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This study reports on changes in CART-like immunoreactive (CART-LI) nerve structures in the porcine descending colon during chemically driven inflammation and after axotomy. The distribution pattern of CART-LI nerve structures was studied using doublelabeling immunofluorescence technique in the circular muscle layer, myenteric (MP), outer submucous (OSP) and inner submucous plexuses (ISP) and also in the mucosal layer of the porcine descending colon in physiological conditions as well as under pathological factors. In the control animals, CART-LI perikarya have been shown to constitute 5.11% ± 0.64, 4.03% ± 1.17 and 0.05% ± 0.04 in MP, OSP and ISP, respectively. Changes in CART-immunoreactivity depended on the pathological factor and the part of the enteric nervous system (ENS) studied. Numbers of CART-LI perikarya amounted to 2.77% ± 0.64, 2.60% ± 0.36 and 0.26% ± 0.19 during chemically-induced colitis and 3.04% ± 0.88, 2.46% ± 0.8 and 0.43% ± 0.09 after axotomy in MP, OSP and ISP, respectively. Both studied pathological processes also caused an increase in the number of CART-LI nerve fibers in the circular muscle as well as in the mucosal layer.
Recent decades has brought significant advances in our knowledge of the chemical coding and function of enteric neurons. Calcium ions are important second messenger involved in many aspects of neuron physiology. In the present study, we analyzed immunohistochemically the presence of calcium binding proteins (calretinin and calbindin) in various subpopulations of enteric neurons from the ovine duodenum. Ten percent of submucous neurons were immunoreactive (IR) to calretinin. The presence of calretinin was not detected in myenteric neurons. Calretinin-expressing nerve fibres were found in both myenteric and submucous ganglia, between the circular and longitudinal smooth muscle layers and in the lamina muscularis mucosae. Calretinin-IR submucous neurons did not exhibit the presence of SP, NPY and VIP. Co-localization of calretinin and serotonin was found only in a small number of submucous neurons. Calbindin was expressed in 35% of myenteric neurons and in 60% of submucous neurons. Nerve fibres containing calbindin were localized in myenteric and submucous ganglia where they frequently formed basket-like formations. Calbindin-positive nerve fibres emerging from myenteric ganglia ran between the circular and longitudinal smooth muscle layers. Immunoreactivity to calbindin was also visualized in the lamina muscularis mucosae, around mucosal glands and blood vessels. None of calbindin-IR myenteric neurons revealed immunoreactivity to SP, NPY, VIP and serotonin. Virtually all calbindin-expressing submucous neurons were SP-positive. In moderate numbers of submucous perikarya, co-incidence of calbindin and NPY, calbindin and VIP or calbindin and serotonin was observed. We conclude that in the ovine duodenum, the expression of calretinin and calbindin is species specific. Co-localization studies and distribution patterns indicate that in the duodenum of the sheep, calretinin and calbindin may be present in several functional subclasses of enteric neurons.
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