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N Corvaja

Publications and source records attributed to N Corvaja.

At least 37 records · Page 2Linked to original sources

Ultrastructure and synaptic targets of the raphe-nigral projection in the rat.

A major input to the substantia nigra is from the 5-hydroxytryptamine-containing neurons in the dorsal raphe nucleus. In order to examine the morphology and distribution of this projection, rats were given injections of the anterograde tracers, Phaseolus vulgaris-leucoagglutinin or biocytin, in the dorsal raphe nucleus and the substantia nigra was examined at both the light and electron microscopic levels. In addition, sections of the substantia nigra were immunostained for 5-hydroxytryptamine and examined in both the light and electron microscopes. Since dopaminergic neurons of the substantia nigra are known to be responsive to stimulation of the raphe and to applied 5-hydroxytryptamine, sections that contained anterogradely labelled terminals were further processed to reveal tyrosine hydroxylase immunoreactivity to determine whether the raphe input makes direct synaptic contact with dopaminergic neurons. Light microscopic analysis revealed that all divisions of the substantia nigra received input from the dorsal raphe which, in agreement with previous observations, showed a topographical organization. In that formed asymmetrical synaptic contact with dendritic shafts and spines. The synaptic boutons were often associated with subjunctional dense bodies. Terminals that displayed immunoreactivity for 5-hydroxytryptamine had a similar morphology, synaptic specialisations and postsynaptic targets to the anterogradely labelled terminals. In those sections that were stained for both anterogradely labelled terminals and tyrosine hydroxylase, the raphe-nigral terminals were seen to form asymmetrical synaptic contact with the dendrites of the dopaminergic neurons. It is concluded that dendrites of dopaminergic neurons in the substantia nigra pars reticulata represent at least one of the synaptic targets of the raphe-nigral projection and that these contacts provide an anatomical substrate for the effects of the dorsal raphe, and presumably 5-hydroxytryptamine, on dopaminergic systems in the substantia nigra.

Animals↗

Immunocytological localization by monoclonal antibodies of alpha-latrotoxin in the venom gland of the spider Latrodectus tredecimguttatus.

Monoclonal antibodies, raised against alpha-latrotoxin, have been used on serial sections of the venom glands with localization by indirect immunofluorescence. Colloidal gold immunovisualization and electron microscopy were used in order to localize the structure of the cells that synthesize and store the toxin. The antibodies were able to recognize the native toxin revealing its presence mainly in the 'replacement cell' layer, a layer of cells close to the muscular sheath of the gland. These cells apparently replace the disintegrated epithelial cells at the end of the holocrine cycle. Gold grains were predominantly observed around the free ribosomes and the secretion droplets of the 'replacement cells'.

Animals↗

Intracellular localization of nerve growth factor in PC12 cells. An immuno-gold electron microscopic approach.

Nerve growth factor (NGF) localization in the clonal cell line PC12 has been assessed with the immunocytochemical colloidal gold staining (IGS) method. Three distinct fixation and staining procedures have been adopted to avoid possible artifactual localizations and to better assess NGF intracellular targets. Colloidal gold particles are observed in the cytoplasm and also within the nuclear compartment, close to the nuclear envelope. Gold particles are also detectable, in aggregates or patches, in proximity of or in association with, cytoskeletal elements tentatively identified with microfilaments (MF) present in the growth cone and microtubules (MT) running in parallel within the neurites or present in the perikaryon. Neurofilaments (NF), on the contrary, were devoid of any NGF-antibody cross-reacting material detectable with colloidal gold staining. It is postulated that the association of NGF or NGF-receptor complexes with cytoskeletal elements is instrumental for the mechanism of action of NGF.

Animals↗

Nuclear localization of a lactic dehydrogenase with single-stranded DNA-binding properties.

In the preceding article [1] we identified the 34 kD single-stranded DNA-binding (ssb) protein, whose synthesis is inhibited in PC12 cells concomitantly with nerve growth factor (NGF)-induced mitotic arrest, with the enzyme lactic dehydrogenase (LDH-ssb protein). Localization studies performed with antibodies raised against the LDH-ssb protein demonstrate the presence of a pool of this protein in the nucleus of several cell types. The nuclear association of this protein is sensitive to DNase treatment of the cells and quantitative electron microscopy confirms that the LDH-ssb protein is located close to chromatin structures. These results point to a possible involvement of the LDH-ssb protein in some nuclear function(s).

Animals↗

Cerebellar output regulation by the climbing and mossy fibers with and without the inferior olive.

The activity of the olivocerebellar complex and the structures related in series with it have been studied using the complementary action of harmaline and 3-acetylpyridine to isolate the two principal inputs to the cerebellar Purkinje cells. The activities of the various nuclei as well as the entire brain have been simultaneously monitored using the [14C]2-deoxy-glucose method under the various combined effects of the pharmacological agents. (1) Tremogenic doses of harmaline increased the frequency of discharge in selected parts of the olivocerebellar system, increasing climbing fiber input and reducing Purkinje cell simple spike discharges in corresponding parts of the cerebellar cortex. The metabolic activity increased in the inferior olive and in the red nucleus. The results are interpreted as a net reduction of Purkinje cell inhibition on their target neurons, leading to a facilitatory cerebellar output. (2) Systemic injection of neurotoxic doses of 3-acetylpyridine selectively produced total degeneration of the neurons in the inferior olive, resulting in the suppression of complex spikes and a net increase in simple spike output from the Purkinje cells. The metabolic consequences were a reduction or absence in the inferior olive, decrease in the red nucleus, and increases in the Purkinje cell target neuron regions, including the intracerebellar and vestibular nuclei. The study of long survival times following the neurotoxic treatment revealed a transient metabolic marking of the inferior olive during the active glial processes accompanying the degeneration. In other parts the radioautographic changes caused by the destruction of the inferior olive persisted for about 1 month after the administration of the drug. (3) Tremogenic doses of harmaline were given to rats at different times following treatment with 3-acetylpyridine. It was demonstrated that: (a) intoxication of the inferior olive started within the second hour after 3-acetylpyridine administration, corresponding to the time at which the metabolic response to harmaline was also abolished; and (b) the increased metabolic activity produced by harmaline in the olivocerebellar complex was a consequence of an increased activity of the neurons of the inferior olive rather than a direct pharmacological effect of the drug. (4) Partial lesions of the inferior olive led to increased metabolic activity of those parts of the intracerebellar nuclei topographically related to the destroyed parts of the inferior olive. (5) In 3-acetylpyridine-treated animals, local ablation as well as local inactivation of the cerebellar cortex produced localized suppression of the intense labeling in the intracerebellar nuclei obtained in these animals. Since these regions receive synapses which are normally inhibitory, suppression of labeling clearly supports the hypothesis that regional marking may very well be produced by the activity of the presynaptic terminals themselves...

Animals↗

Spinal projections from the mesencephalon in the toad.

Mesencephalic cell groups projecting to the spinal cord have been identified by means of the retrograde axonal transport of the enzyme horseradish peroxidase (HRP). The injections were made either in the cervical or lumbar enlargements of the toad spinal cord. Following injections in the cervical cord, labeled cells are located in the isthmus region, in the ipsilateral laminated nucleus posteroventralis tegmenti mesencephali (Potter). At more rostral levels the labeled cells are in the nucleus of the fasciculus longitudinalis medialis, in the nucleus interstitialis of the fasciculus longitudinalis medialis, in the contralateral red nucleus, in lamina six of the contralateral optic tectum, bilaterally in the nucleus of the posterior commissure and in the mesencephalic nucleus in the Vth nerve. Injections in the lumbar cord label neurons of the nucleus posteroventralis tegmenti mesencephali (Potter) and nucleus interstitialis of the fasciculus longitudinalis medialis. Nuclei that had not been previously identified in anurans but which were labeled after HRP spinal injections (i.e., the nucleus interstitialis of the fasciculus longitudinalis medialis, the nucleus of the posterior commissure and the red nucleus) have been delimited in normal material in Nissl-stained transverse sections. The spinal pathways from the mesencephalon can be classified into four projections: reticulospinal, rubrospinal, tectospinal and trigeminospinal. A comparison of these descending fiber systems with homologous pathways in other vertebrate species has been made.

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Spinal projections from the rhombencephalon in the toad.

Rhombencephalic cell groups projecting to the spinal cord are demonstrated following single pressure injections and/or iontophoretic ejections of HRP solution in either cervical or lumbar enlargements of the toad spinal cord. A group uptake and transport of HRP were obtained with both application techniques, when sufficiently long survival times (8-11 days) were used. Following injections in the cervical cord labeled cells are located mostly in the ventral nucleus of the VIIIth nerve and in the medial zone of the rhombencephalic reticular formation, i.e. the nucleus reticularis inferior, medius and superior. Following injections in the lumbar enlargement the majority of labeled cells are situated in the caudalmost portion of the ventral nucleus of the VIIIth nerve and in the nucleus reticularis inferior. These observations indicate that in the toad the main supraspinal descending pathways from the rhombencephalon originate in the ventral nucleus of the VIIIth nerve and the medial zone of the reticular formation, and that both these pathways are somatotopically organized.

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[Somatotopic organization of the vestibulospinal tract in the toad].

The origin of the vestibulospinal projection in the toad has been investigated by using the method of the retrograde axonal transport of HRP injected at various levels of the spinal cord. The vestibulospinal projection, in this species, was found to be somatotopically organized, since neurons projecting to the cervical segments of the spinal cord were located within the rostromedial part of the ventral vestibular nucleus and those neurons projecting to the lumbosacral segments of the spinal cord were located within the caudolateral part of that nucleus. This pattern of organization of the vestibulospinal projection in amphibia is similar to that described in mammals and birds.

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Ultrastructure of supraspinal dorsal root projections in the toad. II. The cerebellar granular layer.

Following section of the left dorsal roots, degenerating fibres and boutons were observed in the granular layer of the ipsilateral cerebellum. The degenerating terminals were identified as large en passant varicosities of mossy fibres contacting the dendrites of presumptive granule cells. They contained round synaptic vesicles and neurofilaments and established Gray type I contacts. The terminals initially underwent filamentous degeneration with neurofilamentous hypertrophy, swollen mitochondria and loss of synaptic vesicles. At later survival times (6--30 days) they acquired an electron-dense appearance due to an increase and clumping of the filamentous component. After injection of horseradish peroxidase into the left cerebellum, all ipsilateral spinal ganglia showed a few (2--3%) labelled cells, indicating that a primary afferent contribution to this pathway originated from each segment of the spinal cord.

Afferent Pathways↗

Ultrastructure of supraspinal dorsal root projections in the toads. I. The obex region.

The ultrastructure of the dorsal column nucleus (DCN) has been investigated at the level of the obex region in normal and experimental toads. Large 'isolated' neurons (greater than 20 micrometer) and clusters of small neurons (less than 20 micrometer) have been identified in this region. Synaptic profiles have been classified into three types: large 'en passant' LR boutons, containing round synaptic vesicles and neurofilaments, small R boutons with round vesicles and F boutons with pleomorphic vesicles. The axon terminals exhibited synaptic contacts with cell somata, with dendrites of varying calibers and with other axons. The terminals involved in the axo-axonic contact were the F boutons which were presynaptic to the LR boutons, thus representing the morphological basis for presynaptic inhibition. Transection of the second dorsal root was performed in order to identify the terminals of the primary afferents to the DCN, after different survival periods (16 h--50 days). Only the LR boutons underwent degeneration, thus representing the central endings of the primary dorsal root afferents. The functional significance of these findings was discussed.

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Identification of different size motoneurons labeled by the retrograde axonal transport of horseradish peroxidase.

Three main groups of motoneurons of different size have been labeled in adult cats by using the method for retrograde axonal transport following injection of horseradish peroxidase in the medial gastrocnemius and soleus muscles. In particular small, medium-size and large neurons which probably correspond respectively to gamma, small alpha and large alpha motoneurons innervating the calf muscles, have been identified and the corresponding area measured.

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