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The studies were carried out on the claustrum of 8 adult rabbits. Four types of neurons were distinguished: 1. Multipolar neurons, which have dendritic trunks either with conus (multipolar polygonal perikarya) or without conus (multipolar rounded perikarya). Both subdivisions of the multipolar neurons have 3–6 dendritic trunks. Only some branches of these trunks have spines. An axon emerges mainly from the cell body, rarely from the initial part of the dendritic trunk. 2. Bipolar neurons with fusiform or rounded perikarya; they have two dendrites covered with spines. An axon originates directly from the cell body or from one of the dendritic trunks. 3. Triangular neurons, which have three dendritic branches with spines. An axon emerges directly from the soma, often near the primary dendritic trunk. 4. Pear-shaped neurons with one or two dendritic trunks arise from one pole of the cell body and with an axon that originates from the opposite side of the perikaryon. The dendrites are covered with spines.
The connections between two parts of the claustrum in the rat and rabbit were studied using the highly fluorescent lipophilic carbocyanine dye (DiI). After the application of DiI crystal into the endopiriform nucleus, labeled fibers in the insular claustrum were observed in its part directly neighboring the insular cortex and capsula externa. Additionally, numerous projections into the piriform, insular and entorhinal cortices were present. The presence of connections between the endopiriform nucleus and insular claustrum suggests its role concerned with the processes taking part in the allocortical regions as well as in the limbic system.
Using axonal retrograde tracing, combined with morphometric analysis, we compared the distribution and number of claustral neurons projecting to the motor and somatosensory cortical areas in the Wistar rat. Comparable volumes of the retrograde tracer Fluoro-Gold, were injected into the motor or somatosensory cortices. Injections into these areas resulted in labeling of neurons along the entire length of the claustrum. Neurons retrogradely labeled after injection into the motor cortex prevailed in the anterior part of the claustrum, whereas those projecting to the somatosensory cortex predominated in the central part. The mean number of claustral neurons retrogradely labeled after tracer injections into the motor cortex significantly outnumbered that from the somatosensory cortical area (p<0.01). Similarly, the mean value of the numerical density of the retrogradely labeled neurons was significantly higher for the motor projection zone in the claustrum, than for the somatosensory projection zone (p<0.001). The contralateral claustral projections, both into the motor and somatosensory cortices, were considerably lower in number than the ipsilateral ones. These findings indicate that: (1) the claustral projections to the various cortical regions seem to be differentiated (2) the distribution of claustral neurons projecting to the motor and somatosensory neocortical areas shows an anteroposterior gradient, (3) the claustrum of the rat appears to be more closely related to the motor than to the somatosensory system, (4) the rat claustrum seems to function more as a satellite than a relay structure in relationship to the cerebral cortex.
An immunocytochemical double-staining method was applied in order to study the co-localisation of nitric oxide synthase (NOS) with three calcium-binding proteins, calbindin D28k (CB), calretinin (CR) and parvalbumin (PV) in the claustrum of the rat during the first 4 months of life (postnatal days: P0–P120). The co-localisation of NOS/PV and NOS/CB is reported. These neurons fall into the category of non-pyramidal cells. Double-labelled NOS/CB neurons are observed in the claustrum starting from P4, whereas double-labelled NOS/PV neurons are observed from P14 onwards. The percentages of double-labelled neurons increase in relation to the age. Double-labelled NOS/CB and NOS/PV neurons, although they do not constitute a numerous population, play an important role in the process of maturation of the claustrum. This is confirmed by the occurrence of these types of neurons at definite stages of maturation and by the increase in their number.
The quantitative analysis of the claustrocortical connections labeled with the fluorescent retrograde tracer Fluoro-Gold (FG) was conducted on 90 rabbits subdivided into the following age groups (P2, P7, P14, P21, P30, P60, P90, P120, P180). The equal volumes of retrograde fluorescent tracer FluoroGold (FG) were injected into the selected regions of the motor or somatosensory cortices. The volume of the dorsal part of the claustrum, total number of projecting neurons, numerical density and percentage distribution of projecting neurons were estimated by means of the unbiased stereological methods. The claustrocortical connections both with the motor and somatosensory areas in a rabbit are established in the postnatal life. The parts of the claustrum occupied by the motor and somatosensory projection zones as well as the morphology of the cortically projecting neurons do not reveal characteristic changes during the studied period. The significant decrease of the total number and numerical density of cortically projecting neurons as well as the increase of the claustral volume may reflect the process of adjustment of the claustrum to its modulatory function upon corresponding cortical areas. The intensity of the claustral connections with the motor and somatosensory cortices reveals significant difference during the studied period, being higher for the motor projection. It may be assumed that the claustrocortical connections established before birth undergo significant quantitative changes during postnatal development.
The morphological features of nitric oxide synthase (NOS)-containing neurons in the rat claustrum (Cl) were studied during the period of four months after birth. Forty-five animals divided into nine groups, according to survival period (P0, P4, P7, P10, P14, P21, P28, P60, P120) were used in the study. The immunocytochemical staining to neuronal NOS was performed and the material was studied both qualitatively and quantitatively using unbiased stereological methods. Our observations indicate that the process of maturation of NOS-immunoreactive (ir) neurons in Cl takes place during the early postnatal period. We report the increase of numerical density of immunoreactive neurons, changes in neuronal size, expressed by the decrease of the percentage of small neurons with simultaneous increase of the participation of medium-sized neurons and large neurons. In the whole studied period the prevalence of oval and fusiform neurons is observed. However, the increase of the proportion of multipolar neurons takes place. Round neurons are most characteristic in the youngest groups of animals and later become dominated by the developing subpopulations of irneurons of other shapes. In the anterior, central and posterior parts of Cl, a similar pattern of maturation of NOS-ir neurons is observed. No prevalence of characteristically shaped neurons is observed in any part of Cl. The adult-like pattern of morphological features in the NOS-ir neuronal population in Cl is reached in the third postnatal week. The maturation of NOS-ir neurons in the claustrum is a dynamic process which is not stabilised at the moment of birth. It may be assumed that characteristic changes of the NOS-ir population of neurons may be influential on the physiological processes observed in Cl. These may in particular have some importance for the processes of synaptogenesis and establishing as well as refining of numerous claustral connections with the other structures of the central nervous system.
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