RESUMEN
A simple and high-throughput transposon mediated mutagenesis system employing in vitro shuttle transposon mutagenesis has been used to systematically mutagenise the Streptomyces coelicolor genome. To achieve the highest coverage, a new ordered cosmid library was also constructed. Individual cosmids from both the existing and new libraries were disrupted using the Tn5-based mini-transposon Tn5062. A total of 35,358 insertions were sequenced resulting in the disruption of 6,482 genes (83% of the predicted open reading frames). Complete information for both the newly generated cosmids as well as all the insertions has been uploaded onto a central database, StrepDB ( http://strepdb.streptomyces.org.uk/ ). All insertions, new cosmids and a range of transposon exchange cassettes are available for study of individual gene function.
Asunto(s)
Elementos Transponibles de ADN/genética , Mutagénesis Insercional/métodos , Streptomyces coelicolor/genética , Cósmidos/genéticaRESUMEN
The purpose of these experiments was to determine the effects of cholinergic depletion on spontaneous and evoked activity of neurons in the different layers of the posteromedial barrel subfield (PMBSF) of the rat somatosensory cortex. Acetylcholine neurons in nucleus basalis of Meynert (NBM) were selectively lesioned with an immunotoxin (IT), 192 IgG-saporin. Spontaneous activity was significantly lower in layers II-III, Va, and VI in IT-injected animals compared to control animals. Evoked activity was significantly lower in layers II-III, IV, Vb, and VI of IT-injected animals compared to control animals. The largest difference was observed in layer Vb. Thus, cholinergic depletion causes significant changes in the magnitude of spontaneous and evoked activity but these differences are not completely in register with one another.
Asunto(s)
Acetilcolina/deficiencia , Núcleo Basal de Meynert/efectos de los fármacos , Colinérgicos/administración & dosificación , Neuronas/fisiología , Corteza Somatosensorial/fisiología , Potenciales de Acción/efectos de los fármacos , Potenciales de Acción/fisiología , Animales , Anticuerpos Monoclonales/administración & dosificación , Núcleo Basal de Meynert/citología , Potenciales Evocados/efectos de los fármacos , Inmunotoxinas/administración & dosificación , Inyecciones Intraventriculares , Masculino , Microinyecciones , N-Glicosil Hidrolasas , Neuronas/citología , Neuronas/efectos de los fármacos , Estimulación Física , Ratas , Ratas Sprague-Dawley , Proteínas Inactivadoras de Ribosomas Tipo 1 , Saporinas , Corteza Somatosensorial/citología , Corteza Somatosensorial/efectos de los fármacos , Vibrisas/inervación , Vibrisas/fisiologíaRESUMEN
Changes in market-driven health care economics are rapid and of great magnitude. This report describes a study of some of these changes in regard to children's health issues. We used a survey tool to assess long-range plans (next 10 years) and marketing strategies for major free-standing children's hospitals in different regions of the United States. We then used these assessments to draw conclusions about the impact of the plans and strategies on the practice of pediatric physicians and their workforce requirements. This may allow pediatric specialists and their programs to develop strategic plans and to take actions to contend with these market-driven economic changes. The tool was a questionnaire mailed to chief executive officers of 30 randomly chosen but geographically well-distributed children's hospitals. Seventeen children's hospitals responded (57%), providing information concerning each hospital and its current medical economic environment. The data were analyzed and trends were then identified from their responses. All institutions in this study expected to have fewer physicians on staff in the future. These institutions plan: (1) to improve quality and (2) to reduce costs. Quality will be improved by utilizing Continuous Quality Improvement (CQI) and/or Benchmarking to Best Practices, both of which encourage physicians to follow standardized treatment protocols. Costs will be reduced by decreasing hospital staff size. Some children's hospitals have merged or will merge with larger, full-service adult hospitals, but most plan to remain autonomous. Many expect a continued decrease in revenues, and almost all expect to downsize both bed number and staff. Restructuring will reduce the number of specialists, particularly in the fields of hematology-oncology, psychiatry, endocrinology, nephrology, and cardiology, and will also reduce the number of surgical specialists. The administrators predicted that more nurse practitioners will be employed at these hospitals in the 21st century, serving the role of physician extenders, if not physician substitutes. To ensure their own survival, physicians must develop their own strategic plans as have the hospitals in which they practice. Optimally they should embrace those activities that parallel their hospital's efforts such as Quality Improvement (CQI, Benchmarking), cost containment, and practice mergers.
Asunto(s)
Hospitales Pediátricos , Pediatría/tendencias , Adulto , Niño , Predicción , HumanosRESUMEN
To improve transposon mutagenesis of antibiotic-producing Streptomyces, a mini-transposon, Tn1792, was constructed, based on IS6100, originally isolated from Mycobacterium fortuitum. Easily manageable transposition assays were developed to demonstrate inducible transposition of Tn1792 into the Streptomyces genome from a temperature-sensitive delivery plasmid. Introduction of the selectable aac1 gene between the inverted repeats in Tn1792 allowed for both reliable identification of transposition events in Streptomyces, and also subsequent cloning of transposon-tagged sequences in Escherichia coli. This enabled the target site specificity of Tn1792 to be determined at nucleotide resolution, revealing no significant shared homology between different target sites. Consequently, Tn1792 is well suited for random mutagenesis of Streptomyces.
Asunto(s)
Elementos Transponibles de ADN/genética , Genes Bacterianos/genética , Mutagénesis Insercional/métodos , Streptomyces/genética , Antibacterianos/biosíntesis , Secuencia de Bases , Clonación Molecular , Regulación Bacteriana de la Expresión Génica/genética , Genoma Bacteriano , Datos de Secuencia Molecular , Plásmidos , Regiones Promotoras Genéticas , Streptomyces/metabolismoRESUMEN
Degeneration of cholinergic neurons in the basal forebrain is a neural marker of Alzheimer's disease and is associated with perceptual and cognitive deficits. An idea that has attracted scientific scutiny is that aging makes the brain more susceptible to neurodegenerative diseases such as Alzheimer's. The purpose of this study was to compare the effects of the loss of cholinergic input from nucleus basalis of Meynert on evoked activity in the posteromedial barrel subfield of the somatosensory cortex in young (2-2.5 months) and aged (28-30 months) male Fisher hybrid rats. The mean firing rate and receptive fields of single neurons in the posteromedial barrel subfield of the somatosensory cortex were examined after selective lesions of cholinergic neurons in the nucleus basalis of Meynert with an immunotoxin. IgG 192-saporin. Functional properties of single neurons in young animals were affected much more significantly by cholinergic depletion than those in aged animals. In cholinergic-depleted young animals, the mean firing rate of evoked activity and receptive field of posteromedial barrel subfield neurons were significantly decreased. Cholinergic depletion caused a 14% decrease in evoked activity and a 33% increase in receptive field size in young animals. The mean firing rate and receptive field of single neurons were not affected by cholinergic depletion in aged animals. It is concluded that functional properties of cortical sensory neurons in young animals are more vulnerable to cholinergic depletion than are those of aged animals and that cholinergic depletion does not further impact the properties of neurons exposed to the processes of aging.
Asunto(s)
Acetilcolina/deficiencia , Envejecimiento/fisiología , Potenciales Evocados Somatosensoriales/fisiología , Neuronas/fisiología , Sustancia Innominada/fisiología , Animales , Anticuerpos Monoclonales , Inmunotoxinas , Inyecciones Intraventriculares , Masculino , N-Glicosil Hidrolasas , Vías Nerviosas/fisiología , Ratas , Ratas Endogámicas F344 , Proteínas Inactivadoras de Ribosomas Tipo 1 , Saporinas , Sustancia Innominada/citologíaRESUMEN
The aim of this study was to assess the distribution of neurons immunoreactive for parvalbumin (PV), calbindin (CaBP), glutamic acid decarboxylase (GAD), and gamma-aminobutyric acid (GABA) in the somatosensory thalamus of the raccoon and to compare these features to those of other species, especially primates. Immunohistochemistry was used to study the location of these neurons in the ventroposterior nucleus (VP), ventroposterior inferior nucleus (VPI), posterior group of nuclei (Po), and reticular nucleus (Rt). A consistent differential pattern of PV-positive (PV+) and CaBP-positive (CaBP+) cells was found in the somatosensory thalamus. Many PV+ neurons were observed in VP and Rt, but very few were found in VPI or Po. In contrast, CaBP+ neurons were distributed throughout VP, VPI, and Po but were very sparse or absent in Rt. In the VP nucleus, PV+ cells were distributed in clusters separated by interclusteral regions with a sparse distribution of PV+ cell bodies. The distributions of PV+ and CaBP+ cells tended to be complementary to each other in VP; regions with a high density of PV+ neurons had a low density of CaBP+ cell bodies. Double-labeling experiments revealed very few neurons in which PV and CaBP immunoreactivities were colocalized. Cells immunoreactive for GAD or GABA were found in PV-dense clusters of VP; fewer GABAergic neurons were present in the CaBP-dense interclusteral regions of VP and in VPI and Po. GAD+ and GABA+ neurons were most prominently distributed in Rt. We conclude that the distributions of PV+ and CaBP+ cell bodies in the raccoon somatosensory thalamus are very similar to those in primates. The density of GABAergic neurons in the somatosensory thalamus of the raccoon is less than that in the cat and monkey, but the relative distribution of GABAergic neurons in the different somatosensory nuclei is very similar to that in the cat and monkey. These results are discussed in relation to findings in other species and are related to the functions of lemniscal and nonlemniscal somatosensory pathways.
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Neuronas/citología , Parvalbúminas/análisis , Mapaches/anatomía & histología , Proteína G de Unión al Calcio S100/análisis , Núcleos Talámicos/citología , Tálamo/citología , Animales , Calbindinas , Gatos , Glutamato Descarboxilasa/análisis , Haplorrinos , Inmunohistoquímica , Proteínas del Tejido Nervioso/análisis , Primates , Especificidad de la Especie , Núcleos Talámicos/anatomía & histología , Tálamo/anatomía & histología , Ácido gamma-Aminobutírico/análisisRESUMEN
Clinical and functional studies have strongly suggested that acetylcholine input from the nucleus basalis of Meynert is important for the cortex's adaptive response to experience. The purpose of this study was to investigate the effects of depletion of acetylcholine inputs from nucleus basalis of Meynert on experience-dependent plasticity in the cortex of young adult male rats. The posteromedial barrel subfield in the primary somatosensory cortex was studied. Experience-dependent plasticity was elicited using a whisker-pairing paradigm in which all whiskers except D2 and D3 were trimmed daily. Plasticity within barrel D2 of the posteromedial barrel subfield was measured using the electrophysiological extracellular recording technique. An index of plasticity was determined in two ways: as an increase in the magnitude of evoked activity to stimulation of whisker D2 and as a bias in the ratio of evoked activity for stimulation of paired whisker D3 and cut whisker D1 (D3/D1). Whiskers D2, D3 and D1 were stimulated (deflected) by a Chubbuck electromechanical stimulator. Cholinergic neurons in the nucleus basalis of Meynert were selectively lesioned with an immunotoxin, 192 IgG-saporin, injected into the left lateral ventricle. Lesions of cholinergic neurons in the nucleus basalis of Meynert were verified using choline acetyltransferase immunocytochemistry and radioenzymatic assay. Experience-dependent plasticity was significantly reduced in cholinergic-depleted animals. The magnitude of evoked activity to stimulation of whisker D2 increased by 16-100% in control animals compared with 0-20% in cholinergic-depleted animals. Similarly, compared to a 60-100% increase in the D3/D1 ratio of evoked activity for phosphate-buffered saline-injected control animals, cholinergic-depleted rats showed no significant increase in the D3/D1 ratio (0-15%) after undergoing the whisker-pairing paradigm. After whisker trimming, the D3/D1 response ratio in immunotoxin-treated animals was essentially the same as in control animals that had not been subjected to the whisker-pairing paradigm. This study showed that no significant plasticity response was observed in the absence of cholinergic input from the nucleus basalis of Meynert. The mechanisms of the action of acetylcholine in cortical plasticity are still not known, but we hypothesize that this type of plasticity is activity dependent and is significantly enhanced in the presence of acetylcholine.
Asunto(s)
Acetilcolina/fisiología , Anticuerpos Monoclonales/farmacología , Inmunotoxinas/farmacología , Plasticidad Neuronal/fisiología , Neuronas/fisiología , Corteza Somatosensorial/fisiología , Sustancia Innominada/fisiología , Vibrisas/fisiología , Acetilcolina/análisis , Animales , Anticuerpos Monoclonales/administración & dosificación , Mapeo Encefálico , Ventrículos Cerebrales/efectos de los fármacos , Ventrículos Cerebrales/fisiología , Colinérgicos/administración & dosificación , Colinérgicos/farmacología , Electrofisiología/métodos , Potenciales Evocados , Inmunotoxinas/administración & dosificación , Inyecciones Intraventriculares , Masculino , N-Glicosil Hidrolasas , Plasticidad Neuronal/efectos de los fármacos , Neuronas/efectos de los fármacos , Estimulación Física , Ratas , Ratas Sprague-Dawley , Proteínas Inactivadoras de Ribosomas Tipo 1 , Saporinas , Corteza Somatosensorial/efectos de los fármacos , Sustancia Innominada/efectos de los fármacos , Vibrisas/inervaciónRESUMEN
The purpose of these experiments was to determine the effects of cholinergic depletion on the morphology and staining density of barrels formed by glutamic acid decarboxylase-positive neuropil in the posteromedial barrel subfield of the somatosensory cortex. The density and distribution of glutamic acid decarboxylase immunoreactive neuropil were examined after highly selective lesions of cholinergic neurons in the nucleus basalis of Meynert with an immunotoxin, IgG 192-saporin. Glutamic acid decarboxylase immunoreactivity was also examined in animals subjected to a whisker-pairing experience and lesion of acetylcholine inputs from the nucleus basalis of Meynert. Seven to 9 weeks after intraventricular injection of the immunotoxin, animals were perfused with a zinc aldehyde fixative and glutamic acid decarboxylase immunoreactivity was examined in 30-micron tangential sections. Cholinergic depletion caused reduced glutamic acid decarboxylase immunoreactivity in selective regions of the posteromedial barrel subfield. The density of neuropil and cell bodies immunoreactive for glutamic acid decarboxylase was significantly reduced in septa and perimeters of barrel walls. The length, width, and area of barrels were reduced 10-20% in cholinergic-depleted animals compared with controls. The density of glutamic acid decarboxylase immunoreactivity in the hollow of barrels was not affected by this treatment. Whisker pairing did not significantly change the density of glutamic acid decarboxylase immunoreactivity in barrels. These observations are discussed in regard to how long-term cholinergic depletion affects the function of different fiber systems in the posteromedial barrel subfield cortex and how some sensory functions may be comprised.
Asunto(s)
Acetilcolina/fisiología , Glutamato Descarboxilasa/metabolismo , Corteza Somatosensorial/enzimología , Acetilcolina/deficiencia , Animales , Inmunohistoquímica , Inmunotoxinas/farmacología , Masculino , Ratas , Ratas Sprague-Dawley , Vibrisas/fisiologíaRESUMEN
The ventral forebrain, including the ventral striatum, the ventral pallidum and the substantia innominata, is an important region involved in the functions of the basal ganglia and the limbic system, as well as the magnocellular corticopetal neurons of the nucleus basalis of Meynert. Although previous studies have shown that this region is richly innervated by GABAergic fibers, little is known with respect to the relative densities of GABAergic to non-GABAergic axon terminals in this region. To address this issue, we have developed a specific rabbit antiserum to GABA and used a postembedding immunocytochemical reaction to analyse the distribution of GABA-like immunoreactive axon terminals in the rat ventral striatum, ventral pallidum and substantia innominata. Of all axon terminals that form identifiable synapses within single ultrathin sections taken from these regions, 11.6% in the ventral striatum, 85.5% in the ventral pallidum and 64.8% in the substantia innominata were GABAergic. Differences were also found in the distribution patterns of these terminals with respect to the size of their synaptic target dendrites. These findings are consistent with previous findings that a majority of inputs to the ventral striatum are excitatory, and that a majority of inputs to the ventral pallidum are inhibitory. Our results provide a first approximation of the anatomical substrate for the physiology and pharmacology of GABA actions in the ventral forebrain region. These results also show that GABA may play an important role in the substantia innominata, where both the cholinergic and the non-cholinergic magnocellular corticopetal neurons reside within a neuropil innervated by many different non-cholinergic fibers.
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Axones/fisiología , Axones/ultraestructura , Prosencéfalo/fisiología , Prosencéfalo/ultraestructura , Ácido gamma-Aminobutírico/fisiología , Animales , Especificidad de Anticuerpos , Hemocianinas/inmunología , Inmunoconjugados/inmunología , Inmunohistoquímica , Microscopía Electrónica , Neostriado/fisiología , Neostriado/ultraestructura , Ratas , Ratas Sprague-Dawley , Sustancia Innominada/fisiología , Sustancia Innominada/ultraestructuraRESUMEN
We investigated (1) the topography of projection neurons in the nucleus basalis of Meynert (NBM) with efferents to restricted regions of the primary somatosensory (SI), the second somatosensory (SII), and the primary motor (MI) cortices in the rat; (2) the percentage of these NBM projection neurons that were cholinergic; and (3) the collateralization, if any, of single NBM neurons to different subdivisions within SI, to homotopic areas of SI and SII, and to homotopic areas of SI and MI. Retrograde single- and double-labeling techniques were used to study NBM projections to electrophysiologically identified subdivisions of SI and to homotopic representational areas of SI and SII, and of SI and MI. Choline acetyltransferase immunocytochemistry was done to identify cholinergic NBM neurons. Of the retrogradely labeled NBM neurons that projected to selective subdivisions of SI, SII, and MI, 89%, 87%, and 88%, respectively, were cholinergic. We found a rostral-to-caudal progression of retrogradely labeled NBM neurons following a medial-to-lateral sequence of injections into subdivisions of SI. Overlapping groups of single-labeled NBM neurons were observed after injections of different tracers into adjacent subdivisions within SI or homotopic areas of SI and SII, and of SI and MI. We conclude that NBM innervation to SI, SII, and MI is mostly cholinergic in the rat, that each cortical area receives cholinergic afferents from neurons widely distributed within the NBM, and that each NBM neuron projects to a restricted cortical area without significant collateralization to adjacent subdivisions within SI or to homotopic areas of SI and SII, or SI and MI.
Asunto(s)
Acetilcolina/fisiología , Mapeo Encefálico , Neuronas Aferentes/fisiología , Ratas Sprague-Dawley/fisiología , Corteza Somatosensorial/fisiología , Sustancia Innominada/fisiología , Animales , Toxina del Cólera , Colorantes Fluorescentes , Miembro Anterior/inervación , Masculino , Neuronas Aferentes/química , RatasRESUMEN
The growth and activity of Streptomyces violaceolatus and Streptomyces lividans was studied in soil under controlled conditions. The life cycle was followed under differing nutrient regimes and the fate of plasmid- and phage-borne genes determined by direct and indirect techniques. Methods were developed for the direct monitoring of plasmid DNA extracted from soil which allowed differentiation of the cellular location of plasmid DNA between mycelium and spores. In a dynamic, nutrient-fed soil microcosm, inoculants survived poorly, but a specific stage was defined by direct and indirect methods when the inoculants were most active and this correlated with the detection of gene transfer events. Plasmid transfer, phage infection and lysogeny only occurred to a significant extent within this stage at days 15-17 during a 60-day incubation. Estimates based on plasmid DNA recovery indicated that viable counts underestimated spore and mycelial propagules by a factor of greater than 100.
Asunto(s)
Microbiología del Suelo , Streptomyces/genética , Transfección , Bacteriófagos/fisiología , ADN Bacteriano/aislamiento & purificación , Monitoreo del Ambiente , Plásmidos , Esporas Bacterianas/genética , Streptomyces/crecimiento & desarrollo , Streptomyces/fisiologíaRESUMEN
1. We examined whether the three physiologically defined neuron types described for rodent neocortex were also evident in human association cortex studied in an in vitro brain slice preparation. We also examined the relationship between physiological and morphological cell type in human neocortical neurons. In particular, we tested whether burst-firing neurons were numerous in regions of human cortex that are susceptible to seizures. 2. Although we sampled regular-spiking and fast-spiking neurons, we observed no true burst-firing neurons, as defined for rodent cortex. We did find neurons that displayed a voltage-dependent shift in firing behavior. Because this behavior was due, in large part, to a low-threshold calcium conductance, we called these cells low-threshold spike (LTS) neurons. 3. Regular-spiking neurons and LTS neurons only differed in the voltage dependence of firing behavior and the first few interspike intervals (ISIs) of repetitive firing in response to small current injections (from hyperpolarized membrane potentials). Because of the general similarities between the two types, we consider the LTS cells to be a subgroup of regular-spiking cells. 4. All biocytin-filled regular-spiking neurons were spiny and pyramidal and found in layers II-VI. The lone filled fast-spiking cell was aspiny and nonpyramidal (layer V). The LTS neurons were morphologically heterogeneous. We found 80% of LTS neurons to be spiny and pyramidal, but 20% were aspiny nonpyramidal cells. LTS neurons were located in layers II-VI. 5. In conclusion, human association cortex contains two of three physiological cell types described in rodent cortex: regular spiking and fast spiking. These physiological types corresponded to spiny, pyramidal, and aspiny, nonpyramidal cells, respectively. We sampled no intrinsic burst-firing neurons in human association cortex. LTS neurons exhibited voltage-dependent changes in firing behavior and were morphologically heterogeneous: most LTS cells were spiny and pyramidal, but two cells were found to be aspiny and nonpyramidal. It is not clear whether the absence of burst-firing neurons or the morphological heterogeneity of LTS neurons are due to species differences or differences in cortical areas.
Asunto(s)
Aprendizaje por Asociación/fisiología , Corteza Cerebral/fisiología , Neuronas/fisiología , Axones/ultraestructura , Corteza Cerebral/fisiopatología , Estimulación Eléctrica , Electrofisiología/métodos , Epilepsia/fisiopatología , Epilepsia/cirugía , Potenciales Evocados , Humanos , Técnicas In Vitro , Potenciales de la Membrana , Neuronas/ultraestructura , Tractos Piramidales/citología , Tractos Piramidales/fisiología , Factores de TiempoRESUMEN
A new method for the isolation and enumeration of streptomycete spores from soil was developed. This method makes use of a cation-exchange resin to disperse soil particles. It allowed the detection of 10 spores in 100 g of sterile soil, while ca. 10 could be accurately enumerated in 100 g. This method was applied to studying the fate of a marked actinophage in soil. In sterile amended and nonsterile soil, relatively high numbers of actinophages were only found during the first few days of the experiment when the host streptomycete was in the mycelial form. Later, after sporulation, lysogens could be detected in sterile amended soil and could still be found 60 days after inoculation. Although no lysogens were found in nonsterile soil, the introduced phage could still be detected in the free state after 60 days, albeit at a low titer.
RESUMEN
The organization of intracortical and callosal projecting cell bodies was examined in somatosensory representation areas I (SI) and II (SII) of the raccoon by use of horseradish peroxidase (HRP) or horseradish peroxidase-wheat germ agglutin (HRP-WHA). HRP and HRP-WHA were injected into commissurally and noncommissurally connected subdivisions of SI and SII. Injection sites in SII were identified electrophysiologically. Results were obtained from transverse sections in which the HRP was visualized with the aid of the substrates dihydrochlorobenzidine or tetramethyl benzidine in the presence of hydrogen peroxidase. The principal findings were the following: (1) there are reciprocal connections between SI and SII; (2) in SI the intracortically projecting cell bodies and terminals are located primarily in sulcal cortex; (3) intracortically projecting neurons in SI are located primarily in layers III whereas in SII they are located principally in layers III and V; (4) there are connections between disparate areas within SI; and (5) there are intracortical connections between callosum-connected and acallosal regions in SII. These results are discussed with regard to the results of mapping studies of the SI, the significance of intracortical connections to the formation of sulci in SI, and the possible roles of nonhomotopic connections in the intermanual transfer of learning.
Asunto(s)
Cuerpo Calloso/anatomía & histología , Mapaches/anatomía & histología , Corteza Somatosensorial/anatomía & histología , Animales , Mapeo Encefálico , Peroxidasa de Rábano Silvestre , Vías Nerviosas/anatomía & histología , Aglutininas del Germen de TrigoRESUMEN
The submodality and receptive field properties of single units in the lateral cervical nucleus (LCN) of barbiturate anesthetized cats were studied with glass microelectrodes. In other experiments, a region of the posterior thalamus containing neurons with properties comparable to those seen in the LCN was examined with tungsten microelectrodes. The responses of most units in the LCN reflected a major input from large myelinated afferent fibers innervating guard hairs but no input from Pacinian afferent fibers. The large size of the receptive fields indicated that excitatory input converged selectively from afferent fibers serving hairs over large areas of the body. In the posterior thalamus rapidly adapting neurons characterized by very large receptive fields and driven by the movement of guard hairs were observed to a region identified histologically as the rostral extension of the lateral division of the posterior nuclear group (POl). Caudally this region was located immediately adjacent to the dorsolateral part of the ventroposterior inferior nucleus (VPI). In more rostral parts of the thalamus it was located more dorsally and the ventroposterior lateral nucleus intervened between it and the VPI. This region was less than 1 mm wide in the frontal plane but extended rostrocaudally for several millimeters. Horseradish peroxidase injected into the region of the VPI and the POl labeled many cells in the LCN and the caudal pole of the dorsal column nuclei demonstrating that neurons in the LCN relay information to this part of the thalamus. These data, plus previous experiments showing that the VPI receives a major projection from the caudal poles of the dorsal column nuclei, suggest that the rostral portion of the POl receives an important afferent supply from the LCN. The responses of neurons in the POl appear to arise from specific classes of sensory receptors and cannot be considered less precise or more primitive than responses observed in the ventroposterior nucleus of the thalamus.
Asunto(s)
Mecanorreceptores/fisiología , Médula Espinal/fisiología , Núcleos Talámicos/fisiología , Vías Aferentes/fisiología , Animales , Mapeo Encefálico , Gatos , Femenino , Articulaciones/inervación , Masculino , Músculos/inervación , Neuronas/fisiología , Nociceptores/fisiología , Corpúsculos de Pacini/fisiología , Piel/inervación , Corteza Somatosensorial/fisiología , Raíces Nerviosas Espinales/fisiología , Transmisión SinápticaRESUMEN
The adequate stimulus and body site that excited neurons in cat cortical somatosensory areas 3a and 3b were recorded using low-impedance tungsten microelectrodes. Horizontal penetrations provided a good correlation between the electrophysiological and cytoarchitectonic data. Responses best driven by cutaneous stimuli were replaced with responses driven by manipulation of deep tissue at, or very near, the border between areas 3a and 3b. Following functional identification of these areas horseradish peroxidase was injected into one of them. Injections into area 3a labeled neurons in a rostral and dorsal cap of the ventroposterior thalamus. It was suggested that this region is a distinct nucleus termed the ventroposterior oralis nucleus (VPO). Injections into the forelimb portion of area 3b labeled neurons in the ventroposterior lateral nucleus (VPL). With both vertical and horizontal microelectrode trajectories through the ventroposterior thalamic nuclei, inputs from deep structures presumed to be muscles were consistently located in the VPO nucleus and cutaneous inputs activated neurons in the VPL. The existence of several functionally-distinct subdivisions within the somatosensory nuclei of the thalamus supports the hypothesis of parallel processing and relay of somatosensory information at this level of the pathway.
Asunto(s)
Corteza Somatosensorial/fisiología , Núcleos Talámicos/fisiología , Vías Aferentes/fisiología , Animales , Mapeo Encefálico , Gatos , Potenciales Evocados Somatosensoriales , Femenino , Articulaciones/inervación , Masculino , Mecanorreceptores/fisiología , Músculos/inervación , Neuronas/fisiología , Corpúsculos de Pacini/fisiología , Piel/inervación , Transmisión SinápticaRESUMEN
The ventroposterior region of the thalamus of mongrel cats was searched to locate zones activated by somatic stimuli. By using stimuli that selectively excited Pacinian corpuscles, areas activated by this class of afferent fibers were differentiated from regions activated by other classes of cutaneous mechanoreceptors. The results showed that Pacinian inputs excite neurons in the ventroposterior inferior nucleus (VPI) of the thalamus, whereas other more dorsal zones within the ventroposterior thalamus receive inputs from other mechanoreceptor classes. This definition of the VPI tended to be larger and to extend further lateral than some published descriptions. Horseradish peroxidase (HRP) was injected into ventroposterior zones shown by electrophysiological recordings to receive inputs from Pacinian afferents. Subsequently, labeled cell bodies were observed in the caudal poles of the dorsal column nuclei, a region previously shown to be activated by Pacinian afferents. Very few labeled cells were found in the central region of these nuclei, a region previously shown to be activated by other classes of cutaneous mechanoreceptors. Electrophysiological recordings were used to locate a small portion of the second somatosensory cortex driven by Pacinian stimuli. When HRP was injected into this region cell bodies in the VPI and the lateral part of the posterior group were labeled, but few or no labeled cells were found in ventroposterior lateral nucleus. We hypothesize that the VPI receives Pacinian information from a cytoarchitecturally distinct region in the caudal poles of the dorsal column nuclei. Further, we suggest that a major cortical target for the VPI is a subdivision of the second somatosensory cortex. These studies do not exclude the possibility that Pacinian inputs have other thalamic and cortical targets.
Asunto(s)
Mecanorreceptores/fisiología , Corpúsculos de Pacini/fisiología , Corteza Somatosensorial/fisiología , Núcleos Talámicos/fisiología , Vías Aferentes/anatomía & histología , Vías Aferentes/fisiología , Animales , Mapeo Encefálico , Gatos , Femenino , Masculino , Neuronas/fisiología , Corpúsculos de Pacini/anatomía & histología , Corteza Somatosensorial/anatomía & histología , Núcleos Talámicos/anatomía & histologíaRESUMEN
The thalamocortical afferents to cortical somatosensory areas I (SI) and II (SII) were investigated in the raccoon using the horseradish peroxidase technique. The purpose of this study was to determine if the cell bodies of origin for thalamocortical afferents to these cortical regions were localized in the same or different nuclei in the ventroposterior region of the thalamus. Horseradish peroxidase was injected into subdivisions of SI or SII and after post-injection survival periods of 12-72 hours the horseradish peroxidase in the tissue was reacted with the chromogens dihydrochlorobenzidine or tetramethylbenzidine in the presence of hydrogen peroxide. The results show that SI and SII receive projections from neurons in separate and distinct nuclei in the ventroposterior thalamus. Following injections into subdivisions of area I, a topographical distribution of retrogradely-labelled cell bodies was observed in the ventrobasal complex. Following injections of horseradish peroxidase into subdivisions of area II, a topographical distribution of labelled cell bodies was observed in the ventroposterior inferior nucleus. No labelled cell bodies were observed in the ventrobasal complex. The thalamocortical connections of somatosensory cortices I and II in raccoon are compared with those in other animals and it is suggested that these two cortical areas may be involved in differential processing of tactile information.
Asunto(s)
Mapaches/anatomía & histología , Corteza Somatosensorial/anatomía & histología , Núcleos Talámicos/anatomía & histología , Vías Aferentes/anatomía & histología , Animales , Miembro Anterior/inervación , Miembro Posterior/inervación , Peroxidasa de Rábano SilvestreRESUMEN
A rapid, inexpensive method is described for achieving accurate reproduction of histological sections. The method uses a microfiche reader-printer (for library use) which produces A4 size prints. Interchangeable lenses on the microfiche printer permit the magnification of the section to be varied over the range 6.6-72 times. Where large numbers of sections are involved the speed and low cost of the method offers considerable advantages over traditional hand tracing or photomicrography.