RESUMEN
In the post-natal rodent brain, neuronal precursors originating from the sub-ventricular zone (SVZ) migrate over a long distance along the rostral migratory stream (RMS) to eventually integrate the olfactory bulb neuronal circuitry. In order to identify new genes specifically expressed in the RMS, we have screened the Allen Brain Atlas Database. We focused our attention on Thrombospondin 4 (Thbs4), one of the 5 members of the Thrombospondin family of large, multidomain, extracellular matrix proteins. In post-natal and adult brain Thbs4 mRNA and protein are specifically expressed in the neurogenic regions, including the SVZ and along the entire RMS. RMS cells expressing Thbs4 are GFAP (Glial Fibrillary Acidic Protein) positive astrocytes. Histological analysis in both wild-type and Thbs4 knock-out mice revealed no major abnormality in the general morphology of these neurogenic regions. Nevertheless, immunostaining for doublecortin demonstrates that in Thbs4-KO, migration of newly formed neurons along the RMS is somehow impaired, with several neurons migrating out of the RMS. This is further supported by a Bromodeoxyuridine-based in vivo approach showing a decrease in the number of newly born neuronal precursors reaching the olfactory bulb, while proliferation in the SVZ is not affected compared to wild-type, both in young animals (P15) and in adults (8 to 12 weeks of age). Corroborating this observation, the number of Parvalbumin- and Calbindin-immunoreactive interneurons in the olfactory bulb is also reduced in Thbs4-KO. Together, these observations support a role for the astrocyte-secreted protein Thbs4 in the migration of newly form neurons within the RMS to the olfactory bulb.
Asunto(s)
Envejecimiento , Movimiento Celular/genética , Regulación del Desarrollo de la Expresión Génica/genética , Ventrículos Laterales/citología , Ventrículos Laterales/crecimiento & desarrollo , Trombospondinas/deficiencia , Animales , Animales Recién Nacidos , Astrocitos/metabolismo , Proliferación Celular/genética , Proteína Ácida Fibrilar de la Glía/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Vías Nerviosas/fisiología , ARN Mensajero/metabolismo , Trombospondinas/genéticaRESUMEN
gamma-Aminobutyric acid (GABA) is one of the major inhibitory neurotransmitters in the central nervous system. In the cerebral cortex, GABA-containing cells represent a subpopulation of interneurons. With semithin frozen sections, it is possible to demonstrate that most GABA neurons in the rat somatosensory cortex contain the calcium-binding protein parvalbumin and that parvalbumin is found virtually only in GABA neurons. Parvalbumin seems to influence the electrical properties and enzymatic machinery to modulate neuronal excitability and activity. The specific role of parvalbumin in GABA-containing cortical cells may be related to controlling the effectiveness of their inhibitory action.
Asunto(s)
Corteza Cerebral/citología , Proteínas Musculares/análisis , Neuronas/análisis , Parvalbúminas/análisis , Ácido gamma-Aminobutírico/fisiología , Animales , Corteza Cerebral/análisis , Electrofisiología , Masculino , Neuronas/fisiología , RatasRESUMEN
Rapid eye movements (REM) are characteristic of the eponymous phase of sleep, yet the underlying motor commands remain an enigma. Here, we identified a cluster of Calbindin-D28K-expressing neurons in the Nucleus papilio (NPCalb), located in the dorsal paragigantocellular nucleus, which are active during REM sleep and project to the three contralateral eye-muscle nuclei. The firing of opto-tagged NPCalb neurons is augmented prior to the onset of eye movements during REM sleep. Optogenetic activation of NPCalb neurons triggers eye movements selectively during REM sleep, while their genetic ablation or optogenetic silencing suppresses them. None of these perturbations led to a change in the duration of REM sleep episodes. Our study provides the first evidence for a brainstem premotor command contributing to the control of eye movements selectively during REM sleep in the mammalian brain.
Asunto(s)
Movimientos Oculares/fisiología , Bulbo Raquídeo/fisiología , Neuronas Motoras/fisiología , Neuronas/fisiología , Animales , Electroencefalografía , Electromiografía , Electrooculografía , Humanos , Macaca fascicularis , Macaca mulatta , Bulbo Raquídeo/citología , Ratones Endogámicos C57BL , Ratones Transgénicos , OptogenéticaRESUMEN
AIMS: Prion diseases are generally characterized by pronounced neuronal loss. In particular, a subpopulation of inhibitory neurones, characterized by the expression of the calcium-binding protein parvalbumin (PV), is selectively destroyed early in the course of human and experimental prion diseases. By contrast, nerve cells expressing calbindin D28 k (CB), another calcium-binding protein, as well as PV/CB coexpressing Purkinje cells, are well preserved. METHODS: To evaluate, if PV and CB may directly contribute to neuronal vulnerability or resistance against nerve cell death, respectively, we inoculated PV- and CB-deficient mice, and corresponding controls, with 139A scrapie and compared them with regard to incubation times and histological lesion profiles. RESULTS: While survival times were slightly but significantly diminished in CB-/-, but not PV-/- mice, scrapie lesion profiles did not differ between knockout mice and controls. There was a highly significant and selective loss of isolectin B(4)-decorated perineuronal nets (which specifically demarcate the extracellular matrix surrounding the 'PV-expressing' subpopulation of cortical interneurones) in scrapie inoculated PV+/+, as well as PV-/- mice. Purkinje cell numbers were not different in CB+/+ and CB-/- mice. CONCLUSIONS: Our results suggest that PV expression is a surrogate marker for neurones highly vulnerable in prion diseases, but that the death of these neurones is unrelated to PV expression and thus based on a still unknown pathomechanism. Further studies including the inoculation of mice ectopically (over)expressing CB are necessary to determine whether the shortened survival of CB-/- mice is indeed due to a neuroprotective effect of this molecule.
Asunto(s)
Parvalbúminas/deficiencia , Parvalbúminas/metabolismo , Proteína G de Unión al Calcio S100/genética , Proteína G de Unión al Calcio S100/metabolismo , Scrapie/metabolismo , Animales , Calbindina 1 , Calbindinas , Modelos Animales de Enfermedad , Inmunohistoquímica , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Red Nerviosa/patología , Scrapie/genética , Scrapie/patología , Especificidad de la Especie , Análisis de Supervivencia , Vacuolas/patología , Vacuolas/ultraestructuraRESUMEN
Among the many calcium-binding proteins in the nervous system, parvalbumin, calbindin-D28K and calretinin are particularly striking in their abundance and in the specificity of their distribution. They can be found in different subsets of neurons in many brain regions. Although it is not yet known whether they play a 'triggering' role like calmodulin, or merely act as buffers to modulate cytosolic calcium transients, they are valuable markers of neuronal subpopulations for anatomical and developmental studies.
Asunto(s)
Proteínas de Unión al Calcio/metabolismo , Calcio/metabolismo , Proteínas del Tejido Nervioso/metabolismo , Potenciales de Acción , Secuencia de Aminoácidos , Animales , Biomarcadores , Química Encefálica , Canales de Calcio/metabolismo , Proteínas de Unión al Calcio/fisiología , Secuencia de Consenso , Activación del Canal Iónico , Mamíferos/metabolismo , Modelos Moleculares , Datos de Secuencia Molecular , Familia de Multigenes , Degeneración Nerviosa , Proteínas del Tejido Nervioso/fisiología , Neuronas/metabolismo , Neurotransmisores/metabolismo , Retina/metabolismo , Especificidad de la EspecieRESUMEN
Golgi ranked the peripheral reticulum--which adheres intimately to nerve cell surfaces--alongside the intracellular reticulum, or Golgi apparatus,which immortalized his name. At first dismissed as an artefact of capricious staining techniques, this peripheral reticulum, or perineuronal net, is now recognized as a genuine entity in neurocytology. It represents a complex of extracellular matrix molecules interposed between the meshwork of glial processes, from which they are indistinguishable, and nerve-cell surfaces. In no other branch of neuroscience has the waxing and waning of interest in any morphological entity been so pronounced as in the case of the perineuronal net. This review traces the history of this enigmatic structure from its conception to the present time, brings to light the keen observational powers of morphologists at the turn of the century and reveals how their sagacious forethought anticipated current thinking on the role of perineuronal nets.
Asunto(s)
Matriz Extracelular/fisiología , Neuronas/citología , Animales , Encéfalo/citología , Membrana Celular/fisiología , Histocitoquímica/historia , Historia del Siglo XIX , Historia del Siglo XX , Humanos , Neuroglía/citología , Médula Espinal/citologíaRESUMEN
The Ca(2+)-binding proteins parvalbumin (PV) and calbindin D-28k (CB) are key players in the intracellular Ca(2+)-buffering in specific cells including neurons and have profound effects on spatiotemporal aspects of Ca(2+) transients. The previously observed increase in mitochondrial volume density in fast-twitch muscle of PV-/- mice is viewed as a specific compensation mechanism to maintain Ca(2+) homeostasis. Since cerebellar Purkinje cells (PC) are characterized by high expression levels of the Ca(2+) buffers PV and CB, the question was raised, whether homeostatic mechanisms are induced in PC lacking these buffers. Mitochondrial volume density, i.e. relative mitochondrial mass was increased by 40% in the soma of PV-/- PC. Upregulation of mitochondrial volume density was not homogenous throughout the soma, but was selectively restricted to a peripheral region of 1.5 microm width underneath the plasma membrane. Accompanied was a decreased surface of subplasmalemmal smooth endoplasmic reticulum (sPL-sER) in a shell of 0.5 microm thickness underneath the plasma membrane. These alterations were specific for the absence of the "slow-onset" buffer PV, since in CB-/- mice neither changes in peripheral mitochondria nor in sPL-sER were observed. This implicates that the morphological alterations are aimed to specifically substitute the function of the slow buffer PV. We propose a novel concept that homeostatic mechanisms of components involved in Ca(2+) homeostasis do not always occur at the level of similar or closely related molecules. Rather the cell attempts to restore spatiotemporal aspects of Ca(2+) signals prevailing in the undisturbed (wildtype) situation by subtly fine tuning existing components involved in the regulation of Ca(2+) fluxes.
Asunto(s)
Retículo Endoplásmico/patología , Retículo Endoplásmico/ultraestructura , Mitocondrias/patología , Parvalbúminas/deficiencia , Células de Purkinje/ultraestructura , Animales , Western Blotting/métodos , Calbindinas , Calcio/metabolismo , Corteza Cerebelosa/citología , Electroforesis en Gel Bidimensional/métodos , Técnica del Anticuerpo Fluorescente/métodos , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Microscopía Electrónica de Transmisión/métodos , Mitocondrias/ultraestructura , ATPasas Transportadoras de Calcio de la Membrana Plasmática/metabolismo , Células de Purkinje/patología , Proteína G de Unión al Calcio S100/genética , Estadísticas no ParamétricasRESUMEN
Calretinin is a Ca(2+)-binding protein of the EF-hand family which is expressed in colon adenocarcinomas and colon-derived tumor cell lines (e.g. WiDr), but is absent from normal human enterocytes. Its function has not as yet been elucidated, but some lines of evidence lead us to postulate its involvement in cell proliferation in these cells. In order to test whether calretinin is correlated with an undifferentiated, proliferating, or with a differentiated, state of cells, its expression was studied in the human colon adenocarcinoma clonal cell line HT29-18, which can be caused to differentiate into enterocyte-like cells by replacing glucose with galactose in the culture medium (glucose starvation differentiation). Treatment of HT29-18 cells with galactose led to a drop in the calretinin mRNA level and in protein expression as evidenced by immunocytochemical staining and Western blot analysis of cytosolic cell extracts. These results suggest that calretinin is present in HT29-18 cancer cells, mostly in those which are in the undifferentiated state. The possibility that calretinin is involved in maintaining the cells in an undifferentiated (cancerous) state is discussed.
Asunto(s)
Células HT29/citología , Proteína G de Unión al Calcio S100/análisis , Aminopeptidasas/análisis , Calbindina 2 , Muerte Celular , Diferenciación Celular , División Celular , ADN de Neoplasias/análisis , Galactosa , Glucosa , Células HT29/química , Células HT29/enzimología , Células HT29/ultraestructura , Humanos , Microvellosidades/ultraestructura , Índice Mitótico , Proteínas de Neoplasias/análisis , ARN Mensajero/análisis , Proteína G de Unión al Calcio S100/fisiologíaRESUMEN
In mice, 249 putative members of the superfamily of EF-hand domain Ca(2+)-binding proteins, manifesting great diversity in structure, cellular localization and functions have been identified. Three members in particular, namely, calbindin-D28K, calretinin and parvalbumin, are widely used as markers for specific neuronal subpopulations in different regions of the brain. The aim of the present study was to compile a comprehensive atlas of the gene-expression profiles of the entire EF-hand gene superfamily in the murine brain. This was achieved by a meticulous examination of the in-situ hybridization images in the Allen Brain Atlas database. Topographically, our analysis focused on the olfactory bulb, cerebral cortex (barrel cortex in the primary somatosensory area), basal ganglia, hippocampus, amygdala, thalamus, hypothalamus, cerebellum, midbrain, pons and medulla, and on clearly identifiable sub-structures within each of these areas. The expression profiles of four family-members, namely hippocalcin-like 4, neurocalcin-δ, plastin 3 and tescalcin, that have not been hitherto reported, at either the mRNA (in-situ-hybridization) or the protein (immunohistochemical) levels, are now presented for the first time. The fruit of our analysis is a document in which the gene-expression profiles of all members of the EF-hand family genes are compared, and in which future possible neuronal markers for specific cells/brain areas are identified. The assembled information could afford functional clues to investigators, conducive to further experimental pursuit.
Asunto(s)
Encéfalo/metabolismo , Proteínas de Unión al Calcio/genética , Expresión Génica/genética , Estudio de Asociación del Genoma Completo , Neuronas/metabolismo , Envejecimiento , Animales , Proteínas Portadoras/genética , Proteínas Portadoras/metabolismo , Estudio de Asociación del Genoma Completo/métodos , Ratones , ARN Mensajero/metabolismoRESUMEN
We have purified recombinant human calretinin (CR) from Escherichia coli lysates and have produced a polyclonal antiserum against it. The antiserum recognizes determinants conserved in fish, chicken, rat, monkey and human CR. We show its use in the qualitative detection of CR by different methods of immunohistochemistry as well as in the detection of CR on immunoblots.
Asunto(s)
Anticuerpos , Química Encefálica , Proteína G de Unión al Calcio S100/inmunología , Secuencia de Aminoácidos , Animales , Especificidad de Anticuerpos , Secuencia de Bases , Calbindina 2 , Pollos , Escherichia coli , Peces , Haplorrinos , Humanos , Datos de Secuencia Molecular , Ratas , Proteína G de Unión al Calcio S100/química , Proteína G de Unión al Calcio S100/genética , Proteína G de Unión al Calcio S100/aislamiento & purificaciónRESUMEN
We have produced twelve monoclonal antibodies (McAB) against carp-II parvalbumin. Three of them, designated 235, 239, 267 recognize determinants conserved in fish, chicken, mouse, rat, monkey and human parvalbumin. We show their use in the qualitative detection of parvalbumin (PV) by immunohistochemistry, in the quantitation of parvalbumin by radioimmunoassay and in the detection of parvalbumin on immunoblots.
Asunto(s)
Anticuerpos Monoclonales/inmunología , Proteínas de Unión al Calcio/metabolismo , Proteínas Musculares/inmunología , Parvalbúminas/inmunología , Animales , Carpas , Parvalbúminas/metabolismo , Parvalbúminas/fisiologíaRESUMEN
We have produced 25 clones secreting antibodies directed against chicken Calbindin D-28k. Two of them, 300 and 318, recognize determinants conserved in fish, chicken, mouse, rat, rabbit, monkey and human Calbindin D-28k. We demonstrate their use in the immunohistochemical localization of Calbindin D-28k, and in the detection of Calbindin D-28k on immunoblots.
Asunto(s)
Anticuerpos Monoclonales/inmunología , Proteína G de Unión al Calcio S100/inmunología , Animales , Unión Competitiva , Calbindinas , Pollos , Reacciones Cruzadas , Humanos , Immunoblotting , Inmunohistoquímica , Ratas , Proteína G de Unión al Calcio S100/análisis , Proteína G de Unión al Calcio S100/metabolismoRESUMEN
Recent studies have stressed the fact that specific neuronal subtypes may display a differential sensitivity to degeneration in Alzheimer's disease. For example, large pyramidal neurons have been shown to be vulnerable, whereas smaller neurons are resistant to pathology. Using a monoclonal antibody against the calcium-binding protein parvalbumin, we investigated the possible changes in a subpopulation of interneurons in two cortical areas known to be strongly damaged in Alzheimer's disease. In the prefrontal cortex as well as in the inferior temporal cortex, we observed no differences in parvalbumin-immunoreactive cell counts or cell size in Alzheimer's disease brains as compared to control cases. Moreover, the general cellular morphology of these neurons was preserved in the Alzheimer's disease cases, in that their perikarya and dendritic arborizations were intact. These results suggest that paravalbumin-immunoreactive cells represent a neuronal subset resistant to degeneration, and further support the hypothesis that the pathological process in Alzheimer's disease involves specific neuronal subtypes with particular morphological and molecular characteristics.
Asunto(s)
Enfermedad de Alzheimer/patología , Encéfalo/patología , Neuronas/patología , Parvalbúminas/metabolismo , Anciano , Anciano de 80 o más Años , Enfermedad de Alzheimer/metabolismo , Encéfalo/metabolismo , HumanosRESUMEN
Selective neuronal vulnerability and aberrant axonal reorganization in the hippocampus may play an important role for the pathogenesis of pharmaco-resistant temporal lobe epilepsy (TLE). Interneurons containing calcium-binding proteins (CaBPs) are candidates for pathogenetically relevant neurons in the hippocampus of patients with TLE. Here we have examined the cellular localization and distribution of calretinin (CR), a recently discovered CaBP, in the hippocampus of 35 patients with TLE. There was a striking preservation of CR-immunoreactive neurons in TLE patients with Ammon's horn sclerosis (AHS). No significant differences in the distribution of CR-immunoreactive neurons were observed between patients with lesion-associated TLE and control patients without epilepsy. However, a subpopulation of CR-immunoreactive interneurons with morphological features of Cajal-Retzius-like cells, which are only transiently detectable in the normally developing hippocampus, was markedly increased in epilepsy patients with AHS. This increase did not correlate with the duration of the epileptic disorder. Another significant finding was a striking increase and reorganization of CR-immunoreactive neuropil throughout the entire molecular layer of the dentate gyrus (DG-ML) in patients with AHS as compared to patients with focal lesions and control specimens. Ultrastructural analysis identified the CR-immunoreactive axonal profiles as components of an inhibitory, intrinsic neuronal system. The presence of a CR-positive, aberrant cell population, in combination with sprouting of CR-positive axonal processes may significantly alter the gating function of the dentate gyrus and thereby increase hippocampal epileptogenicity in epilepsy patients with AHS.
Asunto(s)
Epilepsia del Lóbulo Temporal/patología , Hipocampo/patología , Neuronas/patología , Proteína G de Unión al Calcio S100/inmunología , Adolescente , Adulto , Edad de Inicio , Anciano , Calbindina 2 , Recuento de Células , Niño , Preescolar , Femenino , Humanos , Inmunohistoquímica , Lactante , Masculino , Persona de Mediana Edad , Factores de TiempoRESUMEN
The existence of angiotensin II (AII) immunoreactivity in juxtaglomerular (JG) cells of rat kidney, which has been demonstrated previously by immunohistochemical studies, can be explained either as the product of intracellular synthesis or by the internalization of receptor-bound AII originating in plasma. To resolve these two alternative mechanisms, attempts were made to identify AI in JG cells of rat kidney by immunohistochemical staining using specific antibodies to AI. Although AI-like immunoreactivity was not detected in normal rat kidney, rats treated with the angiotensin-converting enzyme inhibitors, MK-421 or captopril, showed AI-like immunoreactivity in JG cells. The presence of renin and AII-like immunoreactivity was demonstrated in the same cells by specific antibodies to respective antigens used on adjacent serial sections. These findings support an intracellular mechanism of the formation of AII and suggest an intracellular renin angiotensin system, presumably separate from the extracellular system.
Asunto(s)
Angiotensina II/biosíntesis , Angiotensina I/biosíntesis , Angiotensinas/biosíntesis , Aparato Yuxtaglomerular/metabolismo , Animales , Captopril/farmacología , Dipéptidos/farmacología , Enalapril , Técnicas para Inmunoenzimas , Aparato Yuxtaglomerular/efectos de los fármacos , Masculino , Ratas , Ratas Endogámicas , Renina/biosíntesis , Sistema Renina-Angiotensina/efectos de los fármacosRESUMEN
Mouse hybridomas secreting antibodies to human insulin-like growth factor I (IGF I) were produced by fusion of spleen cells of hyperimmunised mice with FO mouse-myeloma cells. Eight clones producing antibodies against human IGF I have been isolated, two of which have been characterised. One was used in a radioimmunoassay, the other for immunopurification of IGF.
Asunto(s)
Anticuerpos Monoclonales , Insulina/sangre , Somatomedinas/sangre , Acromegalia/sangre , Animales , Complejo Antígeno-Anticuerpo , Línea Celular , Cromatografía Líquida de Alta Presión , Humanos , Insulina/deficiencia , Linfocitos/inmunología , Ratones , Ratones Endogámicos BALB C , Plasmacitoma , Radioinmunoensayo , Valores de Referencia , Somatomedinas/deficienciaRESUMEN
After the discovery of calretinin, a protein with high sequence homology to calbindin D-28k, the validity of immunohistochemical results obtained using polyclonal antibodies for this protein, was in question. In order to validate the previous results on the localization of calbindin D-28k in the brain, we localized the protein by highly specific monoclonal antibodies and revealed its mRNA histochemically by in situ hybridization. In general there was good agreement between the results obtained using these two different techniques and those reported in previous publications. The concordance was particularly impressive for the cerebral cortex, basal ganglia, basal nucleus of Meynert, hippocampus, thalamus, cerebellum and superior colliculus. In the amygdala and hypothalamus the low spatial resolution of in situ hybridization did not allow precise definition of some nuclei displaying a positive reaction for the protein. In the rhombencephalon, cells of the parabrachial nuclei and the dorsal raphe nucleus expressed calbindin D-28k. Neurons in the dorsal horn of the spinal cord and some horizontal cells of the retina were tagged with both methods. The only discrepancy was the presence of immunoreactive ependymal cells, whereas mRNA never occurred in cells lining the ventricles. Thus, the combined approach has established the widespread distribution of cells expressing calbindin D-28k in the rat brain.
RESUMEN
The serotoninergic input to the mammalian cerebral cortex originates in the median and the dorsal raphe nuclei. Median raphe neurons have been previously shown to give rise to beaded varicose axons which form dense pericellular arrays (baskets) surrounding the soma and the proximal dendrites of certain cortical neurons. In the present study, we have searched for specific markers characterizing the neurons of the marmoset neocortex and hippocampus surrounded by these thick varicose serotonin-containing fibers. The non-pyramidal nature of these neurons, suggested by their dendritic arborization, was correlated, in immunocytochemical experiments with double-labelling to demonstrate their surrounding serotonin-containing basket and their content of glutamic acid decarboxylase (GAD) or of the calcium-binding protein calbindin. Another calcium-binding protein common in numerous non-pyramidal cortical neurons, parvalbumin, was never found in neurons surrounded by serotonin-containing baskets. This organization was found in all areas of the neocortex and of the hippocampus where serotonin-containing baskets were present. One of the serotoninergic cortical inputs which originates from the brainstem tegmentum, traditionally described as "diffuse," proves to be highly selective in that a subset of its axons terminates preferentially on a subpopulation of inhibitory interneurons of the cerebral cortex. It may be emphasized that this subset of cortical interneurons has now been shown to be characterized not only by its axonal and dendritic arborization and its neurotransmitter, but also by a specific type of input which can modulate cortical function in a unique manner.
Asunto(s)
Callithrix/fisiología , Corteza Cerebral/fisiología , Hipocampo/fisiología , Interneuronas/fisiología , Proteína G de Unión al Calcio S100/metabolismo , Serotonina/metabolismo , Animales , Axones/metabolismo , Axones/fisiología , Calbindinas , Glutamato Descarboxilasa/metabolismo , Inmunohistoquímica , Parvalbúminas/metabolismoRESUMEN
The presence of neurones in the rat anterior medullary velum (AMV) has been investigated by using antibodies to the calcium-binding proteins, parvalbumin (PV), calretinin (CR), and calbindin-D28k (CB). Disparate populations of mainly GABAergic neurones were located in the rostral and caudal regions of the AMV. The rostral region of the AMV was characterised by GABAergic CR-labelled or PV-labelled neurones. CR-labelled neurones were bipolar or multipolar with round to ovoid somata (diameters between 8 and 12 microm), and rostrocaudally running dendrites forming a network. PV-labelled neurones had round somata (diameters between 6 and 10 microm) and were bi-tufted, with beaded dendrites. Both CR-labelled and PV-labelled dendrites formed punctate pericellular associations with unlabelled somatic profiles. In the caudal region of the AMV, PV-labelled neurones were GABAergic, multipolar cells, having round somata (diameters between 9 and 12 microm), with either beaded or nonbeaded dendrites forming a network of interconnecting dendrites. PV-labelled pericellular associations were made around both PV-labelled and unlabelled somatic profiles. CR labelled unipolar brush cells (UBCs) were not GABAergic. UBCs were characterised by a round to oval somata (10-15 microm in diameter) from which a single primary dendrite emerged to form a distal expansion having small terminal dendrites. From the distal expansion, there also appeared to be CR-labelled processes emanating and extending for up to 250 microm. CB occasionally labelled "Purkinje-like cells" (PLCs). The rat AMV is a more complex structure than first envisaged with the presence of predominantly inhibitory neurones expressing different calcium-binding proteins. Functional and anatomic aspects of this circuitry are further discussed.
Asunto(s)
Tronco Encefálico/química , Tronco Encefálico/citología , Cuarto Ventrículo/química , Cuarto Ventrículo/citología , Neuronas/química , Neuronas/citología , Animales , Calbindina 1 , Calbindina 2 , Calbindinas , Glutamato Descarboxilasa/análisis , Masculino , Parvalbúminas/análisis , Ratas , Ratas Wistar , Proteína G de Unión al Calcio S100/análisisRESUMEN
The macaque visual system has been frequently used as a model for understanding functional aspects of human vision. There are, however, few studies directly comparing biochemically defined neuronal populations in the visual cortex of the two species. In this study we compared the distribution and morphological features of the parvalbumin-immunoreactive neuronal subpopulation within humans and Old World monkeys (Macaca fascicularis and Macaca mulatta) by using monoclonal antibodies against the Ca2(+)-binding protein parvalbumin (PV), a neuronal marker in the vertebrate cerebral cortex. Characteristic laminar density and distribution of PV is observed, matching that seen with cytochrome C-oxidase and gamma-aminobutyric acid (GABA) immunoreactivity. Thus, parvalbumin is prominent in the layers receiving afferents from the dorsal lateral geniculate nucleus. Terminal fields are rich in layer IVA and IVC and moderate in the blob-region of layer II-III of the monkey cortex. In the human visual cortex only layer IVC displays rich terminal fields. Parvalbumin is present in neurons within all layers of the cortex except layer I. Parvalbumin-immunoreactive (PV-ir) axons occur in different lamellae of the white matter containing axons belonging to association or projection neurons. The estimation of PV-ir neurons, determined for 50 microns-wide columns through the thickness of area 17, shows that the percentage of the total neuron number in area 17 of humans containing PV is 6.8 +/- 2.0%, and in the macaque monkey, 11.5 +/- 2.9%. The perikaryal area of PV-ir neurons varies according to the layer and is comparable in humans (109.3 +/- 40.8 microns2) and monkeys (94.3 +/- 29.5 microns2). However, the relative number of large PV-ir neurons is higher in humans. The immunoreactive product fills the thinnest cell processes and the shape of PV-ir neurons can be easily traced with the aid of a camera lucida. The shape of the neurons is similar in the two species studied, and they probably belong to non-spiny stellate, double-bouquet, chandelier, and basket cell classes. This study shows that parvalbumin acts as a marker for a subpopulation of interneurons in area 17, but it is also present in the geniculocortical as well as in corticocortical pathways. Moreover, the Old World monkey and human visual cortices have a similar, but not identical, distribution of this important calcium-binding protein.