Neuropeptide tyrosine-like immunoreactivity (NPY-LI) in ganglion neurons in the adrenal gland of the flat snake (Waglerophis merremii)

In previous morphological and histochemical studies on the adrenal gland of the flat snake, no data demonstrating the existence of ganglion neurons has been reported. The aim of this paper was therefore to establish the presence of ganglion neurons in the adrenal gland of the flat snake Waglerophis merremii and, further to study their chemical phenotype using immunohistochemistry. Our results showed the presence of cells which were immunolabelled with the neuronal marker neurofilament 10 and were thus identified as large ganglion neurons. These cells were localized in the dorsal ribbon of the gland, suggesting a noradrenergic phenotype, exhibited long processes with a longitudinal direction and co-expressed neuropeptide tyrosine- (NPY) and tyrosine hydroxylase-like immunoreactivities (-LI).In addition, NPY-immunoreactive (-IR) fibers were recognized with a wide distribution throughout the gland whereas vasoactive intestinal polypeptide (VIP)-IR fibers were only observed between clusters of cortical and adrenergic chromaffin cells. No cells containing VIP-LI were detected within the gland. Based on their histochemical phenotype, ganglion cells containing NPY and TH could correspond to ganglion neurons type I of the rat. The possible absence of type II ganglion neurons in the adrenal gland of the snake is discussed

An immunohistochemical study of temperature-related changes in galanin and nitric oxide synthase immunoreactivity in the hypothalamus of the toad.

Galanin (GAL) and nitric oxide synthase (NOS) have been implicated in the control of thermogenesis in mammals. An experimental protocol was designed to determine whether or not the expression of these molecules in the hypothalamus of the toad could be related to environmental temperature changes. Exposure of the animals to low temperature increased the number and intensity of NOS-positive neurons in the magnocellular hypothalamic region, in contrast to a weak immunoreactivity observed in control animals kept in a natural environment at a spring-summer temperature (23-27 degrees C). Also a significantly higher number of GAL-immunoreactive (-IR) cells was observed in the preoptic area as compared to that observed in controls, while no difference in the intensity of GAL immunostaining intensity was detected. These results show a temperature-related expression of GAL and NOS in the hypothalamus and preoptic area of the toad. The results suggest a possible role of GAL and NOS in the regulation of hibernation in these animals.

Neuropeptide tyrosine-like immunoreactivity (NPY-LI) in ganglion neurons in the adrenal gland of the flat snake (Waglerophis merremii).

In previous morphological and histochemical studies on the adrenal gland of the flat snake, no data demonstrating the existence of ganglion neurons has been reported. The aim of this paper was therefore to establish the presence of ganglion neurons in the adrenal gland of the flat snake Waglerophis merremii and, further to study their chemical phenotype using immunohistochemistry. Our results showed the presence of cells which were immunolabelled with the neuronal marker neurofilament 10 and were thus identified as large ganglion neurons. These cells were localized in the dorsal ribbon of the gland, suggesting a noradrenergic phenotype, exhibited long processes with a longitudinal direction and co-expressed neuropeptide tyrosine- (NPY) and tyrosine hydroxylase-like immunoreactivities (-LI). In addition, NPY-immunoreactive (-IR) fibers were recognized with a wide distribution throughout the gland whereas vasoactive intestinal polypeptide (VIP)-IR fibers were only observed between clusters of cortical and adrenergic chromaffin cells. No cells containing VIP-LI were detected within the gland. Based on their histochemical phenotype, ganglion cells containing NPY and TH could correspond to ganglion neurons type I of the rat. The possible absence of type II ganglion neurons in the adrenal gland of the snake is discussed.

Presence of DOPA decarboxylase and its localisation in adult rat pancreatic islet cells.

The aim of this work was to investigate the possible presence of DOPA decarboxylase (DDC) in endocrine cells of adult rat pancreas. Islet peptide hormones (insulin, glucagon, and somatostatin), as well as DDC, were detected immunohistochemically using the double-immunofluorescence technique and specific antibodies. DDC-like immunoreactivity was present in cytoplasmic granules within endocrine cells located at islet peripheries in a distribution consistent with islet localisation of A cells. Moreover, these same cells stained positively with glucagon antibody. As DDC is an enzyme specifically involved in catecholamine synthesis, insular cells must possess the capacity to elaborate this class of hormone at least up to the dopamine-decarboxylation step. Thus, after further metabolic processing either in A cells or elsewhere, endogenously-synthesised islet catecholamines may be released and participate in paracrine regulation of insulin secretion.

Growth hormone inhibits the hypophysectomy-induced expression of galanin in hypothalamic neurons of the toad (Bufo arenarum hensel).

The expression of the neuropeptide galanin was analyzed by immunohistochemistry in magnocellular and preoptic hypothalamic neurons of toads following hypophysectomy (HPX) and pars distalectomy (PDX). There was a marked increase in the galanin-like immunoreactive expression in magnocellular hypothalamic cells 3 days after HPX, followed by a decrease to normal levels after 7 days. No changes in the expression of galanin were detected after PDX in these neurons when compared to controls. Moreover, 7 days after HPX or PDX the number of cells expressing galanin was significantly increased in the preoptic area, where numerous intraependymal cells were intensely immunoreactive. The hypophysis grafts into the hind limb in HPX or PDX animals prevented increased galanin-like immunoreactivity in preoptic cells but not in magnocellular neurons. Similarly, PDX toads given growth hormone showed no GAL-LI in the intraependymal preoptic cells. These results suggest the presence of a region regulation of galanin expression in the preoptic area by hypophyseal hormones, in particular growth hormone.

Galanin-like immunoreactive expression in the central nervous system of the toad.

The distribution of galanin (GAL)-like immunoreactivity (-LI) was studied in the CNS of the toad (Bufo arenarum Hensel). Tissue sections were incubated with antibodies directed toward rat or porcine GAL and processed either for the avidin-biotin complex, or for the indirect immunofluorescence techniques. In the telencephalon GAL-immunoreactive (-IR) perikarya were observed in the ventral part of the striatum and in the septal accumbens nuclei. Immunopositive neurons were also observed in the medial amigdala with some intermingled cells between the fibers of the anterior commissure. Numerous GAL-IR perikarya were present along the rostrocaudal medial preoptic nucleus. Occasionally lightly immunoreactive cells were detected in the magnocellular region. The most numerous accumulation of GAL-IR cells was present in the ventral hypothalamus around the infundibular region, in the posterior tubercle and in the nucleus of the paraventricular organ. Immunostained cells were also present in the pretectal gray, solitary nucleus, gracil nucleus and in the spinal cord in the intermediate gray and in large motoneurons of the ventral horn. The widespread distribution found of GAL-LI suggests that GAL in the toad, as well as in mammalian species, may serve a variety of functions with a preponderant role in neuroendocrine processes. A role for GAL as a trophic factor in the brain of the toad is also suggested.

Galanin-like immunoreactive expression in the central nervous system of the toad

The distribution of galanin (GAL)-like immunoreactivity (-LI) was studied in the CNS of the toad (Bufo arenarum Hensel). Tissue sections were incubated with antibodies directed toward rat or porcine GAL and processed either for the avidin-biotin complex, or for the indirect immunofluorescence techniques. In the telencephalon GAL-immunoreactive (-IR) perikarya were observed in the ventral part of the striatum and in the septal accumbens nuclei. Immunopositive neurons were also observed in the medial amigdala with some intermingled cells between the fibers of the anterior commissure. Numerous GAL-IR perikarya were present along the rostrocaudal medial preoptic nucleus. Occasionally lightly immunoreactive cells were detected in the magnocellular region. The most numerous accumulation of GAL-IR cells was present in the ventral hypothalamus around the infundibular region, in the posterior tubercle and in the nucleus of the paraventricular organ. Immunostained cells were also present in the pretectal gray, solitary nucleus, gracil nucleus and in the spinal cord in the intermediate gray and in large motoneurons of the ventral horn. The widespread distribution found of GAL-LI suggests that GAL in the toad, as well as in mammalian species, may serve a variety of functions with a preponderant role in neuroendocrine processes. A role for GAL as a trophic factor in the brain of the toad is also suggested.

Immunohistochemical localization of neurotensin in a subpopulation of noradrenergic chromaffin cells of the adrenal gland of the flat snake (Waglerophis merremii).

The distribution of neurotensin (NT)-like immunoreactivity (LI) in the adrenal gland of the snake Waglerophis merremii has been examined immunohistochemically. Double staining, combining NT with tyrosine hydroxylase (TH) or calcitonin gene-related peptide (CGRP) antibodies and TH with CGRP antibodies, was also carried out. Results were analyzed by conventional and by confocal fluorescence microscopy. Immunostaining revealed a subpopulation of chromaffin cells containing NT-LI within the dorsal noradrenergic ribbon. In addition, there were some NT-immunoreactive (IR) fibers in this region. NT immunoreactivity was not present within adrenergic chromaffin cells or in cortical tissue. Double staining revealed CGRP-IR fibers innervating most of the chromaffin adrenergic cells. Within the dorsal noradrenergic ribbon, two groups of chromaffin TH-IR cells were present, one receiving a dense CGRP-IR innervation and another without contact with CGRP-IR terminals. The latter chromaffin cells displayed NT-LI. These results show, for the first time, the presence of a neuropeptide in chromaffin noradrenergic cells of a reptilian adrenal gland and open up the possibility that other peptides may also be present in these cells.

Histochemical study of chromaffin cells and nerve fibers in the adrenal gland of the flat snake (Waglerophis merremii).

The distributions of tyrosine hydroxylase (TH), protein gene product (PGP) 9.5, calcitonin gene-related peptide (CGRP), and peptide histidine isoleucine (PHI) have been examined immunohistochemically in the adrenal gland of the snake Waglerophis merremii. The morphology of chromaffin cells and the presence of ganglionic neurons in the gland revealed by means of the glutaraldehyde-silver technique and electron microscopy are also described. Two distinct types of TH-immunoreactive (-IR) cells are present in the dorsal noradrenergic ribbon: small chromaffin cells and a larger type identified as ganglionic neurons. Small, mostly round or fusiform cells often displayed long processes. Ganglionic cells, arranged in patches, had long processes entering the cortex of the gland. Chromaffin adrenergic cells, forming small groups of 4-7 cells, were scattered within the interrenal tissue and had a wide variety of shapes with processes that appeared to contact other chromaffin cells. Bundles of PGP 9.5-IR fibers occurred in the subcapsular zone of the adrenal gland with fibers entering the cortex and dorsal noradrenergic ribbon of the gland. Thick and thin TH-IR fibers were seen. Thick TH-IR fibers were nonvaricose and appeared to originate mainly in ganglionic neurons. Thin TH-IR fibers with small varicosities were numerous in the interrenal tissue and were frequently seen between clusters of adrenergic cells in close apposition to cortical cells and vessels. CGRP-IR fibers were present throughout the entire adrenal gland, whereas PHI-IR fibers had a preferential distribution in the interrenal tissue. Both CGRP- and PHI-IR fibers were closely associated with vessels and cortical cells.