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D Miceli

Publications and source records attributed to D Miceli.

At least 19 recordsLinked to original sources

Retinal and cortical afferents to the dorsal lateral geniculate nucleus of the turtle, Emys orbicularis: a combined axonal tracing, glutamate, and GABA immunocytochemical electron microscopic study.

The dorsal lateral geniculate nucleus (GLd) of the turtle Emys orbicularis has been analyzed with axonal tracing methods and immunocytochemical techniques for glutamate (GLU) and gamma-aminobutyric acid (GABA), in combination with a quantitative study of the morphologic characteristics, distribution, and synaptology of the retinofugal and corticofugal terminals. Ultrastructural observations show that the vast majority of retinal terminals (Rtr) have clear, rounded synaptic vesicles and account for 16% of all profiles containing synaptic vesicles (PCSV). Their synaptic index (0.5) is low, and they make three times more contacts with the dendrites of projection cells than with those of interneurons. A low proportion of retinal terminals of a second category contain pleomorphic synaptic vesicles and are highly GABA immunoreactive. Axon terminals, unlabeled after intraocular injection of the tracer (SR), smaller in size and with more rounded clear synaptic vesicles, longer synaptic differentiations, and higher synaptic index than Rtr terminals, account for 19.7% of all PCSV and make asymmetric synaptic contacts with large dendrites of projection cells and less with the dendrites of interneurons. Some SR have been unambiguously identified as corticofugal terminals (Cg), either after cortical injection of the tracer (16%) or cortical lesion (37%). Retinal and Cg/SR terminals are spatially segregated within the GLd. Both are highly GLU immunoreactive, with the highest density of labeling over synaptic vesicles, suggesting that these terminals may use GLU as neurotransmitter. The level of GLU immunoreactivity of GABA-positive profiles is half that of Rtr and Cg/SR terminals and is greatest over mitochondria, possibly reflecting the 'metabolic' pool of GLU that serves as a precursor in the formation of GABA.

Afferent Pathways

Differential glutamate immunoreactivity in glial cells of the retino-recipient layer of the viper optic tectum following retinal ablation. A quantitative EM immunogold study.

In normal conditions, retino-tectal terminals are densely glutamate-immunoreactive. During the degenerative process of these terminals, a significant increase of glutamate immunoreactivity has been exclusively observed in microglial cells. It is suggested that this phenomenon is consecutive to the synthesis of glutamate by these cells after their activation by degenerating optic terminals.

Animals

Enrichment of glutamate-like immunoreactivity in the retinotectal terminals of the viper Vipera aspis: an electron microscope quantitative immunogold study.

A post-embedding immunogold study was carried out to estimate the immunoreactivity to glutamate in retinal terminals, P axon terminals and dendrites containing synaptic vesicles in the superficial layers of the optic tectum of Vipera. Retinal terminals, identified following either intraocular injection of tritiated proline, horseradish peroxidase (HRP) or short-term survivals after retinal ablation, were observed to be highly glutamate-immunoreactive. A detailed quantitative analysis showed that about 50% of glutamate immunoreactivity was localized over the synaptic vesicles, 35.8% over mitochondria and 14.2% over the axoplasmic matrix. The close association of immunoreactivity with the synaptic vesicles could indicate that Vipera retino-tectal terminals may use glutamate as their neurotransmitter. P axon terminals and dendrites containing synaptic vesicles, strongly gamma-aminobutyric (GABA)-immunoreactive, were shown to be also moderately glutamate-immunoreactive, but two to three times less than retinal terminals. Moreover, in P axon terminals, the glutamate immunoreactivity was denser over mitochondria than over synaptic vesicles, possibly reflecting the 'metabolic' pool of glutamate, which serves as a precursor in the formation of GABA.

Animals

Ultrastructural and phenotypic characterization of CABA I, a new human ovarian cancer cell line.

We have established an ovarian cancer cell line (CABA I) from ascitic fluid obtained from a patient with papillary adenocarcinoma of the ovary prior to drug treatment. The epithelial origin of the cell line was confirmed by morphology and by immunofluorescence analysis using anticytokeratin antibodies. Ultrastructural analysis revealed a very irregular membrane surface and a clear cytoplasm rich in electron-lucent vesicles. CABA I cells grow rapidly in culture (doubling time 18 h) in an anchorage-independent manner. Exogenously added beta-estradiol and epidermal growth factor (EGF) treatments did not influence cell growth rate. FACS analysis to determine the phenotypic profile of tumor-associated antigen, membrane receptor, and adhesion molecule expression indicated that the cell line was positive for different members of the c-erbB family, for alpha 6 and beta 1 integrin receptors, and intensively positive for HLA class I antigens and the folate receptor. Molecular characterization revealed no mutations for c-myc and c-k-ras genes, but did detect an exon 5 mutation in the p53 gene. CABA I cells grew poorly as heterotransplants in nude mice, and tumors showed long latency periods. Because early (15-20) and late (55-60) passage cells maintain the same growth and phenotypic characteristics, the CABA I cell line might provide a good in vitro model system to investigate the cellular and molecular events involved in ovarian carcinogenesis.

Animals

Involvement of the 12-lipoxygenase pathway of arachidonic acid metabolism in homosynaptic long-term depression of the rat hippocampus.

Low-frequency stimulation is associated with long-term depression (LTD) of synaptic efficacy in various brain structures. Like long-term potentiation (LTP), homosynaptic LTD in area CA1 of the hippocampus appears to require NMDA receptor activation, changes in postsynaptic calcium concentration and phospholipase A2 (PLA2) activation. Arachidonic acid (AA) is released after the activation of calcium-dependent phospholipases and free AA is rapidly metabolized to a family of bioactive products (the eicosanoids) which are thought to be both intracellular and extracellular messengers. In the present study, we investigated the involvement of the cyclooxygenase and lipoxygenase pathways of AA metabolism in the formation of homosynaptic LTD in the rat hippocampus. Stimulation at 1 Hz for 15 min was used to produce homosynaptic depression in area CA1 of hippocampal slices. LTD induction was partially blocked by bromophenacyl bromide (50-100 microM), a selective PLA2 inhibitor, and by the a nonselective lipoxygenase inhibitor nordihydroguaiaretic acid (NDGA; 100 microM). In contrast, the specific cyclooxygenase blocker indomethacin (100 microM) did not significantly reduce hippocampal LTD. Since NDGA interferes with LTD formation, we examined whether specific inhibitors of 5- and 12-lipoxygenases were capable of blocking LTD expression. The 12-lipoxygenase inhibitor baicalein at a concentration of 50 microM reduced LTP formation when given in the bath, an effect that was less pronounced with the 5-lipoxygenase inhibitor AA-861. These data suggest that the activation of endogenous PLA2 and the formation of 12-lipoxygenase metabolites of AA may be important factors controlling the expression of hippocampal LTD.

Acetophenones

Distribution of serotonin-immunoreactivity in the brain of the pigeon (Columba livia).

The distribution of serotonin (5-HT)-containing perikarya, fibers and terminals in the brain of the pigeon (Columba livia) was investigated, using immunohistochemical and immunofluorescence methods combined with retrograde axonal transport. Twenty-one different groups of 5-HT immunoreactive (IR) cells were identified, 2 of which were localized at the hypothalamic level (periventricular organ, infundibular recess) and 19 at the tegmental-mesencephalic and rhombencephalic levels. Ten of the cell groups were situated within the region of the midline from the isthmic to the posterior rhombencephalic level and constituted the raphe system (nucleus annularis, decussatio brachium conjunctivum, area ventralis, external border of the nucleus interpeduncularis, zona peri-nervus oculomotorius, zona perifasciculus longitudinalis medialis, zona inter-flm, nucleus linearis caudalis, nucleus raphe superior pars ventralis, nucleus raphe inferior). The 9 other cell populations belonged to the lateral group and extended from the posterior mesencephalic tegmentum to the caudal rhombencephalon [formatio reticularis mesencephali, nucleus ventrolateralis tegmenti, ectopic area (Ec) of the nucleus isthmo-opticus (NIO), nucleus subceruleus, nucleus ceruleus, nucleus reticularis pontis caudalis, nucleus vestibularis medialis, nucleus reticularis parvocellularis and nucleus reticularis magnocellularis]. Combining the retrograde axonal transport of rhodamine beta-isothiocyanate (RITC) after intraocular injection and immunohistofluorescence (fluoresceine isothiocyanate: FITC/5-HT) showed the centrifugal neurons (NIO, Ec) to be immunonegative. Serotonin-IR fibers and terminals were found to be very broadly distributed within the brain and were particularly prominent in several structures of the telencephalon (archistriatum pars dorsalis, nucleus taeniae, area parahippocampalis, septum), diencephalon (nuclei preopticus medianus, magnocellularis, nucleus geniculatus lateralis pars ventralis, nucleus triangularis, nucleus pretectalis), mesencephalon-rhombencephalon (superficial layers of the optic tectum, nucleus ectomamillaris, nucleus isthmo-opticus and in most of the cranial nerve nuclei). Comparing the present results with those of previous studies in birds suggests some major serotonin-containing pathways in the avian brain and clarifies the possible origin of the serotonin innervation of some parts of the brain. Moreover, comparing our results in birds with those obtained in other vertebrate species shows that the organization of the serotoninergic system in many regions of the avian brain is much like that found in reptiles and mammals.

Animals

Retinal and non-retinal inputs upon retinopetal RMA neurons in the lamprey: a light and electron microscopic study combining HRP axonal tracing and GABA immunocytochemistry.

A light and electron microscopic study, combining HRP axonal tracing or degeneration and GABA immunocytochemistry, was performed in the lamprey Lampetra fluviatilis in order to analyze retinal and non-retinal inputs upon the retinopetal neurons localized in the reticular mesencephalic area (RMA). The iontophoretic deposit of HRP onto the central stump of the cut optic nerve produced a dense anterograde labeling in the retino-recipient strata marginale and cellular externum of the optic tectum as well as the retrograde labeling of retinopetal neurons in the mesencephalic tegmentum. The large ascending proximal dendrites of the retinopetal neurons constituted a distinct bundle coursing first dorso-laterally in the dorsal mesencephalic tegmentum, and then dorso-medially in the strata fibrosum centrale and cellulare et fibrosum internum of the optic tectum before their distal portions penetrated the retino-recipient tectal layers. The distribution of GABA immunoreactivity was also investigated in the tectal layers and dorsal mesencephalic tegmentum with both pre- and post-embedding methods. The retinal terminals, identified either following HRP iontophoresis in the optic nerve or in early phases of degeneration after short-term survivals following retinal lesion, contained rounded-shaped synaptic vesicles and were always GABA immunonegative. They established asymmetrical synaptic contacts on the distal dendrites of RMA neurons and represented 11.4% of all terminals contacting such neurons (15% of these neurons were GABA immunopositive). The dense extra-retinal input upon the retinopetal RMA neurons was composed of five types of axon terminal profiles, either GABA-immunopositive or -immunonegative. Considering the different cytochemical types of axon terminals contacting RMA neurons, as well as the characteristics of the retinal targets of these neurons, we suggest that, globally, the effects of RMA neurons upon the retina are mainly inhibitory.

Afferent Pathways

Androgen and estrogen receptors are present in primary cultures of human synovial macrophages.

Macrophages, as antigen-processing and -presenting cells to T lymphocytes, play a key role in the immune system and are suspected to be target cells of the sex hormone-related dimorphism in the immune response peculiar to rheumatoid arthritis (RA) pathology. In the present study, the use of specific monoclonal antibodies revealed immunostaining for androgen and estrogen receptors in primary cultures of macrophages obtained from synovial tissues of patients affected by RA and controls without RA disease. Soluble and nuclear type I (high affinity, low capacity) and type II (lower affinity, greater capacity) sites of androgen or estrogen binding were detected in primary cultures of RA macrophages using radioligand binding assay. Higher levels of type I and type II estrogen receptor compared to those of androgen receptor were found, particularly in the soluble fraction; however, contrary to what was observed in whole synovial tissues, higher steroid receptor concentrations were found in the soluble than in the nuclear fraction of RA synovial macrophages. Binding affinities and receptor contents of cultured synovial macrophages were comparable to those previously reported in other well established sex hormone-responsive cells and tissues. Further, specific messenger ribonucleic acids for sex hormone receptors, encoding for a sequence of the DNA-binding domain of the receptor proteins were revealed by RT-PCR.

Adult

Sex steroids up-regulate E-cadherin expression in hormone-responsive LNCaP human prostate cancer cells.

There is convincing evidence that a reduced expression of the E-cadherin cell-cell adhesion molecule associates with low tumor grade and poor prognosis in prostate cancer patients. However, little is known on how E-cadherin levels are regulated in human prostate cancer cells. We have inspected the effect of both androgens and estrogen on the expression of E-cadherin in the hormone-responsive LNCaP prostate tumor cell line, which is endowed with both androgen and estrogen receptors. Using both Dot Blot analysis and immunocytochemistry we have observed that either steroid significantly increased E-cadherin levels in these cells; this effect was not reversed by the simultaneous addition of the relevant antagonist, hydroxyflutamide or ICI-182,780.

Androgen Antagonists

Distribution of GABA immunoreactivity in the retino-recipient layer of the viper optic tectum. A light and electron microscope quantitative study.

The distribution of GABA-immunoreactivity was investigated in the principal retino-recipient layer of the optic tectum in the snake Vipera aspis. This layer, the stratum griseum et fibrosum superficiale, contained an important proportion (approximately 50%) of small GABA-immunoreactive interneurons, characterized by a voluminous invaginated nucleus surrounded by a thin rim of cytoplasm poor in organelles and occasionally showing pleiomorphic synaptic vesicles, which could also be observed in some of the dendrites that contained synaptic vesicles. In the neuropile, the GABA-immunoreactive profiles containing synaptic vesicles could be subdivided into dendrites containing synaptic vesicles and axon terminals with pleiomorphic synaptic vesicles. The dendrites containing synaptic vesicles (23.4% of all profiles containing synaptic vesicles) were postsynaptic either to optic terminals (39.2%), GABA-immunoreactive axon terminals with pleiomorphic synaptic vesicles (48.2%) or to immunonegative (S1) boutons with round synaptic vesicles (12.6%). These dendrites were presynaptic to GABA-immunoreactive (18%) neurons or immunonegative (82%) neurons. The axon terminals with pleiomorphic synaptic vesicles, which represented 47.4% of all profiles, were predominantly (99%) GABA-immunoreactive and four types could be distinguished according to cytological criteria. These axon terminals made synaptic contacts for the most part (78%) with immunonegative profiles, and more rarely (22%) with immunoreactive neurons. These data are compared to those previously obtained in the homologous structure of other vertebrate species, birds and mammals in particular.

Animals

Ipsilateral visual projections in non-eutherian species: random variation in the central nervous system?

The published descriptions of ipsilateral visual pathways in non-eutherian species are reviewed. Such pathways exist in members of all vertebrate classes; since they exist in agnathans, it is suggested that the presence of ipsilateral visual projections is the ancestral condition. None of the published attempts to explain the considerable interspecific variation of these pathways can be generalised to all vertebrate species: in particular, this variation is not generally related to the degree of overlap of the visual fields, to a particular mode of life, nor to taxonomic position within a given vertebrate category and cannot consistently be explained by variation at the albino locus. It is suggested that this variation is the result of purely random variation of unidentified elements of the genetic material or of epigenetic mechanisms and hence that ipsilateral visual projections are functionally neutral. This conclusion is supported by some extremely fragmentary behavioral data indicating that the information they provide is redundant.

Animals

Anatomical evidence of a retino-thalamo-hippocampal pathway in the pigeon (Columba livia).

The aim of the present study was to demonstrate the existence of a retino-thalamo-hippocampal pathway in the pigeon and, more specifically, to identify its thalamic relay (n. superficialis parvocellularis: SPC). Fluorescence axonal tracing techniques were used combining the orthograde axonal transport of RITC after its intraocular injection and the retrograde axonal transport of various other fluorescent tracers (FB, FG, EB) injected into the hippocampal complex. Since the n.SPC has also been shown to project upon the visual Wulst, different fluorescent dyes were injected concomitantly into the latter and the hippocampus and the distribution of retrograde somatic labeling within the thalamus was compared. The injections into both telencephalic target structures resulted in a differential pattern of labeling bilaterally within the thalamic dorsolateral anterior (DLA) complex and the n.SPC. Moreover, the distributions of both cell types overlapped within the n.SPC, although no double-labeling of individual cell bodies indicative of collateralized axonal projections was observed. Direct retinal projections upon the n.SPC were also demonstrated and terminated within the same regions of the nucleus containing relay cells projecting to both the hippocampus and the Wulst. The organization of the retino-thalamo-hippocampal pathway in birds is similar to that which has been described in other vertebrate groups. A common feature is the presence of divergent projections from a dorsal thalamic retino-recipient relay nucleus upon both neopallial visual cortical areas and the hippocampus (archipallium).

Animals

The distribution of GABA-immunoreactive neurons in the brain of the silver eel (Anguilla anguilla L.).

The distribution of GABA-immunoreactivity was studied in the brain of the silver eel (Anguilla anguilla) by means of antibodies directed against GABA. Immunoreactive neuronal somata were distributed throughout the brain. Positive perikarya were detected in the internal cellular layer of the olfactory bulb, and in all divisions of the telencephalon, the highest density being observed along the midline. Numerous GABA-reactive cell bodies were found in the diencephalon, particularly in the preoptic and tuberal regions of the hypothalamus, and the dorsolateral, dorsomedial and ventromedial thalamic nuclei. In the optic tectum, the majority of GABA-positive cell bodies were located in the periventricular layer. A number of immunolabeled cell bodies were observed in different tegmental structures, notably the torus semicircularis. In the cerebellum, the Purkinje cells were either very intensely or very weakly immunoreactive. In the rhombencephalon, reactive cell bodies were observed in the eminentia granularis, the valvula cerebellaris, the octavolateral nucleus, the lobus vagus and in the vagal and glossopharyngeal motor nuclei. Intensely immunoreactive axons and terminals were observed in the external granular layer and internal cellular layer of the olfactory bulb. In the telencephalon, the highest density of reactive fibres and boutons was found in the fields of the medial wall. Many immunolabeled fibres were seen in the medial and lateral forebrain bundles. In the diencephalon, intense labelling of fibres and terminals were observed in the nuclei situated close to the midline. In the optic tectum the highest density of reactive fibres was seen in the sfgs, the layer to which the retina projects massively. Finally, in the rhombencephalon the strongest labelling of neurites was observed in the nuclei of the raphé, the nucleus octavocellularis magnocellularis and the nuclei of the IXth and Xth cranial nerves. The GABAergic system of the eel, which is well developed, appears to be generally comparable to that described in tetrapod vertebrates.

Anguilla

Retrograde transneuronal transport of the fluorescent dye rhodamine beta-isothiocyanate from the primary and centrifugal visual systems in the pigeon.

The transneuronal labeling properties of the fluorescent dye Rhodamine beta-isothiocyanate (RITC) were investigated in the pigeon following the intraocular injection of the tracer either alone or in combination with kainic acid (RITC/KA). The RITC was transported bidirectionally from the eye producing both orthograde axonal and terminal labeling of the primary visual system (PVS) and retrograde labeling of the centrifugal visual system (CVS) comprising the n. isthmo-opticus (NIO) and associated ectopic neurons (EN) as well as the tractus isthmo-opticus (TIO). In addition, RITC-positive cell bodies were identified in layers 9/10 of the optic tectum, n. isthmi parvocellularis (Ipc) and the hyperstriatum accessorium (HA) subdivision of the telencephalic visual Wulst. In the RITC/KA experiments, the orthograde transport of the dye within the PVS was selectively suppressed and this coincided with the absence of somatic labeling in n. Ipc and HA. The results are explained in terms of transneuronal transport of RITC, involving the terminal uptake and subsequent retrograde axonal transport of the tracer within second-order neurons in n. Ipc and HA which project to the PVS and in tectal layers 9/10 projecting to the CVS. Moreover, the transneuronal transport of RITC, as demonstrated using the present experimental conditions, appears to be very specific and only labeled distant cell populations known to project to either the PVS or CVS.

Animals

Evidence of GABA-immunopositive neurons in the dorsal part of the lateral geniculate nucleus of reptiles: morphological correlates with interneurons.

The distribution and staining pattern of gamma-aminobutyric acid immunoreactivity have been examined by both light and electron microscopy in the dorsal part of the lateral geniculate nucleus of three reptilian species: the turtle Chinemys reevesi, the lizard Ophisaurus apodus and the snake Vipera aspis. After perfusion of the animals with 1% paraformaldehyde and 1% glutaraldehyde and polyethyleneglycol embedding of the brains, the analysis of sections processed immunocytochemically with an anti-GABA antiserum has revealed a moderate-to-dense labeling of the neurons of the dorsal part of the lateral geniculate complex in these species. Labeled cell bodies are small-sized, either rounded or fusiform and the GABA-positive dendrites emerging from them are not preferentially oriented in any particular direction. Quantitative studies in Vipera indicate that GABA-positive neurons make up about 14% of the population of neurons of the dorsal part of the lateral geniculate nucleus. Electron microscopy of specimens treated by either pre- or post-embedding techniques has confirmed that these cells corresponded to neurons. No glial cells were ever observed to be immunopositive. These GABA-positive neurons, characterized by the presence of pleiomorphic synaptic vesicles localized either in their perikaryon or more often in presynaptic dendrites, established symmetrical synaptic contacts. In this case, the latter were involved both pre- and postsynaptically in serial and, more rarely, in triadic arrangements, a synaptic organization specific to interneurons. The involvement of such GABA-positive neurons in local circuits is discussed.

Animals

Divergence and collateral axon branching in subsystems of visual cortical projections from the cat lateral posterior nucleus.

The projections from the lateral (LPl), intermediate (LPi) and medial (LPm) subdivisions of the cat lateral posterior nucleus (n. LP) to visual areas 17, the posteomedial (PMLS) and posterolateral (PLLS) lateral suprasylvian and anterior ectosylvian (AEV) were studied using the retrograde labeling technique following concomitant injections of fluorescent dyes (Fast blue, Nuclear yellow, Evans blue and Rhodamine beta-isothiocyanate) into the different cortical loci. The results showed a medial-lateral topographical reversal of the visual n. LP-cortical connections: The ventral portion of LPl projects to area 17 whereas more dorsolateral regions of LPl and lateral LPi provide input to PMLS. Cells in medial LPi project mainly to the PLLS cortex and AEV receives afferents from the LPm. Areas of overlap were identified within the ventral LPl which projects to both area 17 and PMLS and within the LPi/LPm border region at the origin of connections to both PLLS and AEV. Furthermore, some single neurons within the areas of overlap were found to be double-labeled indicating divergent projections to their respective cortical targets via collateral axon branching. The data show that divergence and axonal branching are common features of the different n.LP-visual cortical subsystems and support the notion of the existence of families of thalamo-cortical systems which are distinct in their connection patterns and underlying functional properties.

Animals