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The present study investigates the development of microglial and astroglial cells in the postnatal rat striatum, using immunohistochemical methods with panel antibodies that recognize macrophage antigens of unknown function - ED l, complement type 3 receptor- OX-42 (for microglia) and glial fibrillary acidic protein (for astrocytes). On the day of birth, EDl/OX-42- immunoreactive microglial cells present in the striatum represent ameboid microglia. Between PO and P10 we could observe the migration of ameboid microglial cells from neuroepithelial ventricular zone through internal and external capsules into the striatum. During the second postnatal week (PIO, P14) a considerable decline of ameboid EDl-immunoreactive microglial cells and an increase of the number of OX-42 positive ramified cells were observed. At P21 only OX-42 positive ramified cells were observed in the whole striatum. On the day of birth, only a few GFAP positive cells resembling radial glia were observed in the striatium. During the first postnatal week, the number of GFAP-positive cells increased significantly; they showed typical morphology of the astrocytes present in the adult animals. After P22 the final striatal population of astroglia was formed.
We examined the expression of brain nitric oxide synthase (bNOS) in two developing rat brain structures, the striatum and the cerebral cortex. For this purpose, we quantified the relative protein concentration level using the Western blotting method and densitometric scanning. 32 Wistar rats, divided according to survival period (P0-P120-postnatal days) were used in this study. Our results demonstrate that bNOS expression rises in these structures during the first week of postnatal life, reaching a maximum in the striatum on the 10th day and in the cerebral cortex on the 7th day of postnatal life. After the period of increase the expression declines and after the 14th day a stabilisation of bone protein concentration is observed, both in the striatum and the cerebral cortex. These changes in bone protein expression might be related to the important role of nitric oxide in the developing rat brain, especially in synaptogenesis, apoptosis and neurotransmission.
The developmental changes of 25-kDa synaptosomal-associated protein (SNAP-25) expression in the rat striatum and cerebral cortex were examined using Western-blotting and densitometric scanning of immunoblots. Analysis of the striatum extracts from postnatal day 0 (P0) to postnatal day 120 (P120) demonstrated that SNAP-25 is poorly expressed until P14. From this point the expression level gradually increases to reach a maximum on P60 and then decreases. The pattern of SNAP-25 expression in the rat cerebral cortex is different. Two peaks are observed, the first on P10 and the second on P60, after which the expression level decreases. These results appear to confirm the role of SNAP-25 protein in axon outgrowth and synaptogenesis in the nervous system.
Previous neuroimaging studies have shown that executive processes requiring planning and set-shifting [e.g. Montreal card sorting task (MCST)] may engage the dorsolateral prefrontal cortex (DLPFC) while inducing dopamine (DA) release in the striatum. However, functional imaging studies can only provide neuronal correlates of cognitive performance and cannot establish a causal relation between observed brain activity and task performance. In order to investigate the contribution of the DLPFC during set-shifting and its effect on the striatal DAergic system, we applied continuous theta burst stimulation (cTBS) to left and right DLPFC. Our aim was to transiently disrupt its function and to measure MCST performance and striatal DA release during [11C]raclopride PET. cTBS of the left DLPFC impaired MCST performance and DA release in the ipsilateral caudate–anterior putamen and contralateral caudate, as compared to cTBS of the vertex (control). These effects were limited only to left DLPFC stimulation but not right DLPFC. This is the fi rst study showing that cTBS, by disrupting left DLPFC function, may indirectly affect striatal DA release during performance of executive tasks. This cTBS-induced regional prefrontal effect and modulation of the frontostriatal network may be important for understanding the contribution of hemisphere laterality and its neural bases with regard to executive functions, as well as for revealing the neurochemical substrate underlying cognitive defi cits.
Organotypic slice cultures were established as a model that own properties of both cell culture and animal model. The most often used slice culture is derived from hippocampus but depending on the part of brain affected with pathology, researchers established cultures from cerebellum, midbrain or striatum. Above mentioned models allowed the investigation of disorders resulting from e.g. ischemia, trauma or toxic injury. Besides the brain injury, numerous studies were focused on spinal cord pathology connected with demyelination, inflammation or injury. Here, we describe the development of an in vitro model of longitudinal spinal cord slice culture. Compared to cell (neuron-oligodendrocyte) co-cultures, organotypic slices retain tissue organization as well as cell-cell contacts and therefore more closely mimic the environment in vivo. We demonstrate the applicability of this approach for xenograft transplantation of oligodendrocyte precursor cells derived from rat brain and mesenchymal stem cells derived from human umbilical cord. Stem cells fate after transplantation was observed in two paradigms: after cell transplantation on the top of spinal cord slice cultures (SCC) or cocultivation of cell culture with SCC space separated for 24 h. We observed the different morphology and protein expression of stem cells derived from different sources. Moreover, the same stem cells co-cultured with slices derived from different part of brain (hippocampus or spinal cord) expressed other markers. The method of longitudinal spinal cord slices enables observation of long fibers trajectory, new connections and neurorepair mechanisms. Moreover, it provides a time-efficient and costeffective adjunct to cell lines or in vivo transplantation models for study spinal cord pathology or experimental therapies. Furthermore, the approach can be readily used to assess the effect of pharmacological manipulations on myelin, providing a tool to better understand myelination and develop effective therapeutic strategies to treat myelin-related diseases. Supported by National Science Centre grant: 05728/B/NZ4/2011/01
5-HT2A/2C receptors are one of the most important in controlling basal ganglia outputs. In rodent models of Parkinson's disease (PD) blockade of these receptors increases locomotion and enhances the actions of dopamine (DA) replacement therapy. Moreover, previously we established that 5-HT2A/2C antagonist attenuate DA D1 agonist mediated vacuous chewing movements (VCMs) which are considered as an animal representation of human dyskinesia. These findings implicate 5-HT neuronal phenotypes in basal ganglia pathology, and promote 5-HT2 antagonists as a rational treatment approach for dyskinesia that is prominent in most instances of PD replacement therapy. In the current study we determined whether ketanserin (KET) and/or amphetamine (AMPH) affected dopaminergic neurotranssmision in intact and fully DA-denervated rats. Moreover, we looked into extraneuronal content of HO. of the neostriatum after AMPH and/or KET injection, assessed by HPLC analysis of dihydroxybenzoic acids (2,3- and 2, 5-DHBA) - spin trap products of salicylate. Findings from the present study demonstrated that there are no substantial differences in extraneuronal HO. generation in the neostriatum between control and parkinsonian rats. KET did not affect DA release in the fully DA-denervated rat's neostriatum and also did not enhance HO. production. As 5-HT2A/2C receptor-mediated transmission might prove usefulness not only in addressing motor complications of PD patients (dyskinesia) but also in addressing non-motor problems such depression and/or L-DOPA evoked psychosis, the findings from the current study showed that the use of 5-HT2A/2C receptor antagonists in Parkinson's disease does not impend the neostriatal neuropil to be damaged by these drugs. We concluded that 5-HT2A/2C receptor antagonists may provide an attractive non-dopaminergic target for improving therapies for some basal ganglia disorders.
Hepatic encephalopathy (HE) is characterized by motor symptoms associated with disturbed functions of the dopaminergic systems, but the underlying mechanisms are not clear. A previous study from our laboratories revealed that HE, induced in rats by repeated treatment with thioacetamide, enhanced the 50 mM potassium (KC1) -stimulated release of newly loaded [3H]dopamine in both striatal and frontal cerebral cortical slices in the presence of Ca2+. In the present study we compared the effects of HE on dopamine release in striatal and frontal cerebral cortical slices and synaptosomes in the presence and absence of Ca2+. HE enhanced the KCl-stimulated [3H]dopamine release from striatal and frontal cortical synaptosomes in the presence of Ca2+ to the same extent as in slices prepared from the respective brain regions. In the absence of Ca2+ a slight reduction in dopamine release was observed in frontal cortical synaptosomes from HE rats when compared to control rats, while no effect of HE on the release was discernible in frontal cortical and striatal slices and striatal synaptosomes. We conclude that in both brain regions studied HE stimulates dopamine exocytosis triggered by Ca2+ influx without affecting the release mediated by means of plasma membrane transporters or exocytosis involving intraterminal Ca2+.
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