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G Tagliafierro

Publications and source records attributed to G Tagliafierro.

At least 19 recordsLinked to original sources
PubMed2003

Involvement of acetyl choline in settlement of Balanus amphitrite.

The aim of the present study was to investigate the presence and distribution of cholinergic molecules in Balanus amphitrite cyprids and their possible involvement in settlement and adhesion. Acetylcholinesterase (AChE, the lythic enzyme of acetylcholine) activity was detected, for the first time, by biochemical and histoenzymological methods, in the thoracic muscles, gut wall and cement gland. The immunodetection of choline acetyltransferase-like (ChAT) molecules in the same area and in the neuropil of the central nervous system suggests the presence of a cholinergic innervation, and the involvement of acetylcholine in muscular contraction and cement gland exocytosis. The binding of FITC-conjugate alpha-bungarotoxin in the cement gland cells confirms the latter hypothesis. Acetylcholine involvement in the settlement process was also investigated by laboratory tests employing cholinergic antagonists and agonists. An increase of available acetylcholine due to the partial inhibition of AChE activity produced an increase in cyprid settlement. The data presented support the hypothesis that acetylcholine has a neurotransmitter/neuromodulator role in settlement and adhesion of barnacle cyprids.

Acetylcholine↗
PubMed2002

Synchronized onset of nuclear and cell surface modifications in U937 cells during apoptosis.

In this study we investigated the relationship between nuclear and cell surface modifications (i.e. blebbing, phosphatidylserine [PS] and sugar residues exposure) in a monocytic cell line, U937, during apoptosis induced by oxidative stress (1 mM H2O2) or inhibition of protein synthesis (10 microg/ml puromycin). Dying cells were simultaneously observed for nuclear modifications, presence of superficial blebs and plasma membrane alterations. Morphological analysis performed by conventional fluorescence microscopy, or by transmission and scanning electron microscopy showed that the courses of nuclear and membrane alterations occured concomitantly, but the phenotype was dependent on the stage of the apoptotic process and the type of apoptogenic inducer used. The progression of apoptosis in U937 cells beyond early stages resulted in the extensive formation of blebs which concomitantly lost some typical markers of apoptosis, such as PS and sugar residues. Therefore, the modality by which the nucleus condenses, or the amount and the pattern of distribution of PS on the cell surface were, for each cell line, strictly related to the apoptogenic inducer. The morphological data reported in the present paper should lead to a more precise quantification of apoptosis by improving the detection of apoptotic cells in vivo (i.e. in tissue, organs), which is a crucial point in the evaluation of efficiency of antiproliferative drugs, such as antiblastic or immunosuppressive compounds.

Apoptosis↗
PubMed2002

Microgravity-induced apoptosis in cultured glial cells.

Apoptosis is a form of naturally occurring cell death that plays fundamental roles during embryonic developement. In adults, it neatly disposes of cells damaged by injuries provoked by external causes such as UV radiation, ionisation and heat shock. Alteration of the gravity vector may be one of the external apoptosis inducers. Neurophysiological impairment signs were seen during space flights in astronauts, but very few studies were carried out on the nervous system and none at the cellular level. In this study, we submitted cultured C6 glioma cells to microgravity (0xg) of varying duration, obtained by clinorotation in a Fokker three-dimensional clinostat for 15 min, 30 min, 1h, 20h or 32h. After 30 min at 0xg, numerous nuclei underwent the classical morphological alterations (chromatin condensation, nuclear fragmentation, apoptotic bodies) that lead to the programmed cell death. After 30 min at 0xg, immunostaining for the enzyme caspase-7 was present in the cytoplasm of many cells concurrently with DNA fragmentation identified by the TUNEL method. At 32h, the number of apoptotic nuclei was much reduced indicating the ability of glial cells to adapt to altered gravity.

Animals↗
PubMed2002

Microgravity-induced programmed cell death in astrocytes.

Cultured astrocytes were submitted to simulated microgravity using a Fokker clinostat under continuous rotation (60 rpm) for 15', 30', 1h, 20h and 32h. Samples processing included (i) nuclear stainings using Propidium Iodide and 4,6-diamidino-2-phenilindole, dihydro chloride, (ii) immunohistochemical identification of Caspase-7, (iii) identification of DNA fragmentation using the terminal dUTP nick end labelling and (iv) Scanning Electron Microscope analysis. After 30' at simulated microgravity the glial cells showed morphological evidence of apoptosis: cell shrinkage, chromatin condensation, nuclear blebs and fragmentation. The enzyme caspase-7 was present and DNA fragmentation was evident. After 32h the density of the cell population was much lower than that observed in controls.

Journal Article↗
PubMed2001

Organization of vasoactive intestinal peptide-like immunoreactive system in the brain, olfactory organ and retina of the zebrafish, Danio rerio, during development.

The localization of vasoactive intestinal peptide (VIP)-like immunoreactivity was investigated in the brain, olfactory system and retina of the zebrafish, Danio rerio, during development and in juvenile specimens, by using the indirect immunofluorescence and the peroxidase-antiperoxidase methods. In 24 h post fertilization (hpf) embryos, VIP-like immunoreactive cells were present in the olfactory pit, the retina, and several regions of the brain, including the dorsal telencephalon, the diencephalon, the tegmentum of the mesencephalon, the caudal rhombencephalon and the anterior pituitary. In 48 hpf embryos, additional VIP-like immunoreactive cell bodies were found in the ventral telencephalon, whereas in the diencephalon VIP-like immunopositive cells were more concentrated within the ventro-caudal hypothalamus. During the 7 day larval period, a dense plexus of VIP-like immunoreactive fibers first appeared in the olfactory bulbs. In 15-day-old larvae, two new groups of positive cells were observed in the periventricular preoptic nucleus and in the dorsal rhombencephalon. In 1 month/2 months old animals, VIP-like immunoreactive elements were confined to the olfactory organ, the olfactory bulbs, the periventricular preoptic nucleus and the pituitary, pars distalis. At 3 months stage, a large number of cells was observed in the periventricular preoptic nucleus. Western immunoblot analysis confirmed that VIP-like peptides, with molecular weight similar to that of synthetic VIP, are present early during the development of zebrafish. These results show that VIP-like immunoreactive structures appear early during ontogeny both in the olfactory pit, retina and brain. Transient expression of positive cells was found in the retina, telencephalon, diencephalon and brainstem. The location of VIP-like immunoreactivity indicates that, during development, VIP could be involved in several neuromodulatory functions, including the processing of visual and olfactory informations, as well as growth or survival promotion activities. The presence of VIP-like immunopositive cells in the pituitary, pars distalis, suggest that, during development, VIP may influence the secretion of pituitary hormones.

Aging↗
PubMed2001

Three-dimensional computer-aided reconstruction of FMRFamide immunopositive neuron distribution in the ventral ganglion of the barnacle Balanus amphitrite (Cirripedia, Crustacea).

We have implemented a simple program to solve three of the problems related to 3D reconstruction (3D-Rec) of soft tissues: alignment of sections, distortions, and estimation of the spatial position of elements of interest inside the tissues. As a model, we chose the distribution of FMRFamide-like immunopositive neurons in the ventral ganglion of the barnacle Balanus amphitrite collected during different seasonal periods. Images of immunostained sections were acquired by means of a CCD-camera-equipped microscope and a PC and the reference points were taken inside the sections. The FMRFamide-like immunopositive neurons detected in the barnacle ventral ganglion were grouped into four different classes according to size, shape and staining intensity. More numerous FMRFamide-like immunopositive neurons were detected in the autumn-collected barnacle than in the summer counterpart. The two 3D reconstructions obtained from transverse and longitudinal ventral ganglion sections were efficaciously compared after 90 degrees rotation of one of them. Comparison of these two 3D-Rec suggests the presence of at least two groups of FMRFamide-like immunopositive neurons that are seasonally-related and probably involved in reproduction.

Animals↗
PubMed2001

Simulated microgravity induces alteration in the central nervous system.

To investigate whether the signs of neurophysiological impairment observed in flight may be traced back to cytomorphology, we undertook a ground-based study focusing upon the architecture of cultured glial cells under simulated microgravity obtained by three-dimensional clinorotation.

Actin Cytoskeleton↗
PubMed1999

Macronuclear chromatin structure dynamics in Colpoda inflata (Protista, Ciliophora) resting encystment.

The chromatin structure dynamics of the Colpoda inflata macronucleus have been investigated in relation to its functional condition, concerning chromatin body extrusion regulating activity. Samples of 2- and 25-day-old resting cysts derived from a standard culture, and of 1-year-old resting cysts derived from a senescent culture, were examined by means of histogram analysis performed on acquired optical microscopy images. Three groups of histograms were detected in each sample. Histogram classification, clustering and matching were assessed in order to obtain the mean histogram of each group. Comparative analysis of the mean histogram showed a similarity in the grey level range of 25-day- and 1-year-old cysts, unlike the wider grey level range found in 2-day-old cysts. Moreover, the respective mean histograms of the three cyst samples appeared rather similar in shape. All this implies that macronuclear chromatin structural features of 1-year-old cysts are common to both cyst standard cultures. The evaluation of the acquired images and their respective histograms evidenced a dynamic state of the macronuclear chromatin, appearing differently condensed in relation to the chromatin body extrusion regulating activity of the macronucleus. The coexistence of a chromatin-decondensed macronucleus with a pycnotic extrusion body suggests that chromatin unable to decondense, thus inactive, is extruded. This finding, along with the presence of chromatin structural features common to standard and senescent cyst populations, supports the occurrence of 'rejuvenated' cell lines from 1-year-old encysted senescent cells, a phenomenon which could be a result of accomplished macronuclear renewal.

Animals↗
PubMed1997

Islets and diffuse endocrine component in the pancreas of three red frogs species: relationships between endocrine and exocrine tissue.

The endocrine pancreas of three red frogs was studied immunohistochemically. It consisted of islets and diffuse endocrine cells. The islets showed a mammalian-like arrangement with a central core of B cells and a peripheral mantle of A/PP cells. A few D and VIP cells were also present. Several regulatory peptides were co-localized in the same endocrine cells by consecutive sections and double-labeling studies. The A/PP cells were formed by subpopulations of cells showing various types of immunoreactivity and varying degrees of immunolabeling. Generally, glucagon/pancreatic polypeptide, glucagon/pancreatic polypeptide/peptide tyrosine tyrosine and glucagon/pancreatic polypeptide/neuropeptide tyrosine immunoreactivities were present in the islets and in the endocrine cells scattered throughout the exocrine parenchyma (the diffuse component). Some specimens, mainly belonging to Rana dalmatina, showed evident periinsular halos around the islets. The diffuse component was abundant, and mainly contained A/PP cells. It formed a net across the exocrine parenchyma; its interrelationship with the latter might occur by a paracrine mechanism.

Animals↗
PubMed1997

The gastro-enteric-pancreatic neuroendocrine system in two reptilian species: Chalcides chalcides and Zoonosaurus madascariensis (Sauridae).

The presence, localization and distribution of some regulatory peptides and serotonin were investigated by single and double immunohistochemical methods in the digestive system of two reptiles, Chalcides chalcides and Zonosaurus madascariensis. Both immunoreactive (IR) cells and nerve elements were demonstrated, showing different distributions according to the antisera tested. Similar results were observed in the two saurian species. Chromogranin-SPI-, serotonin-, somatostatin14-, pancreatic polypeptide (PP)-, and gastrin-IR cells were present along the gut epithelium. Cholecystokinin (CCK)-, and insulin-IR cells seemed to be more concentrated in the intestinal portion, while very few glucagon containing cells were observed. Bombesin-IR cells were found in the stomach and they constituted the only endocrine cells of the closed type in the gut. Vasoactive intestinal polypeptide (VIP)-, insulin-, and bombesin-IR nerve terminals were also seen. In the pancreatic duodenal portion of Z madascariensis, the insulin-, glucagon-, PP-, and somatostatin 14-IR cells were present as single elements or grouped in endocrine islets showing a typical topographical distribution. By double immunohistochemical techniques, chromogranin-SPI was found co-localized with the serotonin- and somatostatin-immunoreactivity, but CCK-IR cells were always negative to chromogranin-SP1 antiserum. The present work demonstrates that the chromogranin antiserum is not useful for identifying all the gut endocrine cell types; furthermore, the presence of insulin-immunoreactivity in the endocrine cells is confirmed, and, for the first time, insulin-immunoreactivity is shown in reptilian gut nerve fibres.

Animals↗
PubMed1996

Immunocytochemical detection of islet hormones in the digestive system of Protopterus annectens.

The presence, distribution, and interrelationships of the four typical pancreatic islet hormones were investigated in the digestive system of Protopterus annectens by single and double immunohistochemical methods. Insulin-, glucagon-, and somatostatin-immunoreactive (IR) elements were detected in both the pancreas and the gut. Pancreatic polypeptide (PP)-IR endocrine cells were always present in the gut, but were only present in the pancreas of a few specimens. Some of the latter cells also seemed to react with glucagon antiserum. In the pancreas the immunopositive cells were organized into islets of different sizes, and their organizations were studied by the double immunohistochemical techniques. In the few large islets insulin-IR cells were present in the central zone, glucagon- and PP-IR cells at the periphery, and somatostatin-IR cells intermingled with both the peripheral and the central endocrine cells. In the smaller islets, the number and the staining intensity of glucagon- and PP-IR endocrine cells varied markedly. In the gut, insulin-, somatostatin-, and PP-IR cells were of the open type; glucagon-containing cells were very few and had no luminal contact. They were differently distributed along the intestinal epithelium. Somatostatin-IR nerve fibers and somatostatin-IR neuron cell bodies were also observed in the intestinal wall. The organization of pancreatic endocrine cells in P. annectens is similar to that observed in the majority of teleosts even if a different topographical association can be found. Furthermore, islets of different sizes seem to display a different metabolic turnover, and the detection of pancreatic PP-immunoreactivity varied according to the specimens utilized. In the intestinal portion insulin-IR cells, in addition to PP-, glucagon- and somatostatin-IR cells are present: this suggests that intestinal insulin-like immunoreactivity may be more widespread than previously supposed.

Animals↗
PubMed1995

Cell composition and co-stored peptides in the endocrine pancreas of Rana arvalis.

The endocrine pancreas of Rana arvalis studied by immunochemistry consisted of islets and diffuse endocrine cells. The islets showed a mammalian-like arrangement with a central core of B cells and a peripheral mantle of A/PP cells. A few D and vasoactive intestinal polypeptide cells were also present. Consecutive sections or double-labeling studies allowed us to detect several regulatory peptides colocalized in the same endocrine cells. The so-called A/PP cells contained subpopulations of cells showing various types of immunoreactivity and varying degrees of immunolabeling. Generally, glucagon/pancreatic polypeptide, glucagon/pancreatic polypeptide/phe-met-arg-phe-amide, glucagon/pancreatic polypeptide/peptide tyrosine tyrosine, glucagon/pancreatic polypeptide/peptide tyrosine tyrosine/phe-met-arg-phe-amide immunoreactivities were present in the islets, while peptide tyrosine tyrosine/neuropeptide tyrosine colocalization was also found in the parenchyma.

Amino Acid Sequence↗
PubMed1995

Cytokeratin type distribution in the skin and gill epithelia of the Indian freshwater catfish, Heteropneustes fossilis as detected by immunohistochemistry.

Based on the general cross-reactivity of the cytokeratins in vertebrates, we describe the immunoreactivity for some mammalian cytokeratins of both the epidermis and gill of H. fossilis. The following monoclonal antibodies, commercially supplied, were tested: K8.13, KL1, AE1 and AE3, which have a wide spectrum of specificity, and LDS-68, M 20, K8.60, KS-B17.2, K4.62, which are more narrowly specific. The reaction of the epithelial cells of the skin to K8.13 was negative in the basal layer, weakly positive in the layers above but strongly positive in some cells of the superficial layer. KL1 was negative in the basal layer, positive in the outer layers. AE1 was strongly positive in the basal layer, negative in the superficial cells. AE3 gave a general but weak reaction in the epithelial cells. K8.60 was negative for the epithelial cells, but reacted positively in the club cells. Club cells also reacted to K8.13 and AE1, and were strongly positive to KL1 and AE3. Goblet mucous cells were negative to all antibodies tested. In the gills, K8.13 labelled cells of both filament and lamellar epithelia. The reaction to AE3 was similar but less intense. KL1 was positive in the basal layer of the filament epithelium but not elsewhere, and K8.60 was negative throughout. AE1 and KS- B17.2 specifically labelled small cells scattered in the filament and lamellar epithelia, which are tentatively identified as neuroendocrine cells.

Animals↗
PubMed1993

Ectopic expression of bioactive peptides and serotonin in the sacciform gland cells of teleost skin.

Immunohistochemical methods identified serotonin, and the peptides bombesin and caerulein, in the skin of the teleosts Lepadogaster candollei and Mastacembelus erytrotaenia. In both species, the secretory content of epidermal sacciform cells reacted positively for all three substances. These results are compared with studies on the skin glands of amphibians, which also contain multiple active compounds, and on various neuroendocrine cells of fish. The precise functions of the secretions are not known.

Animals↗
PubMed1993

Immunoreactive endocrine cells and nerve elements in the gut of the Italian cave salamander.

The presence and distribution of eleven different types of immunoreactive endocrine cells and nine types of immunoreactive nerve elements were immunohistochemically identified in the gut and pancreas of the italian cave salamander, Hydromantes ambrosii. The majority of gastrointestinal endocrine cells were of open-type, often presenting basal cytoplasmic processes. Gastrin- and substance P-immunoreactive cells in the fundus and bombesin-immunoreactive cells in the intestinal portion were instead of closed type. Immunoreactive nerve fibres were particularly numerous in the muscular layers and blood vessel wall; bombesin- and substance P-immunoreactive nerve fibres were also abundant beneath gastro-intestinal epithelium. Besides substance P-, caerulein- and cholecystokinin-immunoreactive nerve fibres, all the other immunopositive nerve fibres seemed to be of intrinsic types. By the use of four different gastrin/cholecystokinin antisera three variously distributed subpopulations of endocrine cells and nerve elements were detected. Most of the pancreatic endocrine cells were organised in chord-like islets, polarized in the direction of blood vessels. A sparse network of bombesin-immunoreactive fibres was also found in the pancreas. The distribution of bombesin- and of the gastrin/cholecystokinin-immunoreactive material in the stomach and the presence of closed type endocrine cells indicate a more evoluted organization of the gastroenteropancreatic neuroendocrine system thus confirming the position of Hydromantes ambrosii among the higher urodeles.

Animals↗
PubMed1992

Ultrastructure and immunocytochemistry of the neuroepithelial bodies in the lung of the tiger salamander, Ambystoma tigrinum (Urodela, Amphibia).

Light and electron microscopy of the lungs of Ambystoma tigrinum (Urodela) revealed a relatively complex pattern of the neuroendocrine (NE) cells. In the apical parts of smaller septa single NE cells not associated with nerve fibres were covered and surrounded by pneumocytes. The larger septa possessed small areas of ciliated epithelium, in which the NE cells were grouped in a form of neuroepithelial bodies (NEB) consisting of 3-5 cells and covered by goblet cells. NE cells possessed a large nucleus with patches of condensed chromatin, clear cytoplasm, and membrane-bound vesicles of variable morphology and size, containing an electron dense interior surrounded by a lucent space. The size of these dense core vesicles (DCV) ranged from 70-140 nm, while rarely the larger ones exhibited a diameter of 300-600 nm. In some NEB a second type of NE cells was observed for the first time in an amphibian species: these cells communicated with the air space and exhibited on their surface microvilli and a single modified cilium with a 8 + 1 microtubule arrangement. Their cytoplasm contained two types of DCV: dense core granules with a diameter of 140-260 nm and vesicles 320-700 nm in diameter with a moderately electron dense interior. The NEB were associated with intracorpuscular, sensory nerve terminals morphologically afferent and efferent. By immunocytochemistry, the NE cells revealed the presence of serotonin, met-enkephalin, and leu-enkephalin. A paracrine and chemoreceptor role is proposed for NEB of Ambystoma tigrinum.

Ambystoma↗
PubMed1990

Serotonin-like immunoreactivity in the epidermal club cells of teleost fishes.

The present immunocytochemical study concerns the distribution of serotonin in the epidermis of three species of teleost fish. Serotonin-like immunoreactivity was found in the club cells of Heteropneustes fossilis and Carapus acus but not in those from the sea eel Conger conger. This study is the first immunocytochemical identification of serotonin in the club cells of teleost epidermis. By comparing data from the literature (Zaccone et al. 1986, 1987, 1988) regarding the occurrence of serotonin and GRP/bombesin in the exocrine sacciform gland cells of piscine skin, it is worthy mentioning here that the serotonin contained in the club cells of the species studied may have the ability to affect the pheromonal or other possible functions of these cells. The presence of serotonin in these systems has been correlated with the capacity of the exocrine glands of fish skin to secrete, ectopically, amine messengers in contrast to those produced eutopically i.e. in the neuron-paraneuron system in some vertebrates (Fujita et al. 1988).

Animals↗