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A Fasolo

Publications and source records attributed to A Fasolo.

At least 55 records · Page 3Linked to original sources
PubMed1996

Immunohistochemical investigation of gamma-aminobutyric acid ontogeny and transient expression in the central nervous system of Xenopus laevis tadpoles.

The ontogeny of the gamma-aminobutyric acid (GABA)-positive neurons in the brain of Xenopus laevis tadpoles was investigated by means of immunohistochemistry, using specific antibodies both against GABA and its biosynthetic enzyme, glutamate decarboxylase (GAD). The results obtained with the two antisera were comparable. The GABA system differentiates very early during development. At stages 35/36, numerous GABA-positive neurons were seen throughout the prosencephalon and formed two main bilateral clusters within the lateral walls of the forebrain that ran caudally toward the hindbrain. Other GABA-immunolabeled cell bodies, together with a conspicuous network of GABAergic fibers, were seen in the posterior hypothalamus. In the spinal cord, the lateral marginal zone was GABA-positive, as were Rohon-Beard neurons, interneurons, and Kolmer-Agdhur cells. A very rich GABA innervation was observed in the pars intermedia of the pituitary. At stage 50, plentiful immunopositive neurons and fibers were found in the telencephalic hemispheres, the diencephalon, and the mesencephalon (optic tectum and tegmentum). By stage 54, the number of GABA-immunoreactive neurons in the posterior hypothalamus had decreased, so that, at stage 58, there were very few GABA-labeled cell bodies in the dorsolateral walls of the infundibulum, despite a strong GABAergic innervation within the median eminence and the pars intermedia. From stage 58 to stage 66, the distribution pattern was very similar to that described in the adult X. laevis and in other amphibian species. These results point to transient GABA expression within the hypothalamus, possibly related to either 1) a naturally occurring cell death or 2) a phenotypic switch.

Animals↗
PubMed1996

Localization of diazepam-binding inhibitor-related peptides and peripheral type benzodiazepine receptors in the frog adrenal gland.

The adrenal gland of mammals contains high concentrations of peripheral-type benzodiazepine receptors (PBR) and diazepam-binding inhibitor (DBI), a polypeptide which acts as an endogenous ligand for PBR. The aim of the present study was to investigate the localization of DBI and PBR in the adrenal gland of the frog Rana ridibunda. Reverse transcription followed by polymerase chain reaction with specific primers for the frog DBI cDNA showed the presence of DBI mRNA in frog adrenal gland extracts. The cellular distribution of DBI and PBR was investigated using an antiserum against the octadecaneuropeptide DBI [33-50] (ODN) and antibodies against the 18-kDa isoquinoline binding protein subunit of PBR (IBP), respectively. ODN-like immunoreactivity was found in chromaffin cells and in Stilling s cells, but not in adrenocortical cells. IBP-like immunoreactivity was observed in chromaffin cells, in Stilling s cells and in a small proportion (11%) of steroid-secreting cells. The ODN- and IBP-immunoreactive materials were homogeneously distributed in the cytoplasm of chromaffin cells and concentrated at the periphery of large cytoplasmic vesicles in Stilling s cells. The proportion of ODN-positive Stilling s cells showed marked circannual variations with a maximum in July. Similarly, the proportion of IBP-positive Stilling s cells was 17 times higher in July than in December. These results indicate that, in the frog adrenal gland, DBI-related peptides and PBR are simultaneously expressed in chromaffin cells and Stilling s cells, suggesting that endogenous ligands for PBR may play a physiological role in the control of adrenal cell activity.

Adrenal Glands↗
PubMed1996

Ciliary neurotrophic factor constitutively expressed in the nervous system of transgenic mice protects embryonic dorsal root ganglion neurons from apoptosis.

Ciliary neurotrophic factor (CNTF) is a potent survival factor for several neuronal populations. It is expressed postnatally by Schwann cells in the peripheral nervous system and by some glial and neuronal cells in the central nervous system. We used the promoter of the neurofilament light chain gene to produce transgenic mice that express CNTF in neurons from the beginning of neuronal differentiation. These transgenic animals may represent a suitable model to identify neuronal cell types responsive to CNTF in vivo and to study the mechanism of action of this neurotrophic factor. We show that dorsal root ganglion neurons of transgenic mice expressing CNTF in neurons are protected from apoptosis during embryonic development: 40% of these cells undergo apoptosis between embryonic day 12.5 and postnatal day 5 in transgenic mice whereas 60% do so in control animals. However, protection from apoptosis does not result in an increase in the total number of neurons at the end of development. We discuss our results with regard to CNTF potentialities in vivo and the significance of programmed cell death during development.

Animals↗
PubMed1995

Distribution of two molecular forms of gonadotropin-releasing hormone (GnRH) in the central nervous system of the frog Rana ridibunda.

Two molecular forms of gonadotropin-releasing hormone (GnRH) have been recently characterized in the brain of the frog Rana ridibunda i.e. mammalian GnRH (mGnRH) and chicken GnRH-II (cGnRH-II). Using highly specific antisera against each form of GnRH, we have investigated the distribution of these two neuropeptides in the frog brain by the indirect immunofluorescence and the peroxidase-antiperoxidase techniques. mGnRH-immunoreactive cell bodies were restricted to a well defined region corresponding to the septal-anterior preoptic area. mGnRH-containing fibers projected through the ventral diencephalon and ended in the median eminence. In contrast, cGnRH-II-immunoreactive structures were widely distributed in the frog brain. In the telencephalon cGnRH-II-positive elements formed a ventromedial column extending from the olfactory bulb to the septal area, a pathway which corresponds to the terminal nerve. A dense accumulation of cGnRH-II-immunoreactive cell bodies was also found in the septal-anterior preoptic area; these neurons sent processes towards the median eminence via the hypothalamus. Double immunostaining revealed that, in this area, mGnRH- and cGnRH-II-like immunoreactivity co-existed in the same neurons. In the mid-diencephalon, numerous cGnRH-II-immunoreactive perikarya were found, surrounding the third ventricle, in the posterior preoptic and infundibular areas. Many of these neurons sent processes towards the ventricular cavity. More caudally, a dense population of cGnRH-II-immunoreactive perikarya was also observed in the nucleus of the paraventricular organ and the posterior tubercle. Dorsally, the thalamus, the tegmentum, the tectum and the granular layer of the cerebellum were richly innervated by cGnRH-II-positive fibers. In the medulla oblongata, numerous cGnRH-II-immunoreactive perikarya were seen in several cranial nerve nuclei. Ventrally, a dense plexus of immunoreactive fibers projected rostrocaudally into the spinal cord. The occurrence of mGnRH- and cGnRH-II-like immunoreactivity in the septal-anterior preoptic area and the hypothalamo-pituitary pathway supports the view that both peptides act as hypophysiotropic neurohormones. The widespread distribution of cGnRH-II-immunoreactive elements in the central nervous system of the frog strongly suggests that this peptide may also exert neuromodulator and/or neurotransmitter activities.

Animals↗
PubMed1995

Development and migration of olfactory neurones in the nervous system of the neonatal opossum.

The neonatal opossum (Monodelphis domestica) was used to assess how different populations of cells are generated in the olfactory region, and how they migrate along pathways to the central nervous system. Developing nerve cells were immunocytochemically labelled using antisera directed against two specific markers of olfactory receptor neurones: olfactory marker protein (OMP) and the dipeptide carnosine. In new-born opossums both carnosine and OMP are already co-expressed in primary olfactory neurones and in those axons that extend towards the olfactory bulb. Expression of these markers in olfactory receptor neurones during the first postnatal days reflects the advanced developmental state of this system compared to other regions of the central nervous system (such as the cortex and cerebellum), which are highly immature and less developed in comparison with those of new-born rats or mice. A second, distinct population of carnosine/OMP expressing cells was also identified during the first postnatal week. These neurones were present as clusters along the olfactory nerve bundles, on the ventral-medial aspect of the olfactory bulb and in the basal prosencephalon. The distribution of this cell population was compared to another group of well characterized migratory neurones derived from the olfactory placode, which express the decapeptide GnRH (Gonadotropin-releasing hormone, also known as LHRH). GnRH was never co-localized with carnosine/OMP in the same migratory cells. These observations show that distinct cell populations arise from the olfactory placode in the neonatal opossum and that they migrate to colonize the central nervous system by following common pathways.

Animals↗
PubMed1995

Confocal microscopy analysis of NPY and TH immunoreactivities in the hypothalamo-hypophysial system of the frog.

Co-expression of tyrosine hydroxylase (TH) and neuropeptide Y (NPY) in local circuits innervating the hypothalamo-pituitary complex of the green frog, Rana ridibunda, was investigated using simultaneous double immunohistochemical technique, aided by dual-channel confocal laser scanning microscopy. NPY and TH immunoreactivities were observed co-occurring within a discrete neuronal population located in the suprachiasmatic region. In other hypothalamic areas, NPY-immunoreactive (IR) perikarya were generally codistributed, but distinct from TH-IR cells. In the adenohypophysial pars intermedia, the overlap between the two markers was partial, demonstrating the existence of multiple neuronal sources for the inputs to the gland.

Animals↗
PubMed1995

Mapping of neurochemical markers in quail central nervous system: VIP- and SP-like immunoreactivity.

The distribution of cells and fibres containing vasoactive intestinal polypeptide (VIP) and substance P (SP) was investigated in the brain of Japanese quail focussing on the centers involved in reproductive activities. SP-immunoreactive (ir) structures were chiefly present within the ventral telencephalic regions, the periventricular hypothalamus and the dorsal aspects of thalamus. VIP immunopositive structures were rarely associated with recognizable nuclei and they were observed in the organum septi laterale (LSO), the lobus paraolfactorius (LPO), the eminentia mediana (ME), the nucleus striae terminalis (nST) and the area ventralis of Tsai (AVT). SP- and VIP-ir structures were both associated with regions implicated in the control of reproduction. SP was mainly distributed within regions that control male copulatory behavior (the preoptic region, the anterior hypothalamus and the central gray), whereas VIP was prevalently located in the mediobasal hypothalamus that is implicated in the control of female reproductive activities.

Animals↗
PubMed1995

Cell migration from the olfactory placode and the ontogeny of the neuroendocrine compartments.

The olfactory placode and its derivative, the olfactory pit, give rise to several different populations of migrating cells, which contribute to drive the organization of the prosencephalon, but also to form a part of the central neuroendocrine compartments. Some cell types are seemingly transient and can play a role in the establishment of the final connections. The understanding of the mechanisms involved in the migration and differentiation of these cell populations can give an insight on the interplay between peripheral structures and central nervous system and on the mechanisms of commitment, phenotype selection and control for neuroendocrine cells able to selectively "colonize" the brain.

Animals↗
PubMed1994

Glutamate and carnosine in the vestibular system of the frog.

We demonstrate that both glutamate-like and carnosine-like immunoreactivities are present in hair cells and in fibers of the vestibular organ of the frog inner ear. Comparison of the two immunoreactivity patterns indicates that glutamate and carnosine might be colocalized in some hair cells. The presence of glutamate-like immunoreactivity in hair cells is consistent with biochemical and pharmacological data indicating glutamate as the excitatory neurotransmitter in these sensory receptors. There is also evidence that carnosine might have a neuromodulatory function.

Amino Acid Sequence↗
PubMed1994

Multiple sources of the pituitary pars intermedia innervation in amphibians: a DiI retrograde tract-tracing study.

Afferent projections to the pituitary pars intermedia were studied using the DiI tract-tracing technique in two amphibian species, the urodelan Triturus carnifex, and the anuran Rana esculenta. After DiI crystal application into the pituitary intermediate lobe, in both species cells were retrogradely labeled in the preoptic nucleus, in the supra- and retro-chiasmatic hypothalamus and in the brainstem (especially in the area indicated as locus coeruleus). The findings are discussed in relation to data on the neurochemical nature of the innervation of the pars intermedia in amphibians.

Animals↗
PubMed1994

In vitro study of olfactory receptor neurones expressing the dipeptide carnosine.

Olfactory neuroepithelial (OE) cells were dissociated from late stage embryonic mice and analysed for carnosine expression. The yield of carnosine neurones was twice as high when the OE cells were seeded along with the olfactory bulb cells. Carnosine neurones resulted from both in vitro survival and neurogenesis, and were associated with clusters of underlying flat cells immunopositive for keratin. Our results demonstrate that olfactory neurones expressing their neurotransmitter carnosine can be studied in culture, and the close association with keratin-immunopositive basal cells suggests that they are dependent on these cells for survival and/or differentiation.

Animals↗
PubMed1994

Involvement of retinohypothalamic input, suprachiasmatic nucleus, magnocellular nucleus and locus coeruleus in control of melanotrope cells of Xenopus laevis: a retrograde and anterograde tracing study.

The amphibian Xenopus laevis is able to adapt the colour of its skin to the light intensity of the background, by releasing alpha-melanophore-stimulating hormone from the pars intermedia of the hypophysis. In this control various inhibitory (dopamine, gamma-aminobutyric acid, neuropeptide Y, noradrenaline) and stimulatory (thyrotropin-releasing hormone and corticotropin-releasing hormone) neural factors are involved. Dopamine, gamma-aminobutyric acid and neuropeptide Y are present in suprachiasmatic neurons and co-exist in synaptic contacts on the melanotrope cells in the pars intermedia, whereas noradrenaline occurs in the locus coeruleus and noradrenaline-containing fibres innervate the pars intermedia. Thyrotropin-releasing hormone and corticotropin-releasing hormone occur in axon terminals in the pars nervosa. In the present study, the neuronal origins of these factors have been identified using axonal tract tracing. Application of the tracers 1,1'dioctadecyl-3,3,3',3' tetramethyl indocarbocyanine and horseradish peroxidase into the pars intermedia resulted in labelled neurons in two brain areas, which were immunocytochemically identified as the suprachiasmatic nucleus and the locus coeruleus, indicating that these areas are involved in neural inhibition of the melanotrope cells. Thyrotropin-releasing hormone and corticotropin-releasing hormone were demonstrated immunocytochemically in the magnocellular nucleus. This area appeared to be labelled upon tracer application into the pars nervosa. This finding is in line with the idea that corticotropin-releasing hormone and thyrotropin-releasing hormone stimulate melanotrope cell activity after diffusion from the neural lobe to the pars intermedia. After anterograde filling of the optic nerve with horseradish peroxidase, labelled axons were traced up to the suprachiasmatic area where they showed to be in contact with suprachiasmatic neurons. These neurons showed a positive reaction with anti-neuropeptide Y and the same held for staining with anti-tyrosine hydroxylase. It is suggested that a retino-suprachiasmatic pathway is involved in the control of the melanotrope cells during the process of background adaptation.

Adaptation, Physiological↗
PubMed1994

Localization, identification, and action of calcitonin gene-related peptide in the frog adrenal gland.

The localization of calcitonin gene-related peptide (CGRP)-like immunoreactivity in the adrenal gland of the frog, Rana ridibunda, was examined by the indirect immunofluorescence technique. Using an antiserum directed against rat alpha-CGRP, the presence of a network of positive fibers was observed in the adrenal parenchyma. The immunoreactive material has been characterized by HPLC analysis combined with RIA quantification. The elution profile revealed the existence of a single form of CGRP exhibiting the same retention time as synthetic frog CGRP. The possible involvement of CGRP in the regulation of corticosteroid secretion was studied in vitro using a perifusion system for frog adrenal slices. Graded doses of frog CGRP (from 3 x 10(-9) to 3 x 10(-6) M) increased corticosterone and aldosterone secretion in a dose-dependent manner (ED50, 4.1 x 10(-8) M). Several other forms of CGRP, i.e. rat alpha-CGRP and beta-CGRP, and human alpha-CGRP and beta-CGRP, were also capable of enhancing steroid output, but frog CGRP was the most effective stimulator of steroidogenesis. Repeated administration of rat alpha CGRP induced a reproducible stimulation of corticosteroid secretion without any tachyphylaxis. Prolonged infusion of the peptide (3 h) caused a rapid increase in corticosteroid release, followed by a gradual decline of steroid secretion, suggesting the occurrence of a desensitization phenomenon. Rat alpha-CGRP also gave rise to a significant increase in corticosteroid release from acutely dispersed adrenal cells. These results show the presence of CGRP in fibers innervating the frog adrenal gland. The data also demonstrate that synthetic CGRP exerts a direct stimulatory effect on corticosteroid secretion. Taken together, these findings suggest that CGRP, released by nerve fibers in the adrenal tissue, can locally regulate corticosteroid secretion.

Adrenal Cortex Hormones↗
PubMed1994

GnRH neurons and other cell populations migrating from the olfactory neuroepithelium.

Cell migration from the olfactory neuroepithelium to the brain has been widely studied during vertebrate development. Immunocytochemical analysis has revealed that many of the migrating cells contain GnRH (Gonadotropin-Releasing Hormone). The GnRH positive cells migrate from the medial olfactory placode, steam along the nasal septum, cross the basal forebrain and reach the hypothalamic and septal areas from where they control the release of hypophyseal gonadotropic peptides. A peculiar feature of these cells is that they start expressing GnRH during migration. We have analysed the presence of immunoreactivity for peptides typically expressed in olfactory neurones, along the migratory pathway followed by GnRH neurones. We have used polyclonal antibodies raised against carnosine and olfactory marker protein (OMP), and performed double immunolabelling on mouse embryos and on early neonatal Brazilian opossum (Monodelphis domestica) tissues. Beside the GnRH neurones we observed other migrating cells along the pathway traced by olfactory terminal and vomeronasal nerves. Most of these cells co-express carnosine and OMP. The carnosine/OMP migrating cells are detectable in later developmental stages than GnRH neurones. GnRH neurones do not express either OMP or carnosine. By keeping in culture explants of the brain together with the olfactory region from newborn opossums, we have shown that it is possible to obtain the migration of the different populations in vitro. Moreover the GnRH cells are co-distributed, but different from those expressing olfactory markers.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

Presynaptic co-localization of carnosine and glutamate in olfactory neurones.

Olfaction plays a dominant role in modulating behaviour in most vertebrate species and the olfactory bulb is considered a model system for characterizing principles of neural computation. Nevertheless, although the physiology and neurochemistry of the olfactory circuits have been widely studied, the neurotransmitter released by olfactory receptor neurones remains unknown. We now describe the ultrastructural localization of the dipeptide carnosine and the excitatory amino acid glutamate in the glomerular layer of the mouse olfactory bulb. We demonstrate that both carnosine-like and glutamate-like immunoreactivities are selectively co-localized in the olfactory neurone boutons. These observations, taken with the recent findings of glutamate-receptor subunit expression in rodent olfactory bulb, argue compellingly for a role of glutamate in olfactory neurotransmission and suggest a modulatory effect of carnosine.

Animals↗
PubMed1993

In vitro study of catecholamine release from perifused frog adrenal slices.

The adrenal gland of amphibians is composed of a mixed population of adrenochromaffin and corticosteroid-secreting cells. It has previously been shown that chromaffin cells synthesize several bioactive substances (including biogenic amines and neurotransmitters) which may act locally to regulate corticosteroid secretion. In the present report, we have studied the secretory activity of adrenochromaffin cells in Rana ridibunda. Frozen sections of adrenal gland were immunolabeled with antisera against tyrosine hydroxylase or phenylethanolamine-N-methyltransferase. Comparison of homologous fields on consecutive sections indicated that 77% of catecholaminergic cells produce adrenaline. The concentrations of catecholamines were measured by means of high performance liquid chromatography analysis coupled to electrochemical detection. The concentrations of dopamine, noradrenaline, and adrenaline in fresh adrenal tissue were 24 +/- 4, 763 +/- 68, and 1032 +/- 118 ng/mg wet weight, respectively. After a 12-hr perifusion period, the concentration of adrenaline in the tissue was reduced by 62%, whereas noradrenaline only decreased by 22%. The secretion rates of adrenaline and noradrenaline from perifused adrenal slices significantly diminished during the first 7 hr of the experiment and then remained relatively stable for about 10 hr. Exposure of adrenal tissue to a depolarizing concentration of potassium (55 mM) induced an immediate and substantial rise of adrenaline and noradrenaline release and a delayed increase in corticosterone output. Acetylcholine, which stimulates corticosterone secretion from frog adrenocortical cells, induced a slight but not significant increase of adrenaline and noradrenaline release. Similarly, the selective cholinergic agonists muscarine and nicotine did not significantly affect catecholamine release, while muscarine mimicked the stimulatory action of acetylcholine on corticosterone secretion. This study validates the use of the perifusion model to investigate the mechanism of control of catecholamine release from frog adrenochromaffin tissue. The results presented herein indicate that, in contrast to mammals, the secretion of catecholamines from the amphibian adrenal gland is not regulated by cholinergic inputs.

Adrenal Cortex Hormones↗
PubMed1993

Co-occurrence of substance P and enkephalin-like immunoreactivities in the prosencephalon of two amphibians.

We studied the co-occurrence of substance P-(SP) and leucine (or methionine) enkephalin (ENK)-like immunoreactivities in the prosencephalon of two amphibians: Rana esculenta L. (Anura) and Triturus carnifex Laur. (Urodela) using single and double immunohistochemical methods. The two peptides (SP-ENK) were sometimes found colocalized in neuronal cell bodies and processes. Double-labelled neurons were seen in the nucleus accumbens, lateral septal nucleus, preoptic area, dorsal and ventral hypothalamic nuclei of the frog, whereas in the newt co-occurrence was shown in neurons of the primordium pyriform, corpus striatum, pars ventralis thalami, preoptic area and nucleus infundibularis dorsalis and ventralis. Furthermore, in both amphibians, fibres simultaneously showing SP- and ENK-positivity were observed in the amygdaloid complex, preoptic area and in the posterior hypothalamus, around the infundibulum. Co-occurrence studies in comparative neuroanatomy are relevant, as shown in the present paper regarding the similarities of SP-ENK distribution in the striatopallidal and striatonigral systems of tetrapods.

Animals↗