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BACKGROUND: Rotavirus (RV) infection is the first cause of acute viral gastroenteritis in children under five years of age all over the world; it mainly affects children between six and 24 months of age and can cause serious acute diarrhoea and dehydration. The aim of this study is to perform the budget impact analysis of universal rotavirus vaccination in the Local Health Unit (LHU) 11 Empoli, Tuscany, Italy. METHODS: An ad hoc mathematical simulation model was developed to evaluate the budget impact analysis of 5-years universal rotavirus vaccination. Particularly, incidence of rotavirus gastroenteritis (RVGE), hospitalizations, nosocomial diarrhoea, medical consultations, prescriptions and accesses to emergency department were taken into account in the analysis. The direct medical costs due to RV diarrhoea and the costs of vaccination campaign were considered as the main outcome measures in the study. RESULTS: The adoption of universal rotavirus vaccination campaign for five years in the LHU 11 Empoli would result in relevant savings due to the health cares avoided. These savings would overlapped the costs of vaccination yet from the second year after the introduction of vaccination. The saving for the Health Service would be 1.5 million Euro after five years of campaign. CONCLUSIONS: Universal vaccination against rotavirus results clinically and economically favourable for both the Health Service and the Society perspectives.
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The chick ciliary ganglion receives a nonvascular symathetic innervation in addition to the well-known cholinergic one; fluorescent, varicose adrenergic fibers form pericelluar baskets. Adrenergic fibes were identified electron microscopically in ganglia fixed with potassium permanganate. The fibers degenerate after injection of 6-hydroxydopamine. No true synaptic relationships involving adrenergic varicosities and ganglion cells or cholinergic terminals were demonstrable. The distribution of the adrenergic fibers suggests a kind of "distance à synapse" with the choroidal cells or with the preganglionic fibers (or both). The adrenergic innervation might provide a modulation of the cholinergic transmission.
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Cuerpo Ciliar/inervación , Ganglios/citología , Sistema Nervioso Parasimpático/citología , Sistema Nervioso Simpático/citología , Animales , Pollos , Femenino , Microscopía Electrónica , Microscopía Fluorescente , Neuroglía/citología , Neuronas/citología , Sinapsis/citología , Vesículas SinápticasRESUMEN
We have recorded from the granular layer of the turtle cerebellum extracellular unitary potentials that appear to reflect pre- and postsynaptic events at the synapse between a single swelling of a mossy fiber and the dendritic tips of several granule cells. The presynaptic component is an all-or-none potential. It can be directly activated by spinal stimulation and is unaltered by repetitive activity or by high concentrations of magnesium. The postsynaptic component is a graded potential. It follows the presynaptic component by approximately 1 millisecond and is depressed by repetitive activity and by high concentrations of magnesium. The recording of large potentials produced by the flow of postsynaptic current within a single glomerulus suggests powerful transmission. Electron micrographs demonstrate large cerebellar glomeruli in the turtle and a substantial accumulation of mitochondria in the dendritic tips of granule cells.
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Cerebelo/fisiología , Sinapsis/fisiología , Transmisión Sináptica , Potenciales de Acción , Animales , Cerebelo/citología , Estimulación Eléctrica , Microscopía Electrónica , Mitocondrias , Médula Espinal/fisiología , Vesículas Sinápticas , TortugasRESUMEN
The cerebellum has many properties that make it a useful model for investigating neural development. Purkinje cells, the major output neurons of the cerebellar cortex, have drawn special attention because of the availability of biochemical markers and mutants that affect their development. The spatial expression of L7, a protein specific for Purkinje cells, and L7 beta Gal, a gene expressed in transgenic mice that was constructed from the L7 promoter and the marker beta-galactosidase, delineated bands of Purkinje cells that increased in number during early postnatal development. Expression of the transgene in adult reeler mutant mice, which show inverted cortical lamination, and in primary culture showed that the initial expression of L7 is intrinsic to Purkinje cells and does not depend on extracellular signals. This may reflect an underlying developmental map in cerebellum.
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Cerebelo/crecimiento & desarrollo , Células de Purkinje/fisiología , beta-Galactosidasa/genética , Envejecimiento , Animales , Cerebelo/citología , Cerebelo/embriología , Desarrollo Embrionario y Fetal , Ratones , Ratones Mutantes Neurológicos , Ratones Transgénicos , Células de Purkinje/citología , Proteínas Recombinantes/metabolismo , beta-Galactosidasa/metabolismoRESUMEN
The inositol 1,4,5-trisphosphate receptor (IP3R) is expressed at very high levels in cerebellar Purkinje cells. Within these neurons, it has a widespread distribution throughout the endoplasmic reticulum (ER) and is present at particularly high concentrations at sites of membrane appositions within peculiar stacks of ER cisternae. Here we report that stacks of ER cisternae, reminiscent of those observed in Purkinje cells, can be induced by overexpression of full-length IP3R, but not of mutant forms of the protein in COS cells. Within these stacks the IP3R forms a crystalline array at apposed cisternal faces. Additionally, we show that Purkinje cell stacks are not permanent structures. Our findings suggest that massive stack formation in purkinje cells represents an adaptive response of the ER to hypoxic conditions and is due to the presence of the high concentration of IP3R in its membranes.
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Canales de Calcio/fisiología , Retículo Endoplásmico/fisiología , Fibroblastos/fisiología , Células de Purkinje/fisiología , Receptores Citoplasmáticos y Nucleares/fisiología , Transfección , Animales , Secuencia de Bases , Canales de Calcio/genética , Línea Celular , Retículo Endoplásmico/ultraestructura , Fibroblastos/ultraestructura , Receptores de Inositol 1,4,5-Trifosfato , Masculino , Microscopía Electrónica , Datos de Secuencia Molecular , Sondas de Oligonucleótidos/genética , Reacción en Cadena de la Polimerasa , Células de Purkinje/ultraestructura , Ratas , Ratas Sprague-Dawley , Receptores Citoplasmáticos y Nucleares/genéticaRESUMEN
Axon guidance and target recognition depend on neuronal cell surface receptors that recognize and elicit selective growth cone responses to guidance cues in the environment. Contactin, a cell adhesion/recognition molecule of the immunoglobulin gene superfamily, regulates axon growth and fasciculation in vitro, but its role in vivo is unknown. To assess its function in the developing nervous system, we have ablated contactin gene expression in mice. Contactin-/- mutants displayed a severe ataxic phenotype consistent with defects in the cerebellum and survived only until postnatal day 18. Analysis of the contactin-/- mutant cerebellum revealed defects in granule cell axon guidance and in dendritic projections from granule and Golgi cells. These results demonstrate that contactin controls axonal and dendritic interactions of cerebellar interneurons and contributes to cerebellar microorganization.
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Ataxia/genética , Moléculas de Adhesión Celular Neuronal/metabolismo , Cerebelo/patología , Animales , Animales Recién Nacidos/metabolismo , Ataxia/complicaciones , Ataxia/mortalidad , Ataxia/patología , Moléculas de Adhesión Celular Neuronal/genética , Corteza Cerebelosa/metabolismo , Contactinas , Dendritas/patología , Dendritas/ultraestructura , Aparato de Golgi/ultraestructura , Ratones , Ratones Noqueados/genética , Fibras Nerviosas/patología , Fenotipo , Células de Purkinje/ultraestructuraRESUMEN
In most mammals the cochlear nuclear complex (CN) contains a distributed system of granule cells (GCS), whose parallel fiber axons innervate the dorsal cochlear nucleus (DCN). Like their counterpart in cerebellum, CN granules are innervated by mossy fibers of various origins. The GCS is complemented by unipolar brush (UBCs) and Golgi cells, and by stellate and cartwheel cells of the DCN. This cerebellum-like microcircuit modulates the activity of the DCN's main projection neurons, the pyramidal, giant and tuberculoventral neurons, and is thought to improve auditory performance by integrating acoustic and proprioceptive information. In this paper, we focus on the rat UBCs, a chemically heterogeneous neuronal population, using antibodies to calretinin, metabotropic glutamate receptor 1alpha (mGluR1alpha), epidermal growth factor substrate 8 (Eps8) and the transcription factor T-box gene Tbr2 (Tbr2). Eps8 and Tbr2 labeled most of the CN's UBCs, if not the entire population, while calretinin and mGluR1alpha distinguished two largely separate subsets with overlapping distributions. By double labeling with antibodies to Tbr2 and the alpha6 GABA receptor A (GABAA) subunit, we found that UBCs populate all regions of the GCS and occur at remarkably high densities in the DCN and subpeduncular corner, but rarely in the lamina. Although GCS subregions likely share the same microcircuitry, their dissimilar UBC densities suggest they may be functionally distinct. UBCs and granules are also present in regions previously not included in the GCS, namely the rostrodorsal magnocellular portions of ventral cochlear nucleus, vestibular nerve root, trapezoid body, spinal tract and sensory and principal nuclei of the trigeminal nerve, and cerebellar peduncles. The UBC's dendritic brush receives AMPA- and NMDA-mediated input from an individual mossy fiber, favoring singularity of input, and its axon most likely forms several mossy fiber-like endings that target numerous granule cells and other UBCs, as in the cerebellum. The UBCs therefore, may amplify afferent signals temporally and spatially, synchronizing pools of target neurons.
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Núcleo Coclear/citología , Red Nerviosa/metabolismo , Neuronas/citología , Neuronas/metabolismo , Fenotipo , Proteínas Adaptadoras Transductoras de Señales , Animales , Mapeo Encefálico , Calbindina 2 , Fibras Nerviosas/metabolismo , Proteínas/metabolismo , Ratas , Ratas Sprague-Dawley , Receptores de GABA-A/metabolismo , Receptores de Glutamato Metabotrópico/metabolismo , Proteína G de Unión al Calcio S100/metabolismo , Proteínas de Dominio T Box/metabolismoRESUMEN
Epidermal growth factor receptor pathway substrate 8 (Eps8) is a widely expressed multidomain signaling protein that coordinates two disparate GTPase-dependent mechanisms: actin reorganization via Ras/Rac pathways and receptor trafficking via Rab5. Expression of Eps8, the gene encoding the founding member of the Eps8 family of proteins, was found in cerebellum by virtual Northern analysis and in situ hybridization. Because the cerebellum has a well-known cellular architecture and is a favored model to study synaptic plasticity and actin dynamics, we sought to analyze Eps8 localization in rat cerebellar neurons and synapses by light and electron microscopy. Specificity of Eps8-antibody was demonstrated by immunoblots and in brain sections. In cerebellum, unipolar brush cells (UBCs) were densely Eps8 immunopositive and granule cells were moderately immunostained. In both types of neuron immunoreaction product was localized to the somatodendritic and axonal compartments. Postsynaptic immunostained foci were demonstrated in the glomeruli in correspondence of the synapses formed by mossy fiber terminals with granule cell and UBC dendrites. These foci appeared especially evident in the UBC brush, which contains an extraordinary postsynaptic apparatus of actin microfilaments facing synaptic junctions of the long and segmented varieties. Eps8 immunoreactivity was conspicuously absent in Purkinje cells and their actin-rich dendritic spines, in all types of inhibitory interneurons of the cerebellum, cerebellar nuclei neurons, and astrocytes. In conclusion, Eps8 protein in cerebellum is expressed exclusively by excitatory cortical interneurons and is intracellularly compartmentalized in a cell-class specific manner. This is the first demonstration of the presence of a member of the Eps8 protein family in UBCs and its enrichment at postsynaptic sites.
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Cerebelo/citología , Neuronas/citología , Terminales Presinápticos/metabolismo , Proteínas/metabolismo , Proteínas Adaptadoras Transductoras de Señales , Animales , Animales Recién Nacidos , Expresión Génica/fisiología , Inmunohistoquímica/métodos , Lisina/análogos & derivados , Lisina/metabolismo , Masculino , Microscopía Inmunoelectrónica/métodos , Proteínas del Tejido Nervioso/metabolismo , Neuronas/ultraestructura , Terminales Presinápticos/ultraestructura , Ratas , Ratas Sprague-Dawley , Ratas WistarRESUMEN
Cerebellar morphogenesis occurs through a complex interplay of cell proliferation and migration that in mouse and rat begins about midgestation and ends in the third postnatal week. Cerebellar cells derive from germinative matrices in the ventricular zone and the external granular layer. Like granule cells, unipolar brush cells (UBCs) are excitatory interneurons situated in the granular layer of the cortex and innervated by mossy fibers. While granule cells are produced from the external granular layer, the generation of UBCs is still controversial. We utilized the reeler mutant mouse, which has widespread misplacement of neurons due to lack of Reelin protein, to ascertain the origin of UBCs. In the reeler cerebellum, which is small and lacks foliation, Purkinje cells are greatly reduced in number and in large part are located ectopically in deep cerebellar masses. Granule cells are also reduced in number and form an irregular granule cell layer. In this study we demonstrate that the reeler mutation influences the positioning of UBCs and also significantly reduces their number. Both subsets of UBCs identified in normal mouse, the calretinin-positive and the metabotropic glutamate receptor 1alpha-positive subsets, are affected in the reeler. About 40% of the calretinin-positive UBCs are ectopically situated in the deep cerebellar regions and the immediate vicinity of the ependyma of the fourth ventricle. Ectopic UBCs have discrete, although somewhat looser brushes than granular layer UBCs, but form synaptic junctions with complex axon terminals, possibly belonging to mossy fibers and UBC axons, like their normally situated counterpart. The observed displacement of UBCs in the reeler suggests that they originate from the ventricular zone.
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Cerebelo/patología , Interneuronas/fisiología , Ratones Mutantes Neurológicos/anatomía & histología , Animales , Mapeo Encefálico , Calbindina 2 , Recuento de Células/métodos , Cerebelo/anomalías , Cerebelo/metabolismo , Inmunohistoquímica/métodos , Interneuronas/ultraestructura , Ratones , Microscopía Electrónica de Transmisión/métodos , Proteína Reelina , Proteína G de Unión al Calcio S100/metabolismo , Técnicas Estereotáxicas , Sinaptofisina/metabolismoRESUMEN
OBJECTIVE: To compare the T-cell receptor (TCR) repertoire of T-cell subsets in peripheral blood and lymphoid tissue from HIV-1 infected individuals. DESIGN: Biopsies of tonsillar tissue and samples of peripheral blood were obtained from 10, mostly treatment-naive, HIV-1-infected individuals. CD4 and CD8 T-cell subsets were quantified, the TCR repertoire was analysed within 'naive' and 'memory' subsets, and results compared between identical subsets in tonsillar tissue and blood. METHODS: Cell subsets were quantified by flow cytometry. CD4 T cells and CD8 T cells were isolated by immunomagnetic beads. Populations were in most cases further subdivided by immunomagnetic selection on the basis of CD45RO expression. TCR repertoire was studied by spectratyping of the TCR beta variable (BV) complementarity determining region 3 (CDR3) transcripts. RESULTS: Amongst CD4 T cells, an abnormal TCR repertoire was found in median 25% (range, 0-88%) of BV families in peripheral blood, but in 0% (0-7%) in tonsillar tissue (P<0.05). Large peaks suggestive of expanded clones were common within CD8 T-cells, both in peripheral blood and tonsillar tissue. However, the expanded clones were rarely identical in the two compartments. Expanded CDR3 peaks, suggesting the presence of clonally expanded cells, were observed within both CD45RO+ and CD45RO- cells from all T-cell subsets, but, again they were mainly of different lengths. CONCLUSION: CD4 T cells were preserved in number and TCR repertoire in tonsillar tissue compared with blood in HIV-1 infected individuals. T-cells collected from the peripheral blood may not be representative of those residing in lymphoid tissue.
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Antígenos CD4/metabolismo , Regiones Determinantes de Complementariedad , Infecciones por VIH/inmunología , Tonsila Palatina/inmunología , Adulto , Linfocitos B/inmunología , Antígenos CD4/sangre , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/inmunología , Recuento de Células , Citometría de Flujo , Infecciones por VIH/sangre , Humanos , Región Variable de Inmunoglobulina/inmunología , Receptores de Antígenos de Linfocitos T/sangre , Receptores de Antígenos de Linfocitos T/metabolismoRESUMEN
A series of overlapping clones coding for L-glutamic acid decarboxylase was purified from a mouse brain cDNA library, the longest of which contains a 1869 bp open reading frame and 913 bp of non-coding sequence. By comparison with the corresponding sequences from the mouse genome, it was determined that the first methionine in the longest cDNA represents the initiation codon. Expression of this cDNA in eukaryotic cells produces a 62 kd protein that is recognized by antiserum against rat GAD and which displays GAD activity commensurate with the amount of protein produced. Antibodies raised against the purified product of this cDNA recognize a 62 kd protein from mouse brain on immunoblots, specifically stain GABA-ergic neurons in brain sections, and are capable of immunoprecipitating most GAD activity from mouse brain extracts. These results provide the first definitive identification of a cDNA coding for the larger of two forms of GAD in mouse brain, and suggest that the two forms are closely related.
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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.
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Nervio Coclear/anatomía & histología , Ácido gamma-Aminobutírico/metabolismo , Animales , Axones/ultraestructura , Cerebelo/anatomía & histología , Dendritas/ultraestructura , Glutamato Descarboxilasa/metabolismo , Técnicas para Inmunoenzimas , Red Nerviosa/anatomía & histología , Neuronas/clasificación , Neuronas/ultraestructura , Ratas , Ratas Endogámicas , Sinapsis/ultraestructura , Membranas Sinápticas/ultraestructuraRESUMEN
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)
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Cerebelo/anatomía & histología , Pollos/anatomía & histología , Macaca mulatta/anatomía & histología , Macaca/anatomía & histología , Fibras Nerviosas/anatomía & histología , Animales , Corteza Cerebelosa/anatomía & histología , Femenino , Degeneración NerviosaRESUMEN
Electric fish generate an electric field, which they sense with cutaneous electroreceptors. Electroreceptors project topographically onto the medullary electrosensory lateral line lobe (ELL). The ELL of gymnotiform electric fish is divided into four segments specialized to detect different aspects of the electrosensory input; it is also laminated with separate laminae devoted to electroreceptive input, interneurons, projection neurons, and feedback input. We have utilized antisera to glutamic acid decarboxylase (GAD) and gamma-aminobutyric acid (GABA) to map the distribution of GABAergic cells and fibers in the ELL of the gymnotiform fish, Apteronotus leptorhynchus. Six types of GABAergic interneurons are found in ELL: Type 2 granular cells (granular layer) project to pyramidal cells; polymorphic cells (pyramidal cell layer) project to the non-GABAergic type 1 granular cells; ovoid cells (deep neuropil layer) project bilaterally upon basilar dendrites of pyramidal cells; multipolar cells (deep neuropil layer) project bilaterally, probably to dendrites and neurons within the deep neuropil layer; and neurons of the ventral molecular layer and stellate cells (molecular layer) project to apical dendrites of pyramidal cells. GABAergic bipolar cells in the nucleus praeminentialis, a rhombencephalic structure devoted to feedback in the electrosensory system, project in relatively diffuse fashion to pyramidal cells. We hypothesize that the various GABAergic circuits of the ELL can be correlated with specific functions: type 2 granular cells with adaptation, size of receptive field center, and gain; polymorphic cells and type 1 granular cells with regulation of surround inhibition; ovoid cells with common mode rejection; and neurons of the ventral molecular layer with adaptive gain control. The feedback GABAergic input from bipolar cells of n. praeminentialis to pyramidal cells may be part of a searchlight mechanism similar to the one postulated for thalamocortical systems.
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Pez Eléctrico/fisiología , Neuronas Aferentes/fisiología , Órganos de los Sentidos/fisiología , Ácido gamma-Aminobutírico/fisiología , Animales , Dendritas/metabolismo , Glutamato Descarboxilasa/inmunología , Glutamato Descarboxilasa/metabolismo , Peroxidasa de Rábano Silvestre , Inmunohistoquímica , Interneuronas/fisiología , Mecanorreceptores/fisiología , Vías Nerviosas/citología , Vías Nerviosas/fisiología , Neuronas Aferentes/ultraestructura , Células Piramidales/fisiología , Órganos de los Sentidos/citología , Canales de Sodio/metabolismo , Adhesión del Tejido , Aglutinina del Germen de Trigo-Peroxidasa de Rábano Silvestre Conjugada , Aglutininas del Germen de Trigo , Ácido gamma-Aminobutírico/inmunología , Ácido gamma-Aminobutírico/metabolismoRESUMEN
We describe with a variant of the Golgi method a new type of neuron that is prominently represented in the granular layer of the mammalian vestibulocerebellum but is presently neglected in all major accounts on the cerebellum. These neurons, here termed unipolar brush cells, are intermediate in size between granule cells and Golgi cells. They typically have a thin and presumably myelinated axon, and a single and stubby dendrite whose tip forms a tightly packed group of branchlets resembling a paintbrush. The branchlets often intertwine with the digitiform claws of granule cell dendrites and are occasionally approached by Golgi cell dendrites, indicating that the unipolar brush cells may share the input of the other granular layer neurons. Branchlets of neighboring unipolar brush cells converging into the same neuropil island also occur. The brush-like tip of the unipolar cell engulfs one or two mossy fiber rosettes to form an extensive synapse that appears to close recurrent loops involving the vestibular nuclei. Positive feedback in these loops could help to explain several motor responses and drive mechanisms of extended duration that are controlled by the ventral cerebellum.
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Corteza Cerebelosa/ultraestructura , Neuronas/ultraestructura , Animales , Gatos , Corteza Cerebelosa/fisiología , Dendritas/ultraestructura , Retroalimentación/fisiología , Histocitoquímica , Macaca mulatta , Ratones , Ratones Endogámicos C57BL , Fibras Nerviosas/ultraestructura , Neuronas/fisiología , Ratas , Ratas Sprague-Dawley , Sinapsis/ultraestructura , Núcleos Vestibulares/fisiología , Núcleos Vestibulares/ultraestructuraRESUMEN
The projection of the vestibular nerve to the cerebellum of the cat was examined with silver degeneration methods after complete lesions of the vestibular ganglion. The majority of the primary vestibular afferents were traced to the cortex of the ipsilateral nodulus and uvula, relatively fewer entering the ipsilateral flocculus. Fibers were not traced to the paraflocculus, lingula or lateral cerebellar nucleus. A sparse projection to the ipsilateral fastigial nucleus may exist, but it remains equivocal until confirmed with additional methods. Light microscopic examination of plastic sections confirmed these observations and showed further details of the organization of the primary vestibular projection to the nodulus and uvula. These results show that the region of the cerebellum densely innervated by primary vestibular afferents is smaller than previously believed.
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Cerebelo/anatomía & histología , Nervio Vestibular/anatomía & histología , Vías Aferentes/anatomía & histología , Animales , Mapeo Encefálico , Gatos , Corteza Cerebelosa/anatomía & histología , Núcleos Cerebelosos/anatomía & histologíaRESUMEN
The cochlear nuclear complex of the cat was immunostained with an antiserum to glutamate decarboxylase (GAD), the biosynthetic enzyme for the inhibitory neurotransmitter GABA, and studied with different procedures, including silver intensification, topical colchicine injections, semithin sections, and immunoelectron microscopy. Immunostaining was found in all portions of the nucleus. Relatively few immunostained cell bodies were observed: most of these were in the dorsal cochlear nucleus and included stellate cells, cartwheel cells, Golgi cells, and unidentified cells in the deep layers. An accumulation of immunoreactive cells was also found within the small cell cap and along the medial border of the ventral cochlear nucleus. Immunostained cells were sparse in magnocellular portions of the ventral nucleus. Most staining within the nucleus was of nerve terminals. These included small boutons that were prominent in the neuropil of the dorsal cochlear nucleus, the granule cell domain, in a region beneath the superficial granule cell layer within the small cell cap region, and along the medial border of the ventral nucleus. Octopus cells showed small, GAD-positive terminals distributed at moderate density on both cell bodies and dendrites. Larger, more distinctive terminals were identified on the large cells in the ventral nucleus, in particular on spherical cells and globular cells. There was a striking positive correlation of the size, location, and complexity of GAD-positive terminals with the size, location, and complexity of primary fiber endings on the same cells. This correlation did not hold in the dorsal nucleus, where pyramidal cells receive many large GAD-positive somatic terminals despite the paucity of primary endings on their cell bodies. The GAD-positive terminals contained pleomorphic synaptic vesicles and formed symmetric synaptic junctions that occupied a substantial portion of the appositional surface to cell bodies, dendrites, axon hillocks, and the beginning portion of the initial axon segments. Thus, the cells provided with large terminals can be subjected to considerable inhibition that may be activated indirectly through primary fibers and interneurons or by descending inputs from the auditory brainstem.
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Gatos/metabolismo , Nervio Coclear/enzimología , Glutamato Descarboxilasa/metabolismo , Animales , Nervio Coclear/citología , Nervio Coclear/ultraestructura , Aumento de la Imagen , Inmunohistoquímica , Microscopía Electrónica , Terminaciones Nerviosas/enzimología , Neuronas/clasificación , Plata , Coloración y EtiquetadoRESUMEN
The unipolar brush cells (UBCs), a class of neurons recently identified in the granular layer of the vestibulocerebellum, receive excitatory synaptic input from mossy fibers (MFs) in the form of a giant glutamatergic synapse. UBCs are provided with axons that bear synaptic endings situated at the center of glomeruli, similar to cerebellar MF afferents. A single MF stimulus evokes a prolonged train of action potentials in the UBC (Rossi et al., 1995), which is presumably distributed to postsynaptic targets. Knowledge of the synaptic connections of UBC axons is essential to define the role of these cells in the integration of vestibular signals in the cerebellar circuitry. To evaluate these connections, the nodulus (folium X) was isolated from vermal slices of postnatal day 8 mice, cultured for 2-4 or 15-30 days in vitro, and studied by electron and fluorescence microscopy. The peak of degeneration of extrinsic MF terminals, which have been severed from the parent cell bodies, was observed at 2 days in vitro (DIV). Quantification of degenerating and nondegenerating (e.g., intrinsic) MF terminals indicated that about half of the MF terminals were provided by local UBC axons synapsing on dendrites of granule cells and other UBCs. The proportion of nondegenerating vs. degenerating MF terminals terminating on UBCs also indicated that approximately two-thirds of the intrinsic MFs are involved in UBC-UBC connections. In long-term cultures, the granular layer appeared well preserved and the UBC axons formed an extensive system of MF collaterals. It is suggested that UBCs may act by spatially amplifying vestibular inputs carried by extrinsic MFs.
Asunto(s)
Axones/fisiología , Cerebelo/fisiología , Fibras Nerviosas/fisiología , Sinapsis/fisiología , Animales , Animales Recién Nacidos , Axones/ultraestructura , Células Cultivadas , Cerebelo/ultraestructura , Ratones , Ratones Endogámicos C57BL , Fibras Nerviosas/ultraestructura , Sinapsis/ultraestructuraRESUMEN
To ascertain whether the auditory neocortex also innervates the central nucleus of the inferior colliculus (CNIC) and not only its dorsal (DCIC) and external (ECIC) cortices, the anterograde tracers Phaseolus vulgaris-leucoagglutinin (PHA-L) and biotinylated dextran (BD) were injected into the primary auditory neocortex of albino rats (Te1), and labeled corticocollicular fibers were studied via light and electron microscopy. Axons from discrete regions of Te1 form two rostrocaudally oriented laminar plexuses of terminal fibers in the ipsilateral inferior colliculus (IC) and one in the contralateral IC. The first ipsilateral plexus, located in the medial half of the IC, has a dorsomedial to ventrolateral orientation, parallel to the isofrequency planes of the IC; is continuous through the CNIC and DCIC; and extends into the rostral ECIC. The second plexus is located in the deep layers of the lateral ECIC. These two plexuses meet caudally and ventrally, at the border between the CNIC and the lateral ECIC. The plexus in the contralateral IC is less dense and shorter than the two ipsilateral plexuses and is symmetric to the medial plexus. The thickness of the three plexuses is correlated with the size of the injection site, and their mediolateral and dorsoventral positions change as the injection site in Te1 is displaced rostrocaudally, with more caudal injections resulting in more dorsolateral medial plexuses and more dorsomedial lateral plexuses. Furthermore, the ventromedial border of the IC receives nontopographic, convergent projections from wide regions of rostral portions of Te1. The distribution of these corticocollicular plexuses mimics the topography of previously described intracollicular fibers. Electron microscopy shows that, in all three subdivisions of the ipsilateral IC, corticocollicular fibers form small boutons with features generally associated with excitatory transmission; i.e., they contain round synaptic vesicles and form asymmetric synapses with thin dendritic shafts and spines. These results demonstrate that the auditory corticocollicular projections innervate more extensive regions of the IC than were previously observed. Although peripheral regions receive the densest projection, the entire IC appears to be the target of corticofugal input.
Asunto(s)
Corteza Auditiva/anatomía & histología , Mapeo Encefálico , Colículos Inferiores/anatomía & histología , Animales , Núcleo Coclear/anatomía & histología , Vías Eferentes/anatomía & histología , Femenino , Microscopía Electrónica , RatasRESUMEN
The cerebellar cortex of adult hens contains a dense plexus of thin varicose nerve fibers which display a formaldehyde-induced green fluorescence. This plexus is not distributed at random in the cortical layers. Within the granular layer the plexus forms a netlike pattern. The fiber branches, which have numerous varicosities, are predominantly oriented in the traverse plane of the folium. In the molecular layer the fluorescent plexus shows some variations in the convex, flat and concave portions of the folia. Many of the fluorescent branches are oriented parallel to the course of the folium. They arise from a T-division of radially oriented axons resembling parallel fibers in Golgi sections. The meshes of the fluorescent plexus in the granular layer measure 10-60 mu. In the molecular layer (top of the folia) there are about 30 fluorescent fibers per 100 mu2. The fluorescent fibers originate from the locus coeruleus and form a rostral and a caudal bundle in the cerebellar peduncle. The mode of distribution of the fluorescent fibers in the cortical layers seems to depend on the organization of the innervated tissue. Light microscopy suggests that the aminergic fibers innervate more than one class of cerebellar neurons.