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Intracerebral haemorrhage is a devastating neurological disease with high mortality rate and poor prognosis. The most prominent manifestation of the disease are the movement disorders, but many patients also suffer from cognitive impairment. Taking into account vulnerability of the neurons located within the hilus of the dentate gyrus (HDG) to many brain insults we decided to study the effect of experimentally induced intracerebral haematoma on density of neurons expressing NogoA protein in HDG. In addition, we studied how administration of valproic acid and minocycline, the two drugs generally believed to be neuroprotective agents, influences the density of these neurons. Our study revealed that 4 weeks after intracerebral haematoma induction, minocycline and valproic acid treatment increased the densities of NogoA-ir neurons in the hilus of contralateral dentate gyrus once the data were compared to ipsilateral hemispheres within the same group. The analysis of contralateral hemisphere data, however, revealed increased densities of NogoA-positive neurons in haematoma and valproic acid treated animals when compared to contralateral hemispheres of control animals. The administration of minocycline was, however, able to alleviate this increase. These changes may influence the haematoma-induced reorganisation of neuronal circuitries in the dentate gyrus. (Folia Morphol 2014; 73, 3: 279–285)
Intracerebral hemorrhage (IH) is a devastating form of cerebrovascular disease. Several studies points at glial response as one of the key factors in processes related to brain damage and repair after IH. The aim of the present study was to evaluate the influence of administration of minocycline and valproic acid on glial response after IH. 56 adult male Wistar rats (290 - 410 g) were used for the study. In 48 rats IH was induced by injection of 200 µl of autologous blood into the temporal lobe structures. Next animals were divided into three groups that received either minocycline (3 × 200 mg/kg), or valproic acid (2 × 45 mg/kg) or 0.09% saline (2 ml) for 7 consecutive days. 8 sham-operated animals served as controls. Animals from each group were successively sacrificed at 2, 4, 24 and 48 weeks after the hematoma induction. The intensity of glial response was based on qualitative and semiquantitative evaluations of immunostained sections. We observed that during the course of IH glial cells exhibit time-dependent changes in morphology and intensity of staining. 2 and 4 weeks after IH induction activated forms of astro- and microglia were observed near the border of hematoma as well as neighboring structures, while 24 and 48 weeks later they were present mainly around the glial scar and in the degenerating white matter. White matter structures also contained NOGO-A-immunoreactive oligodendrocytes. Administration of minocycline or valproic acid decreased the number of activated astro- and microglial cells in the white matter of hemisphere contralateral to site of injury at all time points. The number of oligodendrocytes was influenced only by minocycline. The obtained results indicate that minocycline and valproic acid administration modifies the number of glial cells in the white matter of hemisphere contralateral to the site injury. The study was financially supported by Polish Ministry of Science and Higher Education grant nr 3419/B/P01/2008/35.
BACKGROUND AND AIMS: The study evaluates the effect of experimentally induced intracerebral haematoma (ICH) on density of neurodegenerating neurons and volume of the rat striatum. In addition, we studied how administration of valproic acid and minocycline, the two drugs generally believed to be neuroprotective agents, influences the density of these neurons. METHODS: 80 adult male Wistar rats were assign to one of 4 experimental groups: control, (CON), haematoma (HAEM), haematoma and minocycline (MINO) and haematoma and valproic acid (VAL). Five animals from each experimental group were sacrificed at four time points: 2 weeks, 4 weeks, 24 weeks and 48 weeks after ICH. To assess the neurodegeneration the sections were stained with FluoroJadeB (FJB). The CellSense Dimension 1.5 (Olympus, Japan) image analysis system was employed to measure the densities of FJB-labelled neurons as well as volume of the striatum. Statistical analyses were performed using two way ANOVA with replications. To find differences between groups post-hoc Tukey test was applied. A P-value <0.05 was considered significant. RESULTS: Our study revealed that valproic acid treatment decreased the density of FJB-labeled degenerating neurons 4 weeks after ICH induction in the ipsilateral striatum. Minocycline did not have such effect. None of the two drugs influenced the volume of the striatum. CONCLUSIONS: Our study revealed that valproic acid alleviates striatal neuron degeneration in the rat model of ICH. Because striatum has been implicated to play role in motor control as well as cognition and affective control, the valproic acid treatment might have beneficial effects on these processes after ICH.
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.
This study describes the topography, borders and divisions of the globus pallidus in the Brazilian short-tailed opossum (Monodelphis domestica) and distribution of the three calcium binding proteins, parvalbumin (PV), calbindin D-28k (CB) and calretinin (CR) in that nucleus. The globus pallidus of the opossum consists of medial and lateral parts that are visible with Nissl or Timm’s staining and also in PV and CR immunostained sections. Neurons of the globus pallidus expressing these proteins were classified into three types on the basis of size and shape of their soma and dendritic tree. Type 1 neurons had medium-sized fusiform soma with dendrites sprouting from the opposite poles. Neurons of the type 2 had medium-to-large, multipolar soma with scarce, thin dendrites. Cell bodies of type 3 neurons were small and either ovoid or round. Immunostaining showed that the most numerous were neurons expressing PV that belonged to all three types. Density of the PV-immunopositive fibers and puncta correlated with the density of the PV-labeled neurons. Labeling for CB resulted mainly in the light staining of neuropil in both parts of the nucleus, while the CB-expressing cells (mainly of the type 2) were scarce and placed only along the border of the globus pallidus and putamen. Staining for calretinin resulted in labeling almost exclusively the immunoreactive puncta and fibers that were distributed with medium-to-high density throughout the nucleus. Close to the border of globus pallidus with the putamen these fibers (probably dendrites) were long, thin and varicous, while more medially bundles of thick, short and smooth fibers predominated. Single CR-ir neurons (all of the type 3) were scattered through the globus pallidus. Colocalization of two calcium binding proteins in one neuron was never observed. The CB-ir puncta (probably terminals of axons projecting to the nucleus) frequently formed basket-like structures around the PV-ir neurons. Therefore, the globus pallidus in the opossum, much as that in the rat, consists of a heterogeneous population of neurons, probably playing diversified functions.
To understand the organization of inhibitory circuitries in the striatum of the opossum, the distribution of parvalbumin (PV), calretinin (CR) and calbindin (CB) was investigated within the nucleus accumbens (Acc), caudate nucleus (Cd) and putamen (Pu). Brains from six adult opossum (both sexes) were stained for PV, CR and CB and analyzed using fl uorescent or confocal microscopy. Within neurons immunostained for each calcium-binding protein four major types of neurons were distinguished. Type 1 – small ovoid or roundish somata with three to fi ve thin dendrites of approximately equal thickness; type 2 – medium-to-large-sized multipolar neurons with dendrites of variable thickness; type 3 – fusiform neurons of variable size emanating dendrites from the opposite poles of the somata; type 4 – pyramidal neurons. Moderate-to-high density of CB immunoreactive (-ir) neurons was observed in the Cd and Pu, but staining in the Acc was lighter. Light density of PV-ir neurons was present in the Cd, Pu and Acc, but PV-immunoreactivity of neuropil was high. Only single CR-ir neurons were scattered through the all studied structures, but the immunostaning of neuropil was much higher. Our data provide baseline information for comparisons of distribution of calcium binding proteins in different species, including rat, monkey and human.
Unbiased stereological methods were used for estimating the numerical density and the total number of claustral neurones projecting to the cingulate cortex in rabbit and rat. In rat the numerical density of neurones projecting to the retrosplenial granular cortex (RSG) differed significantly from those projecting to the retrosplenial agranular (RSA) and cingulate (Cg) cortices while in rabbit the numerical densities of retrogradely labelled neurones in the claustrum following injections into various areas of the cerebral cortex did not differ significantly. The total number of retrogradely labelled neurones in the claustral limbic zones did not differ significantly in both species. The quantitative analysis of claustral zones projecting to a different cingulate cortex area, both in rabbit and rat, reveals that each of these zones is rather homogeneous.
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