Immunohistochemical analysis of neuron types in the mouse small intestine |
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Authors: | Zheng-Dong Qu Michelle Thacker Patricia Castelucci Mária Bagyánszki Miles L Epstein John B Furness |
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Institution: | Department of Anatomy & Cell Biology and Centre for Neuroscience, University of Melbourne, Parkville, Victoria, 3010, Australia. |
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Abstract: | The definition of the nerve cell types of the myenteric plexus of the mouse small intestine has become important, as more
researchers turn to the use of mice with genetic mutations to analyze roles of specific genes and their products in enteric
nervous system function and to investigate animal models of disease. We have used a suite of antibodies to define neurons
by their shapes, sizes, and neurochemistry in the myenteric plexus. Anti-Hu antibodies were used to reveal all nerve cells,
and the major subpopulations were defined in relation to the Hu-positive neurons. Morphological Type II neurons, revealed
by anti-neurofilament and anti-calcitonin gene-related peptide antibodies, represented 26% of neurons. The axons of the Type
II neurons projected through the circular muscle and submucosa to the mucosa. The cell bodies were immunoreactive for choline
acetyltransferase (ChAT), and their terminals were immunoreactive for vesicular acetylcholine transporter (VAChT). Nitric
oxide synthase (NOS) occurred in 29% of nerve cells. Most were also immunoreactive for vasoactive intestinal peptide, but
they were not tachykinin (TK)-immunoreactive, and only 10% were ChAT-immunoreactive. Numerous NOS terminals occurred in the
circular muscle. We deduced that 90% of NOS neurons were inhibitory motor neurons to the muscle (26% of all neurons) and 10%
(3% of all neurons) were interneurons. Calretinin immunoreactivity was found in a high proportion of neurons (52%). Many of
these had TK immunoreactivity. Small calretinin neurons were identified as excitatory neurons to the longitudinal muscle (about
20% of neurons, with ChAT/calretinin/± TK chemical coding). Excitatory neurons to the circular muscle (about 10% of neurons)
had the same coding. Calretinin immunoreactivity also occurred in a proportion of Type II neurons. Thus, over 90% of neurons
in the myenteric plexus of the mouse small intestine can be currently identified by their neurochemistry and shape. |
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Keywords: | Enteric nervous system Chemical coding Small intestine Neurofilaments Nitric oxide synthase Mouse (BalbC) |
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