzi B, Chiocchetti R. Nitrergic and Substance P Immunoreactive Neurons in the Enteric Nervous System of the Bottlenose Dolphin (Tursiops truncatus) Intestine. Animals (Basel) 2021 Apr 8;11(4).
- Wilhelm M, Lawrence JJ, Gábriel R. Enteric plexuses of two choline-acetyltransferase transgenic mouse lines: chemical neuroanatomy of the fluorescent protein-expressing nerve cells. Brain Res Bull 2015 Feb;111:76-83.
- Chiocchetti R, Giancola F, Mazzoni M, Sorteni C, Romagnoli N, Pietra M. Excitatory and inhibitory enteric innervation of horse lower esophageal sphincter. Histochem Cell Biol 2015 Jun;143(6):625-35.
- da Silva MV, Marosti AR, Mendes CE, Palombit K, Castelucci P. Differential effects of experimental ulcerative colitis on P2X7 receptor expression in enteric neurons. Histochem Cell Biol 2015 Feb;143(2):171-84.
- Lu HL, Huang X, Wu YS, Zhang CM, Meng XM, Liu DH, Kim YC, Xu WX. Gastric nNOS reduction accompanied by natriuretic peptides signaling pathway upregulation in diabetic mice. World J Gastroenterol 2014 Apr 28;20(16):4626-35.
- Nagy JI, Urena-Ramirez V, Ghia JE. Functional alterations in gut contractility after connexin36 ablation and evidence for gap junctions forming electrical synapses between nitrergic enteric neurons. FEBS Lett 2014 Apr 17;588(8):1480-90.
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