Search

In this study, we report the distribution of orexin A (OXA), orexin B (OXB), and orexin receptor (OX2R) immunoreactive (ir) cells in the hypothalamus and gastrointestinal tract of Oncorhynchus mykiss fed diets with different dietary fatty acid compositions. Trout were fed five iso-energetic experimental diets containing fish oil, or one of four different vegetable oils (olive, sunflower, linseed, and palm oils) as the added dietary lipid source for 12 weeks. OXA, OXB, and OX2R immunoreactive neurons and nervous fibers were identified in the lateral and ventro-medial hypothalamus. OXA, OXB, and OX2R ir cells were found in the mucosa and glands of the stomach and in the mucosa of both the pyloric cecae and intestine. OX2R ir cells were localized in the mucosa layer of both the pyloric cecae and intestine. These immunohistochemical (IHC) results were confirmed via Western blotting. Antibodies against preproorexin (PPO) crossreacted with a band of ∼16 kDa in the hypothalamus, stomach, pyloric cecae, and intestine. Antibodies against OX2R crossreacted with a band of ∼38 kDa in the hypothalamus, pyloric cecae, and intestine. The presence and distribution of OXA, OXB, and OX2R ir cells in the hypothalamus and gastrointestinal tract did not appear to be affected by dietary oils. The presence of orexin system immunoreactive cells in the stomach, pyloric cecae, and intestine of rainbow trout, but not in the enteric nervous system, could suggest a possible role of these peptides as signaling of gastric emptying or endocrine modulation, implying a main local action played by orexins.

In this study, we report nesfatin-1 immunoreactivity in the gastrointestinal tract of Casertana breed pig. The newly discovered anorexigenic peptide nesfatin-1 has been shown to possess physiological relevance in regulating food intake and energy homeostasis at a central level, although evidence has been accumulating that it may also play important functions at a more local gastroenteric level. Nesfatin-1 immunoreactive endocrine cells have been detected in the gastric fundus and ileocecal valve. Nesfatin-1 immunopositive neurons and nerve fibers have been observed mainly in the enteric plexuses. Western blot analysis confirmed the immunohistochemical observations, showing immunoreactive bands in all analyzed gastrointestinal tracts with the exception of the rectum. Nesfatin-1 immunodetection in the swine digestive system reinforces the importance of the role played by nesfatin-1 at the gastrointestinal level and sustains the necessity to study the role of this peptide in the regulation of food intake in farm species for which weight gain is essential for optimizing production.

Summary The presence of neuroendocrine (NE) cells producing biogenic amines and hormonal peptides has been investigated in the vestibular glands of the genital tracts of cows and pigs using immunohistochemistry. NE cells containing chromogranin A-, serotonin-, cholecystokinin- and somatostatin-immunoreactive material were found in both major and minor vestibular glands. Such cells were numerous, scattered in the acini and excretory duct epithelium, small in size and rounded, triangular or bipolar in shape. The function of the NE vestibular cells has been related to the secretory activity of the glands and to a sexual climax induction mechanism involving the stimulation of 5HT 3 receptors of vestibular nociceptor nerve fibers. The role of NE cells in small cell carcinomas of the vestibular glands is a topic for further investigation owing to possible parallelism between this type of tumor and the small cell carcinoma of the human prostate.

Orexin receptor 1 (OX₁R) and orexin receptor 2 (OX₂R) are two G-protein-coupled receptors that bind their ligands, orexin A (OXA) and B (OXB), with different affinities. The male genital system represents an important target for OXA, which appears to play a role in the control of steroidogenesis and germ cell development in the testis. It is known that among domestic breeding animals, in the boar the number of Leydig cells is very high and OXA appears to have stimulatory activity on testosterone production. In this study, we aimed to evaluate the presence of OX₁R in the boar testis in order to extend our knowledge concerning the distribution and a potential functional role of the orexinergic system in the male reproductive tract of farm animals. The presence of OX₁R immunopositive cells in seminiferous tubules of the boar testis enables us to hypothesize a possible role of OXA on male germ cells cycle in pig. Further investigations, involving functional and ultrastructural analysis, may contribute to our understanding of the role of orexins in the boar genital system.

This study provides an immunohistochemical approach to the expression of leptin in the gastrointestinal tract of the monogastric South American sea lion (Otaria flavescens), and the poligastric bottlenose dolphin (Tursiops truncatus). The specific organization of the gastrointestinal tract is examined in relation to the neuroendocrine regulation of the gut exerted by leptin. In the South American sea lion some leptin-like-immunoreactive (ir) cells, and endocrine type cells, were found in the pit of gastric mucosal folds and in the epithelium of duodenum as well as numerous neurons were detected in the submucosal and myenteric plexuses of the stomach. In the bottlenose dolphin, many leptin-like-ir cells, and exocrine type cells, were identified in the mucosal layer of the main stomach as well as several neurons and nervous fibers were detected in nervous plexuses of main stomach, pyloric stomach, proximal, and middle intestine. Our data suggest that the distribution of leptin-like peptides is similar in the two species, notwithstanding the different anatomical organization of the gastrointestinal apparatus of South American sea lion and bottlenose dolphin. These findings "suggest" the presence of a basal plan in the regulation of food intake, body weight, energy balance and of the gastrointestinal functions in general also in marine mammals with different and specific feeding habits.

KEY WORDS ghrelin; Carassius auratus; Dicentrarchus labrax; immunohistochemistry;gastrointestinal tractABSTRACT The occurrence and localization of ghrelin peptide in the gastrointestinal tract ofCarassius auratus and Dicentrarchus labrax, two fish species that exhibit different feeding behav-ior, different habitats, and different anatomical organizations of the gastroenteric tract, wereexamined by immunohistochemical methods and western blotting analysis. All of the gastrointesti-nal segments studied displayed immunohistochemical localizations of ghrelin peptide. Numeroussingle or clustered immunoreactive cells were found along the gastric folds, particularly in the py-loric region of Dicentrarcus labrax, whereas scattered ghrelin immunoreactive cells were observedin the intestinal epithelium of both fish species. Double immunolabeling PGP 9.5/ghrelin demon-strated the localization of ghrelin peptide also in nerve fibers and neuronal cells of the submucosaland myenteric plexuses, often in association with vascular structures. Western blotting analysisconfirmed the presence of ghrelin peptide in the gatrointestinal tract of both species studied, whosemolecular weight was similar to that of the corresponding mammalian prepro-ghrelin. The findingscould support the hypothesis that this peptide is an important appetite regulator in fish and couldconfirm the presence of the ghrelin peptide, starting from its precursor proteins, in the gastrointes-tinal tract of the goldfish and the sea bass. Microsc. Res. Tech. 00:000–000, 2009.

Orexin A (oxA) and orexin B are recently discovered peptides derived from the proteolytic cleavage of the common precursor prepro-orexin. They bind two G protein-coupled receptors, defined orexin 1 (ox1R) and orexin 2 receptor. Both peptides are highly expressed in the lateral hypothalamic area of the brain and are involved in the regulation of many functions of the body, the best investigated of which is food intake. Recent data described the presence of orexins in peripheral organs such as the adrenal glands, stomach, bowel, pancreas, and testis. Here, we report the detection of oxA and ox1R in the exocrine and endocrine cytotypes of the cattle urethroprostatic complex by using immunohistochemistry. The expression of prepro-orexin and ox1R mRNA transcripts in the prostatic tissue was assessed by reverse-transcriptase polymerase chain reaction, while the presence of both the proteins in the tissue was confirmed by Western blotting analysis. Our findings provide the first evidence for the presence of oxA and ox1R in the urethroprostatic complex of the cattle and demonstrate that both proteins are locally synthesized, thus suggesting a role for oxA on both physiological and pathological functioning of the complex.