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E Mugnaini

Publications and source records attributed to E Mugnaini.

At least 91 records · Page 5Linked to original sources
PubMed1985

GABA neurons in the superficial layers of the rat dorsal cochlear nucleus: light and electron microscopic immunocytochemistry.

This article is an application of light and electron microscopic immunocytochemistry to the study of the neuronal circuit of the superficial layers in the rat dorsal cochlear nucleus (DCN). An antiserum against the intrinsic marker glutamate decarboxylase (GAD) is used to identify and map axon terminals and neurons that use gamma aminobutyric acid (GABA) as a neurotransmitter. It is demonstrated that layers 1 and 2 of the DCN contain a very high density of GABAergic boutons, matched only by the granule cell domains of the ventral cochlear nucleus, especially the superficial granule cell domain. These two layers also contain much higher concentrations of GABAergic cell bodies than all other magnocellular regions of the cochlear nuclear complex. Cartwheel and stellate neurons, and probably also Golgi cells, previously characterized in Golgi and electron microscopic investigations, appear immunostained and, therefore, are presumably inhibitory. The synaptic relations between parallel fibers, the axons of granule cells, and cartwheel and stellate neurons are confirmed. The present study also supports the conclusion that stellate cells are coupled to one another by gap junctions. Also scattered in layer 1 are large, GABAergic neurons that occur with irregular frequency and presumably represent displaced Purkinje cells, previously identified with a Purkinje-cell-specific marker. Granule neurons and pyramidal neurons remain unstained, even after topical injection of colchicine, which enhances immunostaining of the other glutamate-decarboxylase-positive cells, and therefore must use transmitters different from GABA. The possible analogies between the spiny cartwheel and the aspiny stellate cells of the DCN and the cerebellar Purkinje and stellate/basket cells are discussed in the light of data from Golgi, electron microscopy, and transmitter imunocytochemistry.

Animals↗
PubMed1985

Cell-cell junctional interactions and characteristic plasma membrane features of cultured rat glial cells.

Mixed cultures of astrocytes and oligodendrocytes derived from cerebral hemispheres of 18-19 day old rat fetuses were studied with the freeze-fracture technique. The plasma membranes of cultured astrocytes and oligodendrocytes differ substantially in their intramembrane particle profiles, and they can be positively identified consistently. Orthogonal small particle assemblies and numerous isolated globular particles characterize astrocytic plasma membranes, whereas the plasma membranes of oligodendrocytes show numerous elongated particles and fewer large and small globular particles similar to those seen in situ. Using these distinct differential features, we can identify partners of glial cell junctions. We can identify numerous interastrocytic gap junctions, as well as heterologous astrocyte-to-oligodendrocyte gap junctions. The plasma membranes of adjacent oligodendrocytes form numerous tight junctions consisting of linear P face strands and/or rows of particles interrupted by short segments of grooves, the complementary features on the E face. "Reflexive" type tight junctions seen in situ are also observed. In addition to intercellular junctions, glial cells develop special plasma membrane structural domains. Astrocytic plasma membranes often contain groups of plasmalemmal vesicles (caveolae), a distinctive feature of astrocytes in situ. Oligodendrocytes form flattened velate processes with cytoplasm restricted to finger-like channels resembling myelin lamellae in situ. Cultured astrocytes and oligodendrocytes develop the entire range of plasma membrane structural specializations seen in situ in the absence of the normal brain tissue framework. Thus, primary glial cell cultures allow experimental study of many glial cell properties, including their plasma membrane specializations.

Animals↗
PubMed1984

Cartwheel neurons of the dorsal cochlear nucleus: a Golgi-electron microscopic study in rat.

Cartwheel neurons in rat dorsal cochlear nucleus (DCN) were studied by Golgi impregnation-electron microscopy. Usually situated in layers 1-2, cartwheel neurons (10-14 micrometers in mean cell body diameter) have dendritic trees predominantly in layer 1. The dendrites branch at wide angles. Most primary dendrites are short, nontapering, and bear only a few sessile spines. Secondary and tertiary dendrites are short, curved, and spine-laden. The perikaryon forms symmetric synapses with at least two kinds of boutons containing pleomorphic vesicles. The euchromatic nucleus is indented and has an eccentric nucleolus. The cytoplasm shows several small Nissl bodies, a conspicuous Golgi apparatus, and numerous subsurface and cytoplasmic cisterns of endoplasmic reticulum with a narrow lumen, joined by mitochondria in single or multiple assemblies. In primary dendrites mitochondria are situated peripherally, while in distal branches they become ubiquitous and relatively more numerous. Dendritic shafts usually form symmetric synapses with boutons that contain pleomorphic vesicles. The majority of the dendritic spines are provided with a vesiculo-saccular spine apparatus. All dendritic spines have asymmetric synapses. Most of these are formed with varicosities of thin, unmyelinated fibers (presumably axons of granule cells) running parallel to the long axis of the DCN or radially. These varicosities contain round, clear synaptic vesicles. On the initial axon segment few symmetric synapses are present. The axon acquires a thin myelin sheath after a short trajectory. Cartwheel neurons outnumber all other neurons in layers 1-2 (with the exception of granule cells), and presumably correspond to type C cells with thinly myelinated axons described by Lorente de Nó. The axons of these neurons provide a dense plexus in the superficial layers without leaving the DCN. The possible functional role of cartwheel neurons is discussed.

Animals↗
PubMed1984

Immunocytochemical localization of GABA neurons and dopamine neurons in the rat main and accessory olfactory bulbs.

Immunocytochemical localization of GABA neurons and dopamine neurons in the rat olfactory bulb was obtained with sheep antiserum to glutamate decarboxylase (GAD) and rabbit antiserum to tyrosine hydroxylase (TH). GAD-positive neurons include periglomerular cells, granule cells, superficial and deep short axon cells. TH-positive neurons represent periglomerular cells. Two-color immunocytochemistry shows that GABA and dopamine periglomerular cells are separate populations. The accessory olfactory bulb has rare dopamine cells and few superficial short axon cells. Radial gradients of GAD-immunostaining are evident in the main but not in the accessory olfactory bulb.

Animals↗
PubMed1984

Immunocytochemical studies of GABAergic neurons in rat basal ganglia and their relations to other neuronal systems.

GABAergic neurons were localized in the rat basal ganglia by glutamate decarboxylase (GAD) immunohistochemistry. In the striatum (caudato-putamen, accumbens nucleus) a medium density of GAD-positive terminals was observed; a small number of medium-to-large size neurons and the vast majority of medium-size neurons were GAD immunoreactive. In addition, opioid peptide-like immunoreactivity was colocalized in a subclass of GAD-positive medium-size striatal neurons. The pallido-nigral system (GP, VP, EP, SNR) displayed a high density of GAD-positive axon terminals which synapsed upon dendrites and nerve cell bodies. The majority of pallido-nigral neurons also were GAD-immunoreactive. In contrast, the substantia nigra pars compacta and the subthalamic nucleus contained only few GAD-immunoreactive neurons.

Animals↗
PubMed1984

Stellate neurons in rat dorsal cochlear nucleus studies with combined Golgi impregnation and electron microscopy: synaptic connections and mutual coupling by gap junctions.

Stellate neurons in the outer two layers of the rat dorsal cochlear nucleus (DCN) were studied by the Golgi-EM method. Stellate cell bodies are usually spherical or ovoidal and range from 9 microns to 14 microns in mean diameter. The smallest cells are situated underneath the ependymal layer and the largest cells in layer 2. Primary dendrites are short, thin and smooth and arise abruptly from the perikaryon, without a tapering main stem. Meandering secondary and tertiary dendrites extend in all directions, carry few pleomorphic spines lacking a spine apparatus and often show artifactual beading. The axons are impregnated only for a short distance (10-45 microns). The nucleus is indented, the nucleolus varies in position, and the chromatin, evenly dispersed in the centre, forms small clumps along the nuclear envelope. The cytoplasm is rich in free polyribosomes and contains scattered cisterns of granular endoplasmic reticulum. Varicosities of thin fibres, containing round synaptic vesicles, form asymmetric synapses on perikarya, dendritic shafts and spines of stellate cells. Such fibres run parallel to the long axis of the DCN or are oriented radially and are interpreted as axons of cochlear granule cells. Two kinds of bouton containing pleomorphic vesicles, one kind electron lucent and the other electron dense, form symmetric synapses on perikarya and dendritic shafts of stellate cells. The lucent boutons occur more frequently than the dense boutons, especially on the distal dendritic branches. The boutons with pleomorphic vesicles presumably represent terminals of local circuit neurons, probably the stellate and cartwheel cells. In addition, stellate cells show numerous dendro-somatic and dendro-dendritic appositions characterized by gap junctions and puncta adhaerentia. Most of the dendrites involved in these appositions resemble stellate cell dendrites and it is concluded that DCN stellate cells are coupled electrotonically with one another. The axons of stellate cells acquire a thin myelin sheath. Since the Golgi impregnation did not stain axons of stellate cells past this point, we were unable to demonstrate the synaptic targets of stellate cells.

Animals↗
PubMed1984

Pyramidal neurones of the dorsal cochlear nucleus: a Golgi and computer reconstruction study in cat.

The main projection neurones of the dorsal cochlear nucleus, termed pyramidal, bipolar or fusiform cells, have an apical dendritic arbor approaching the ependymal surface of the nucleus and a basal arbor oppositely directed. In Golgi-Del Rio-Hortega material these neurones were studied, with the light microscope, in nonconventional planes of sectioning oriented across or parallel to the main axis of the elongated nucleus. The pyramidal neurones were seen to be flattened across this axis. The size, shape and orientation of 21 cells from six blocks were studied in detail with computer-aided graphic reconstructions including stereo views. Camera lucida drawings of each cell (usually from several sections) were digitized to obtain x and y coordinates while z coordinates (depths in the tissue) were read from the fine focus knob during microscopy and typed interactively during digitization. The z values were corrected for the effects of refractive index differences in the optical system. Since it was the aim of this study to focus on some fundamental principles of structure and arrangement of pyramidal cells in the dorsal cochlear nucleus rather than on topographic variations, only the middle, regularly built part of the nucleus was examined. Towards the ends of the nucleus the architecture is less regular and will require separate analysis. Measurements of arbor and total cell height and of dendritic length are given. The height of the apical and basal arbor in individual cells showed considerable reciprocity. The total dendritic length was up to 8300 micron (average 6536 micron). The basal arbors always proved to be conspicuously flattened; roughly, the width varied between about 300 and 700 micron (average 489 micron) and the thickness between 65 and 105 micron (average 80 micron). The apical arbors were also often flattened but much less and with a greater variability than the basal arbors (average width 319 micron, thickness 115 micron). The two arbors of individual cells were practically coplanar, the arbor planes showing only moderate angularity (bend) and/or torsion relative to each other (angularity maximum 10 degrees, average 5 degrees; torsion maximum 18 degrees, average 6 degrees). The mutual orientation of cells from the same block was examined. The planes through the basal arbors proved to be very parallel, the differences in orientation angles being between 10 and 0 degrees with rare exceptions. Clearly flattened, apical arbors showed a somewhat greater spread.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1984

Dorsal nucleus of the lateral lemniscus: a nucleus of GABAergic projection neurons.

Immunocytochemical staining of the dorsal nucleus of the lateral lemniscus with a well characterized antiserum to glutamate decarboxylase reveals that all, or nearly all, cells in this nucleus show immunoreactivity without the use of agents to block axonal transport. Most somata and dendrites are also contacted by immunoreactive axonal endings. It has previously been established that this nucleus is richly innervated by ascending lemniscal fibers and contains different types of neurons that project to the inferior colliculus. One may conclude that this precollicular nucleus is a major GABAergic feedforward inhibitory center in the acoustic pathway.

Animals↗
PubMed1984

Immunocytochemical demonstration of the GABA-ergic neurons in rat globus pallidus and nucleus entopeduncularis and their GABA-ergic innervation.

GAD (E.C. 4.1.1.15), the endogenous biosynthetic marker enzyme for GABA, was localized in rat globus pallidus and nucleus entopeduncularis (pallidum) by peroxidase-antiperoxidase immunocytochemistry. In the two nuclei we observed numerous GAD-like immunoreactive boutons containing large pleomorphic vesicles. These boutons formed axodendritic and axosomatic synaptic contacts mostly of the symmetric type. Following colchicine injection near the border of globus pallidus or nucleus entopeduncularis, the vast majority of both larger and smaller neurons exhibited GAD immunoreactivity and were contacted by numerous GAD-immunoreactive boutons. These data demonstrate that pallidal neurons are of predominately GABA-ergic nature and receive an extensive GABA-ergic input. Thus, the globus pallidus and the nucleus entopeduncularis major output stations of the basal ganglia may represent sites of extensive GABA-mediated inhibition of GABA-ergic inhibitory neurons.

Animals↗
PubMed1984

Immunocytochemical identification of GABAergic neurons in the main olfactory bulb of the rat.

With the aid of a sheep antiserum against rat brain glutamate decarboxylase (GAD), the endogenous marker for GABAergic neurons, we have labeled immunocytochemically various types of nerve cells in the main olfactory bulb of rats, with and without topic injections of colchicine. The peroxidase-antiperoxidase procedure was applied to floating Vibratome and frozen sections. A large part of the periglomerular cell population and practically all granule cells in the deep layers contain GAD-like immunoreactivity in untreated rats, while tufted and mitral cells (the projection neurons) are unstained. This observation confirms a previous study with a rabbit antiserum against mouse brain GAD, which suggested that GABAergic neurons with presynaptic dendrites contain high somatal concentrations of GAD. We show, however, that immunostaining of granule cell bodies decreases progressively from the internal plexiform layer to the deep portion of the granule cell layer. Many cell processes in the glomeruli are densely stained. They presumably represent synaptic gemmules of the numerous GAD-positive periglomerular cells, which thus could provide initial, inhibitory modulation of the afferent input. In the external plexiform layer immunostaining of the neuropil is substantially denser in the superficial half than in the deep half. This may reflect a corresponding gradient of inhibition related to unequal frequency of occurrence of synaptic gemmules of granule cell dendrites. Alternatively such a graded immunostaining of cell processes could be related to the corresponding gradient in the density of immunostaining of granule cell bodies in the deep layers, in accordance with recent data indicating that superficial and deep granule cells project their ascending dendrites respectively to superficial and deep portions of the external plexiform layer. Furthermore, we have demonstrated the presence of additional classes of GAD-positive neurons, microneurons in the external plexiform layer, small neurons in the periglomerular region, the external plexiform layer, the mitral cell layer, the internal plexiform layer, and medium-size neurons in the granule layer and the white matter. The small- and medium-size GAD-positive neurons appear weakly immunoreactive in untreated rats, but become densely stained after topic colchicine injection. Such cells presumably lack presynaptic dendrites and may correspond to different types of short axon cells demonstrated by the Golgi method.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1984

Cell-cell interactions of isolated and cultured oligodendrocytes: formation of linear occluding junctions and expression of peculiar intramembrane particles.

Oligodendrocytes were isolated from lamb brain. Freshly isolated cells and cultured cells, either 1- to 4-day-old unattached or 1- to 5-week-old attached, were examined by thin section and freeze-fracture electron microscopy. Freeze-fracture of freshly isolated oligodendrocytes showed globular and elongated intramembrane particles similar to those previously described in oligodendrocytes in situ. Enrichment of these particles was seen at sites of inter-oligodendrocyte contact. Numerous gap junctions and scattered linear tight junctional arrays were apparent. Gap junctions were connected to blebs of astrocytic plasma membrane sheared off during isolation, whereas tight junctions were facing extracellular space or blebs of oligodendrocytic plasma membrane. Thin sections of cultured, unattached oligodendrocytes showed rounded cell bodies touching one another at points without forming specialized cell junctions. Cells plated on polylysine-coated aclar dishes attached, emanated numerous, pleomorphic processes, and expressed galactocerebroside and myelin basic protein, characteristic markers for oligodendrocytes. Thin sections showed typical oligodendrocyte ultrastructure but also intermediate filaments not present in unattached cultures. Freeze-fracture showed intramembrane particles similar to but more numerous, and with a different fracture face repartition, than those seen in oligodendrocytes, freshly isolated or in situ. Gap junctions were small and rare. Apposed oligodendrocyte plasma membrane formed linear tight junctions which became more numerous with time in culture. Thus, cultured oligodendrocytes isolated from ovine brains develop and maintain features characteristic of mature oligodendrocytes in situ and can be used to explore formation and maintenance of tight junctions and possibly other classes of cell-cell interactions important in the process of myelination.

Animals↗
PubMed1984

Histochemical localization of acetylcholinesterase in the cochlear and superior olivary nuclei. A reappraisal with emphasis on the cochlear granule cell system.

The present account reconsiders the acetylcholinesterase (AChE) staining pattern of the cochlear nuclei with special emphasis on positively stained afferents from the olivocochlear neurons to the cochlear granule cells system. The main part of the study is based on AChE- and silver-stained sections of normal and brainstem-operated cats. AChE-stained sections of normal mouse, cat and chinchilla are used for comparative purposes. The rat superior olive contains three types of AChE-positive neurons probably contributing to the olivocochlear bundle; densely stained large neurons of the periolivary region, densely stained small neurons at the margin of the lateral superior olive (LSO), and weakly stained small neurons within LSO. Largely uncrossed fibers, probably collaterals of the olivocochlear bundle, enter the cochlear nuclei via three routes, defined here as the strial, subpeduncular and ventral routes. Collectively they form a terminal fiber plexus in certain portions of the granule cell domain, but with some fibers branching in the extragranular regions of the nuclear complex as well. The individual fibers end in a fashion resembling cerebellar mossy fibers. The cochlear nuclei in addition contain conspicuous dense patches of precipitate, which, like the AChE-positive fibers, appear after short incubation and are mainly restricted to the granule cell domain. In contrast to the fibers, however, they are resistant to central deafferentation and therefore may represent intrinsic structures of the granule cell system. Moreover, there is a diffuse neuropil precipitate which grows in distribution and density with incubation time. It is present both in granular and extragranular areas of the complex and is partly dependent upon the integrity of the weakly stained trapezoid body. In spite of considerable interspecies variations with regard to the described AChE-positive elements, these are present also in mouse, cat, and chinchilla. The significance of the findings and of the interspecies differences are discussed. A simplified terminology of the superior olivary complex is proposed.

Acetylcholinesterase↗
PubMed1983

The length of cerebellar parallel fibers in chicken and rhesus monkey.

The cerebellar parallel fibers, which course through the molecular layer parallel to the long axes of the cortical folds known as folia, originate from ascending granule cell axons and relay the mossy fiber input to dendrites of Purkinje cells. Purkinje cell axons in the cerebellar white matter collect into sheets or zones oriented at right angles to the folia. Each of these zones, which are approximately 0.5-1 mm wide, innervates a different portion of the deep cerebellar and the vestibular nuclei. An experimental light microscopic study was carried out to determine the maximal length of parallel fibers in long folia of avian and primate cerebellar cortex. With a fine surgical knife, vermal folia were cut perpendicular to their long axes in four adult White Leghorn hens and in three adult rhesus monkeys deeply anesthetized with sodium pentobarbital. The animals were Killed 3-5 days after the operation. Sections of the transected folia were stained with the Fink-Heimer or the DeOlmos-Ingram methods, which revealed the anterogradely degenerated parallel fibers as darkly stained dots. In both species, the pattern of parallel fiber degeneration in the molecular layer had a trapezoidal configuration with the shorter base bordering the Purkinje cell layer and the longer base bordering the pia mater. In both species, the length of parallel fibers averaged approximately 6 mm, although the range was 4-8 mm in chickens and 4.8-6.6 mm in monkeys.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1983

Myelin sheath thickness in the CNS is regulated near the axon.

Two myelin sheaths sharing a common outer tongue process are illustrated in a thin section electron micrograph from mouse optic nerve. Although they derive from the same outer tongue process, and therefore from the same oligodendrocyte, the two sheaths are of different thicknesses (6 and 9 turns). The example demonstrates that myelin sheath thickness is regulated independently for each axon, at a site very near the axon.

Animals↗
PubMed1983

Opioid peptide-like immunoreactivity localized in GABAErgic neurons of rat neostriatum and central amygdaloid nucleus.

Opioid peptide-like (OPL)-immunoreactivity and (the GABA-biosynthetic enzyme) glutamic acid decarboxylase-like (GAD)-immunoreactivity were localized in rat neostriatum and central amygdaloid nucleus (ACE) using a polyclonal sheep antiserum to rat brain GAD and a monoclonal mouse antibody to the N-terminus of beta-endorphin (3-E7) as primary antisera. PAP-immunohistochemistry revealed GAD-immunoreactivity in the majority of neurons in neostriatum and ACE. OPL-immunoreactivity was observed in numerous neurons in ACE, but only in few neostriatal nerve cells. In double immunofluorescence in the same section OPL- and GAD-immunoreactivity colocalized in few medium size cells in the neostriatum, but in numerous neurons in ACE. The existence of opioid peptide containing GABAergic neurons in ACE and neostriatum is demonstrated.

Amygdala↗
PubMed1983

Zinc-aldehyde fixation for light-microscopic immunocytochemistry of nervous tissues.

Two procedures are described for vascular perfusion of the nervous system with a zinc-aldehyde fixative. The procedures, simple and economical, combine the advantages of perfusion fixation with an aldehyde solution and matrix stabilization by a mordating agent, and improve the sensitivity of the peroxidase-antiperoxidase (PAP) method for the immunocytochemical localization of several antigens. Procedure A is intended for the light-microscopic immunostaining of cellular elements containing high concentrations of antigen. Penetration of the immunoreagents is adequate without the use of detergents. Procedure B is particularly advantageous for the light-microscopic immunostaining of cellular elements that contain low concentrations of antigen, and for high-resolution microphotography. With procedure B, the tissue penetration of immunoreagents is more limited than with procedure A; however, neuronal cell bodies and dendrites are more easily penetrated by the immunoreagents than are axons. Neuronal cell bodies and dendrites thus become clearly detectable in the light-microscope, even when they are surrounded by numerous immunoreactive axon terminals, and especially after the blockage of axoplasmic transport by the topical injection of colchicine.

Animals↗
PubMed1982

Cell junctions and intramembrane particles of astrocytes and oligodendrocytes: a freeze-fracture study.

The plasma membranes of astrocytes and oligondendrocytes in the white matter of the cat were studied with the freeze-fracturing technique. The intramembrane particle profiles differ in the two type of cell. Orthogonal, small particle assembles and isolated globular particles 5-18 nm in diameter characterize the astrocytic plasmalemma, whereas the plasma membrane of oligodendrocytes shows large, tall globular particles, small globular particles, small ellipsoidal particles and previously undescribed, thin, short, rectilinear strands composed of fused subunits. Using these distinct differential features we can identify partners of glial cell junctions. We confirm the existence of interastrocytic gap junctions. Moreover, we identify numerous heterologous gap junctions between astrocytes and oligodendrocytic cell bodies, processes and the outer turn of myelin sheaths. Interoligodendrocytic gap junctions are not observed. Adjacent oligodendrocytes, however, form tight junctions consisting of linear P face strands and rows of particles; tight junctions are a reliable marker for oligodendroglial membranes. Connexons of interastrocytic gap junctions are packed in a crystalline array, while astrocyte-oligodendrocyte junctional connexons are closely packed but not crystalline. This study indicates that gap junctions between glial cells are pleomorphic and non-randomly distributed. The junctions between astrocytes and those between astrocytes and oligodendrocytes may had different roles in interglial and neuron-glia cooperation.

Animals↗