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Retrograde neuronal tracing, using fast blue, in combination with a single-labelling immunofluorescence technique, was applied to determine whether somatostatin (SOM) participates in sensory innervating of the porcine adrenal glands in physiological conditions and after adrenalectomy. In control animals, SOM-like immunoreactive neurons comprised 7.0 ± 0.7% of adrenal gland-projecting cells in dorsal root ganglia (DRG) at neuromeres Th6-7 and 6.5 ± 1.2% at neuromeres Th 12-14. After adrenalectomy the percentage of SOM-positive DRG cells considerably increased and attained the level of 44.7 ± 2.5% at neuromeres Th6-7 and 36.6 ± 1.7% at neuromeres Th12-14. The obtained results demonstrate that SOM is not only a neuromediator within sensory neurones supplying the porcine adrenal glands, but also suggest the role of this substance during repairing processes within the nervous system after adrenalectomy.
This is the first study to examine zearalenone-(ZEN) induced changes in the immune system of the ileum and substance P-(SP-) and vasoactive intestinal peptide-(VIP-) immunoreactive nerve fibers in the mucosa, which participate in the regulation of intestinal functions under physiological conditions and during pathological processes. The aim of this study was also to identify potential relationships between selected immune and neural elements in ileal Peyer's patches in pigs that were and were not exposed to ZEN. The experiment was performed on 10 prepubertal gilts divided into two groups: the experimental group (n=5) where ZEN was administered at 0.1 mg kg-1 feed day-1 for 42 days, and the control group (n=5) which was administered a placebo. The tissue levels of cytokines were determined by enzyme-linked immunosorbent assay which revealed elevated concentrations of IL-12/23 40p and IL-1 ß in animals exposed to ZEN. Flow cytometry revealed a lower percentage of CD21+ lymphocytes in pigs exposed to ZEN in comparison with control animals. The tissue levels of neuropeptides were evaluated in the dot blot procedure which demonstrated higher concentrations of VIP and SP in experimental pigs. In experimental animals, numerous VIP-like immunoreactive processes were observed, and SP-immunoreactive nerve fibers formed a very dense network. Our results demonstrate for the first time that ZEN can modify the chemical coding of nerve structures in the gastrointestinal system. Those modifications can be attributed to ZEN's impact on estrogen receptors or its pro-inflammatory properties, and they reflect changes that take place in the nervous system at the transcriptional, translational and metabolic level.
The distribution pattern of CART- like immunoreactive (CART-LI) nerve structures was studied by a double immunofluorescence technique in the circular muscle layer, myenteric (MP), outer submucous (OSP), and inner submucous plexuses (ISP), as well as in the mucosal layer of porcine descending colon under physiological conditions and during proliferative enteropathy (PE). In control animals, CART-LI perikarya have been shown to constitute 3.18 ±0.51%, 3.44 ±0.6%, and 3.33 ±0.72% in MP, OSP, and ISP, respectively. PE caused a decrease in the number of CART - LI neurons in MP and ISP. In OSP, the observed changes were not statistically significant. During PE, the number of CART-LI perikarya amounted to 2.18 ±0.27%, 3.07 ±0.06%, and 0.07 ±0.02% within MP, OSP, and ISP, respectively. Moreover, PE caused an increase in the number of CART-LI nerve fibers in the colonic circular muscle and mucosal layers as well as in MP. This study describes for the first time changes in CART-LI nerve structures of the porcine descending colon during Lawsonia intracellularis infection, and suggests the participation of this neuropeptide in the regulation of gut functions, not only under physiological conditions, but also during pathological processes.
Changes in the density of mucosal pituitary adenylate cyclase activating peptide-27 -like immunoreactive (PACAP-27 - LI) nerve fibers within various parts of the canine gastrointestinal (GI) tract during inflammatory bowel disease (IBD) were investigated. The distribution of nerves were studied, using a single-labelling immunofluorescence technique, in the mucosal layer of canine stomach, duodenum, jejunum, and descending colon. Canine IBD caused an increase in the density of PACAP- 27-LI mucosal nerves in all studied parts of GI tract. The results suggest that PACAP in the nervous system may be involved in pathological processes during IBD.
The vasoactive intestinal peptide (VIP) and opioid family member Leu⁵-enkephalin (LENK) have already been established as playing independently significant roles in the functioning of the female genital tract. However, the mutual influence of both neuropeptides on female genital function has not been examined until now. Therefore, the aim of this study was to compare the distribution of VIP- and/or LENK-immunoreactive (IR) structures throughout the female genital tract of the pig. Immunohistochemical examination revealed that the great majority of the immunopositive structures co-expressed both peptides. Nevertheless, a small population of exclusively VIP- or LENK-IR processes and perikarya were also distinguished. The muscular layer of the organs examined revealed the greatest density of VIP- and/or LENK-IR nerve fibers. The mucosa of the ampulla, isthmus, cervix and vagina was supplied with a moderate number of single labeled LENK-IR processes, while exclusively VIP-IR fibers were found mainly in vaginal mucosa. The infundibulum was found to be poorly supplied with single labeled VIP- or LENK-IR fibers. The paracervical ganglion (PCG), the expected source of VIP- and/or LENK-IR nerve fibers innervating the organs under investigation, has been found to contain double labeled LENK-/VIP-IR as well as single labeled VIP-IR perikarya. The great number of specific co-localization between VIP and LENK in nerve processes of the porcine female genital organs may indicate a functional regulatory interaction between the neuropeptides studied, requiring further study.
The aim of the study was to determine the existence and co-existence patterns of VIP and NPY in neurons and nerve fibers of porcine lumbar-sacral sympathetic chain ganglia. The studied ganglia were fixed with 4% buffered paraformaldehyde (perfusion) and then labeled by means of double-immunofluorescence using a mixture of antibodies cultivated in different species. The highest number of NPY-positive cells was observed in the lumbar ganglia and diminished in the direction of the caudal, where only single neurons were observed. In contrast, a different pattern of distribution was observed for VIP-positive neurons, whose number was higher in the more caudally located ganglia. Two populations of VIP-positive neurons could be distinguished: single, showing strong immunofluorescence and often with visible processes, located in the central part of the ganglia and a second population, composed of clusters of 4-8 cells and often co-localizing NPY. VIP-positive nerve fibers surrounded both NPY+ neurons and neurons lacking either NPY and/or VIP. The presence of a small number of NPY-positive neurons exhibiting very weak immunofluorescence in more caudally located SChG could suggest a "switch" of neuromediators produced there. An increase in the percentage of non-noradrenergic sympathetic neurons in more caudally located SChG may thus be implicative for a specific innervation pattern of target tissues of these ganglia.
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