P. de Girolamo

Localization of BDNF expression in the developing brain of zebrafish

The brain‐derived neurotrophic factor (BDNF) gene is expressed in differentiating and post‐mitotic neurons of the zebrafish embryo, where it has been implicated in Huntingtons disease. Little is known, however, about the full complement of neuronal cell types that express BDNF in this important vertebrate model. Here, we further explored the transcriptional profiles during the first week of development using real‐time quantitative polymerase chain reaction (RT‐qPCR) and whole‐mount in situ hybridization (WISH). RT‐qPCR results revealed a high level of maternal contribution followed by a steady increase of zygotic transcription, consistent with the notion of a prominent role of BDNF in neuronal maturation and maintenance. Based on WISH, we demonstrate for the first time that BDNF expression in the developing brain of zebrafish is structure specific. Anatomical criteria and co‐staining with genetic markers (shh, pax2a, emx1, krox20, lhx2b and lhx9) visualized major topological domains of BDNF‐positive cells in the pallium, hypothalamus, posterior tuberculum and optic tectum. Moreover, the relative timing of BDNF transcription in the eye and tectum may illustrate a mechanism for coordinated development of the retinotectal system. Taken together, our results are compatible with a local delivery and early role of BDNF in the developing brain of zebrafish, adding basic knowledge to the study of neurotrophin functions in neural development and disease.

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.

Immunoreactivity to vasoactive intestinal polypeptide (VIP) in the hypothalamus of the domestic fowl, Gallus domesticus

The distribution of VIP-immunoreactive neurons and fibers was detected in the hypothalamus of the chick by immunohistochemistry and light microscopy. A large amount of VIP cellular bodies was localized in the anterior and medial area of the hypothalamus with the highest density of cells in supraoptic, magnocellular preoptic, suprachiasmatic and paraventricular nuclei. Only few VIP-immunoreactive neurons were observed in the caudal section of infundibuli nucleus. A considerable concentration of VIP-positive fibers was also detected in the external layer of the anterior and posterior median eminence. Their presence might have origin both from the neurons of the infundibuli nucleus and from the cells of the paraventricular nucleus. Few VIP-immunoreactive fibers were revealed in the organum vasculosum of the lamina terminalis. These results tend to suggest that VIP may play more than one role in the hypothalamic regions, particularly in the preoptico-hypothalamic area. The presence of this peptide in the median eminence supports even more the hypothesis that it may be released into the portal circulation and transported to the pars distalis of the pituitary gland.

Nerve growth factor in the adult brain of a teleostean model for aging research: Nothobranchius furzeri

Nerve growth factor (NGF) acts on central nervous system neurons, regulating naturally occurring cell death, synaptic connectivity, fiber guidance and dendritic morphology. The dynamically regulated production of NGF beginning in development, extends throughout adult life and aging, exerting numerous roles through a surprising variety of neurons and glial cells. This study analyzes the localization of NGF in the brain of the teleost fish Nothobranchius furzeri, an emerging model for aging research due to its short lifespan. Immunochemical and immunohistochemical experiments were performed by employing an antibody mapping at the N-terminus of the mature chain human origin NGF. Western blot analysis revealed an intense and well defined band of 20 kDa, which corresponds to proNGF of N. furzeri. Immunohistochemistry revealed NGF immunoreactivity (IR) diffused throughout all regions of telencephalon, diencephalon, mesencephalon and rhomboencephalon. It was detected in neurons and in glial cells, the latter mostly lining the mesencephalic and rhomboencephalic ventricles. Particularly in neurons, NGF IR was localized in perikarya and, to a less extent, in fibers. The widespread distribution of proNGF suggests that it might modulate numerous physiological functions in the adult brain of N. furzeri. The present survey constitutes a baseline study to enhance the understanding of the mechanisms underlying the role of NGF during aging processes.

Extrahypothalamic distribution of vasoactive intestinal polypeptide (VIP)-like immunoreactivity in the chicken brain, Gallus domesticus

The distribution of VIP-immunoreactive neurons and fibers was detected in the extrahypothalamic areas of chicken brain by immunohistochemistry and light microscopy VIP-ir perikarya were found in the hippocampus and in the area parahippocampalis; in the area ventralis of Tsai, in the n. interpeduncularis, in the substantia nigra, in the substantia grisea centralis, in the locus coeruleus, in the n. subcoeruleus ventralis and in the n. pontis lateralis. VIP-ir fibers were seen in the lobus parolfactorius and throughout the brainstem mainly arranged in lateral and midsagittal position. This finding was discussed in relation to other studies performed on chicken and/or other avian brain. The distributional pattern of VIP-ergic system in the chicken brain suggests a possible involvement of VIP or VIP-like peptide in several neuroregulatory mechanisms.

Expression and distribution of leptin and its receptors in the digestive tract of DIO (diet-induced obese) zebrafish

The expression and localization of leptin (A and B) and its receptor family in control and diet-induced obese (DIO) adult male zebrafish gut, after 5-weeks overfeeding, administering Artemia nauplii, as fat-rich food, were investigated. Recently, the obese adult zebrafish was considered an experimental model with pathophysiological pathways similar to mammalian obesity. Currently, there are no reports about leptin in fish obesity, or in a state of altered energy balance. By qRT-PCR, leptin A and leptin B expression levels were significantly higher in DIO zebrafish gut than in the control group (CTRL), and the lowest levels of leptin receptor mRNA appeared in DIO zebrafish gut. The presence of leptin and its receptor proteins in the intestinal tract was detected by western blot analysis in both control and DIO zebrafish. By single immunohistochemical staining, leptin and leptin receptor immunoreactive endocrine cells were identified in the intestinal tract either in DIO or control zebrafish. Moreover, leptin immunopositive enteric nervous system elements were observed in both groups. By double immunohistochemical staining, leptin and its receptor were colocalized especially in DIO zebrafish. Thus, our study represents a starting point in the investigation of a possible involvement of leptin in control of energy homeostasis in control and DIO zebrafish.

Neurotrophin-4 in the brain of adult Nothobranchius furzeri

Neurotrophin-4 (NT-4) is a member of the well-known family of neurotrophins that regulate the development of neuronal networks by participating in neuronal survival and differentiation, the growth of neuronal processes, synaptic development and plasticity, as well as myelination. NT-4 interacts with two distinct receptors: TrkB, high affinity receptor and p75 low-affinity neurotrophin receptor (p75(NTR)). In the present survey, we identified the gene encoding NT-4 in the teleost Nothobranchius furzeri, a model species for aging research. The identified gene shows a similarity of about 72% with medaka, the closest related species. The neuroanatomical localization of NT-4 mRNA is obtained by using an LNA probe. NT-4 mRNA expression is observed in neurons and glial cells of the forebrain and hindbrain, with very low signal found in the midbrain. This survey confirms that NT-4 is expressed in the brain of N. furzeri during adulthood, suggesting that it could also be implicated in the maintenance and regulation of neuronal functions.

RET receptor in the gut of developing cat

RET receptor is a transmembrane protein which, together with the glial-cell-line derived neurotrophic factor family receptors alpha, forms a receptor complex upon activation by the glial-cell-line-derived neurotrophic ligands (GFLs). RET signaling is crucial for: (a) development of the enteric nervous system and kidney; (b) development of sympathetic, parasympathetic, motor, and sensory neurons; (c) postnatal maintenance of dopaminergic neurons; (d) spermatogenesis. In humans, RET mutations cause the Hirschsprungs disease, characterized by megacolon aganglionosis, and different types of cancer, the multiple endocrine neoplasia type 2A and type 2B and familial medullary thyroid. In the earliest aged cat embryos studied (stage 9 according to Knopse), RET immunoreactivity (IR) was observed in few cells detected in bilateral rows extending latero-ventrally to the neural tube and dorso-laterally to the foregut. In the successive aged group (stage 11), RET IR was observed in few single or grouped epithelial cells of the anterior gut and in small clustered cells scattered in the mesenchyme around the anterior gut. From stage 14-22 (the last stage 22 includes foetuses around the birth), RET IR was seen in neurons and fibers of the enteric nervous system. The appearance and intensification of RET-IR in the gut occurred with cranio/caudal and external/internal directions during the development. These results, thus, suggest the involvement of GFLs in the neuroblast migration, proliferation and differentiation. For a short period of development, these molecules might also act on some cells of the epithelium.

GDNF and GFRα co-receptor family in the developing feline gut.

Glial cell-line derived neurotrophic factor (GDNF) and the GFRα co-receptors play a role in the developing enteric nervous system. The co-receptors elicit their action by binding receptor tyrosine kinase RET. This immunohistochemical study reports the presence of GDNF and its specific co-receptor GFRα1 in the cat gastrointestinal apparatus during development, from stage 9 to 22. At stage 9 and 11, immunoreactivity (IR) to GDNF was observed in the cells of mesenchyme of the anterior gut. From stage 14 to 22, GDNF IR was detected in nervous plexuses; moreover, GDNF and GFRα1 IR appeared localized in gastrointestinal endocrine cells. The presence of GDNF in the enteric nervous system and in the endocrine cells suggests an involvement of this neurotrophic factor in the gastrointestinal development. Moreover, the presence of the co-receptor GFRα1 in endocrine cells and its absence in the enteric nervous system seems to indicate a different mode of transduction of GDNF signal. GFRα2 and GFRα3 co-receptors were not detected.

The Innervation of the Fetal Buffalo Tongue

, 1998).Since the lingual papillae of the domestic ruminant show an extreme variety of shape, webelieved it interesting to extend the study to innervation of the lingual papillae of the waterbuffalo, analyzing the morphological, structural, and immunohistochemical characteristicsof the gustatory papillae.MATERIALS AND METHODSThe study was performed on 25 water buffalo fetuses of both sexes, from 5 to 60cm inlength, and originating from a local slaughter-house. The fetus length, from the sinciput tothe tail root, was utilized as a development index. The fetuses were subdivided into fivegroups in relation to their length (5–7cm; 10–15cm; 20–25cm; 30–35cm; 40–60cm). Foreach group samples were prepared for scanning electron microscopy (SEM), transmissionelectron microscopy (TEM), light microscopy (LM), and the simple immunohistochemicalmethod.The TEM samples were treated by the usual techniques.