Orexin and orexin receptor like peptides in the gastroenteric tract of Gallus domesticus: An immunohistochemical survey on presence and distribution.

This study reports the immunohistochemical localization and distribution of orexin A and B-like and their receptors-like peptides in the gastroenteric tract of chicken. The immunoreactivity is distributed in endocrine cells, nerve fibers and neurons, both in the stomach and intestine, and shows a discrete conformity with the data till now reported for Mammals. Our study suggests a possible participation of orexin-like peptides in the modulation of chicken gastroenteric activities and the preservation of their main distribution compared to Mammals. Western blot analysis has confirmed the presence of prepro-orexin and both receptors in the examined tissues. This survey represents the first evidence of the presence of orexin-like peptides in the gastroenteric tract of non mammalian species, and the results could help to better understand the alimentary control and body weight in domestic birds, which are of relevance to determine the productive factors in breeding animals. This study might also serve as a baseline for future experimental studies on the regulation of the gastroenteric functions in non mammalian Vertebrates.

TRK neurotrophin receptor-like proteins in the kidney of frog (Rana esculenta) and lizard (Podarcis sicula): an immunohistochemical study.

The present study investigates the occurrence of Trk-like neurotrophin receptor proteins in the lizard and frog kidney. In lizard rare TrkB-like immunoreactive cells in intermediate and distal tubules were found. TrkC-like immunoreactive cells were numerous in collecting tubules and became less numerous in collecting ducts. No TrkC-like immunoreactivity was detected in the ureteric duct. In the frog, we observed numerous TrkC-like immunoreactive cells in collecting tubules and ducts while they were scattered among negative epithelial cells in the wolffian duct. TrkB- and TrkA-like immunoreactivity was never found. Western blot analysis demonstrated that the frog and lizard kidney contains TrkC-like protein; TrkB-like protein was present only in the lizard kidney. These results demonstrate for the first time the occurrence of Trk-like proteins in the kidney of amphibians and reptiles, and aid in the assessment of the role of Trk receptor-like proteins in the kidney physiology of vertebrates.

Neurotrophin-like immunoreactivity in the gut of teleost species.

By means of immunochemistry and immunohistochemistry, we investigated in the gut of teleostean species the presence and localization of three neurotrophins: nerve growth factor (NGF), brain derived neurotrophic factor (BDNF) and neurotrophin (NT)-3. In all studied species both NGF- and NT-3-like immunoreactivity (IR) were present in the enteric nervous system, while BDNF-like IR was never detected. More in particular, both NGF and NT-3-like IR were detected in neurons of small and large intestine, while only NT3-like IR was also observed in stomach plexuses. Furthermore, Western blot analysis revealed the presence of molecules immunoreactive to NGF and NT-3, which weight were very similar to those of mammalian corresponding neurotrophins. These results extend to teleost species the presence and distribution of NGF- and NT-3-like IR in the enteric nervous system, suggesting a well-preserved presence of these substances in the gut during vertebrate phylogenesis.

S100-like protein in the goldfish (Carassius auratus) kidney. An immunohistochemical study.

The presence and distribution of S100-like protein in the goldfish (Carassius auratus L.) kidney has been studied by the use of immunohistochemical and histochemical methods. Simple immunohistochemistry (peroxidase anti-peroxidase method) was carried out with a polyclonal antibody against a mixture of both S100alpha and S100beta proteins. In order to confirm the cell-type containing S-100-like immunoreactivity, the colocalization of S-100-like protein immunoreactivity with periodic acid-Schiff (PAS) reaction was investigated by using double staining with indirect immunofluorescence and PAS histochemistry. S100-like immunoreactivity was detected only in juxtaglomerular cells located in the renal arterial branch and never on afferent arterioles. No immunoreactivity was observed in other tracts of the nephron or in the interstitial cells. Double staining confirmed that S-100-like immunoreactivity and PAS reactivity were colocalized in juxtaglomerular cells. These findings are the first regarding the presence and distribution of S100-like protein in the teleost kidney; they add a new member to the list of extra-neural S100-like-containing cell types and confirm that the antigen cannot be regarded as nervous-system-specific. In addition, a concentration of S100-like immunoreactivity in juxtaglomerular cells suggests the presence of S100-like calcium-binding protein-mediated activities in these cell types.

The teleost kidney expresses Trk neurotrophin receptor-like proteins.

Neurotrophins are involved in the development and maintenance of some neuronal populations, acting through tyrosine protein kinase (Trk) receptors, TrkA, TrkB and TrkC. In addition, recent studies have demonstrated the occurrence of Trk receptors in a wide variety of adult and embryonic non-neuronal tissues in vertebrates, including kidney. Nevertheless no data are available on Trk proteins distribution in teleost kidney. The present study, by using an immunoperoxidase technique, analyses Trk receptor protein distribution in the kidney of three species of freshwater and saltwater teleost. TrkA-like immunoreactivity was the only one detected in all examined species, while TrkB-like immunoreactivity was completely absent and TrkC-like immunoreactivity was detected only in goldfish. The TrkA immunoreactive cells were mainly localised in the collecting duct system, as this system was the only one to also display TrkC. Such data could provide new clues to Trk in fish and aid assessment of the role of Trk protein receptors during vertebrate evolution.

PACAP (pituitary adenylate cyclase-activating peptide)-like immunoreactivity in the gill arch of the goldfish, Carassius auratus: distribution and comparison with VIP.

Pituitary adenylate cyclase-activating peptide (PACAP) is a novel vasoactive intestinal peptide (VIP)-like peptide isolated from ovine hypothalamus. It is present in neuronal elements of a number of peripheral organs. We have examined whether PACAP occurs in the gill arch of Carassius auratus L. in which our recent studies have shown the presence of VIP-like peptide. Immunohistochemistry has revealed PACAP-like immunoreactivity in the anterior branches of the post-trematic glossopharyngeal and vagus nerves. PACAP-immunoreactive nerve cell bodies and fibers are present in connective tissue on the oral side of the gill arch. Colocalization studies carried out by the application of double immunofluorescence show that a PACAP-like peptide coexists with VIP in the same nerve cell bodies and fibers. The localization pattern of PACAP in the gill arch of goldfish suggests its possible involvement in the regulation of secretory activities.

Vasoactive intestinal polypeptide immunoreactive nerves in the gill arch of teleost fish, Carassius auratus L.

The localization of vasoactive intestinal polypeptide (VIP)-immunoreactive (ir) nerve cell bodies and fibers has been studied in the gill arches of goldfish (Carassius auratus, L.) using the peroxidase-antiperoxidase (PAP) immunohistochemical method. It was found that VIP-ir nerve cell bodies are localized in connective tissue on the oral side of the gill arch; these cells were present as single cells, in couples or as small clusters. Moreover, a dense network of VIP-ir fibers was observed beneath the lining epithelium of the raker cushion. The possible involvement of this peptide in mucus secretion in the gill arches of teleost is discussed.

VIP-like immunoreactive cells in the kidney of goldfish (Carassius auratus).

By immunohistochemistry VIP-like immunoreactive cells were observed in the second segment of some renal proximal tubules of Carassius auratus. Immunoreactivity displayed a granular appearance all over the cells. VIP-like immunoreactive cells showed a shape of an over-turned flask whose globe-like part looks like the tubular lumen. No immunoreactive material was observed in other tracts of the nephron and in the interstitial cells. The presence of VIP-like immunoreactive cells is discussed in relation to their possible function.