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1. Distribution of c-Fos positive nuclei in: ventral tegmental area, substantia nigra, periaqueductal gray. 2. Appearance of calbindin-D28k, calretinin and parvalbumin in midbrain neurons that are engaged in the stress response. 3. Changes of c-Fos and calcium-binding proteins expression during maturation. The material consisted of Wistar rats of age between 0 and 90 days. The OF exposure was applied throughout 10 min and 90 min before the death of the animals. The brain sections were double stained using the antibodies against c-Fos, CB, CR or PV. Our results showed that in all studied nuclei age-related increase of c-Fos expression (without changing of its distribution properties) was found. PV didn't show any co-localization with c-Fos in neurons of studied regions at any ages, however some PV-immunoreactive (PV-ir) basket-like structures around c-Fos-immunoreactive (c-Fos-ir) neurons were observed. In the youngest group of rats c-Fos-ir cells and cells immunoreactive for CB and CR constituted separate neuronal populations. During maturation increases in the level of their co-localization with c-Fos was observed. We may conclude that in adult rat midbrain structures CB-immunoreactive (CB-ir) and CR-immunoreactive (CR-ir) cells (probably projection neurons) are mainly activated in the stress response following OF exposure. In the contrary PV-ir cells has only an indirect (modulatory) influence upon the c-Fos-ir cells.
The purpose of this study was to trace the immunoreactivity of the two calcium binding proteins, parvalbumin and calbindin D28k, in the periaqueductal gray matter of the chinchilla midbrain. The immunoreactivity of these proteins in this species has never been investigated. The localisation of the activity was examined by carrying out the peroxidase-antiperoxidase (PAP) reaction using specific antibodies against parvalbumin and calbindin D28k. Slightly different parvalbumin immunoreactivity was shown. In most neurons, with the exception of large neurons in the dorsal and dorso-lateral periaqueductal gray matter, a weak immunostaining for parvalbumin was observed. Extremely intense immunostaining for calbindin D28k occurred in all neurons in the examined area. The results obtained suggest a slightly different distribution of parvalbumin in the neurons of the periaqueductal gray matter of the midbrain of the chinchilla than in other animal species. This indicates that mainly calbindin D28k is involved in the regulation of intracellular calcium ion concentration in the periaqueductal gray matter of the chinchilla.
Piracetam is clinically being used nootropic drug but the details of its neuroprotective mechanism are not well studied. The present study was conducted to assess the effects of piracetam on rotenone induced oxidative stress by using both ex vivo and in vivo test systems. Rats were treated with piracetam (600 mg/kg b.w. oral) for seven constitutive days prior to rotenone administration (intracerebroventricular, 12µg) in rat brain. Rotenone induced oxidative stress was assessed after 1 h and 24 h of rotenone administration. Ex vivo estimations were performed by using two experimental designs. In one experimental design the rat brain homogenate was treated with rotenone (1 mM, 2 mM and 4 mM) and rotenone+piracetam (10 mM) for 1 h. While in second experimental design the rats were pretreated with piracetam for seven consecutive days. On eighth day the rats were sacrificed, brain homogenate was prepared and treated with rotenone (1 mM, 2 mM and 4 mM) for 1h. After treatment the glutathione (GSH) and malondialdehyde (MDA) levels were estimated in brain homogenate. In vivo study showed that pretreatment of piracetam offered significant protection against rotenone induced decreased GSH and increased MDA level though the protection was region specific. But the co-treatment of piracetam with rotenone did not offer significant protection against rotenone induced oxidative stress in ex vivo study. Whereas ex vivo experiments in rat brain homogenate of piracetam pretreated rats, showed the significant protection against rotenone induced oxidative stress. Findings indicated that pretreatment of piracetam significantly attenuated the rotenone induced oxidative stress though the protection was region specific. Piracetam treatment to rats led to its absorption and accumulation in different brain regions as assessed by liquid chromatography mass spectrometry/mass spectrometry. In conclusion, study indicates the piracetam is able to enhance the antioxidant capacity in brain cells in region specific manner. The study is also revealing the rationale for its clinical use in cognitive impairment and other neurological diseases.
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
Jurkowlaniec E., Trojniar W. and Tokarski J.: The EEG activity after lesions of the diencephalic part of the zona incerta in rats, Acta Physiol. Pol. Neocortical and hippocampal EEG activity was recorded in 23 rats subjected to the bilateral electrolytic lesions of the diencephalic zona incerta (ZI). The aim was to find whether damage to ZI can replicate insomnia and disturbances in cortical EEG desynchronization and hippocampal theta rhythm found after lesions of the lateral hypothalamic (LH) area. No effect of the ZI lesions on waking-sleep cycle was found. The amplitude and frequency of cortical waves and hippocampal theta rhythm during waking were changed only in some rats. These changes were small, short-lasting and bidirectional (toward and increase or decrease in different subjects). Both the amplitude and frequency of paradoxical sleep theta were depressed in part of animals. Thus the marked EEG changes after LH lesions can not be attributed to simultaneous damage of the adjacent subthalamic region. However, the ZI seems to constitute a part of a larger system regulating cortical arousal and hippocampal theta rhythm.
The localisation of carletinin in the midbrains of 10 sexually mature chinchilla males was examined by carrying out the peroxidase-antiperoxidase immunocytochemical reaction using specific monoclonal antibody against calretinin. Intensive immunostaining for the protein was observed in the majority of fusiform, pyramidal, and stellate neurons of dorsal raphe nucleus. A similar calretinin distribution in neurons of this region to those observed in primates and rodents was demonstrated.
Glial cell line-derived neurotrophic factor (GDNF) was initially discovered as a neurotrophic factor that enhances survival of midbrain dopaminergic neurons. Findings in 1994 and 1995 extended the spectrum of biological activities of GDNF to different populations of neurons (motor and sensory). Recent findings revealed that GDNF, Neurturin (NTN), Persephin (PSP), and Artemin (ARTN) are members of the GDNF protein family and are structurally related to transforming growth factor protein family. They are survival factors for peripheral and central neurons, for oligodendrocytes and can promote morphogenesis of kidney in vitro.
Investigations were made on serial sections of human embryos at developmental stages 16 and 17 (37-41 days). The oculomotor nucleus in embryos at these stages is located at the cephalic flexure of the neural tube. It consists of paired lateral nucleus which is formed by well defined group of large neurons in the basal plate of the midbrain. Within the lateral nucleus two groups of cells (medial and lateral) can he distinguished. Posteriorly these groups unite and form a single nucleus.
Using stained acryl latex injected techniques the arteries of the base of the brain in the Mongolian gerbil (Meriones unguiculatus) were studied. The brain of the Mongolian gerbil is supplied with blood by even internal carotid arteries and odd basilar artery. The branches of the internal carotid arteries supply the forebrain and interbrain. They form the arterial circle at the base of the brain. The basilar artery is a source of blood for the midbrain and hindbrain. In this species, except one case, the opened type at the back side of arterial circle was found. The posterior communicating arteries between basilar artery and internal carotid arteries were not observed.
We studied the influence of Withania somnifera (Ws) root extract (100 mg/kg body weight) on parkinsonism induced by 1-methyl 4-phenyl 1,2,3,6-tetrahydropyridine (MPTP; i.p, 20 mg/kg body weight for 4 days), via the analysis of behavioral features and the oxidant-antioxidant imbalance in the midbrain of mice. A significant alteration in behavior, increased levels of thiobarbituric acid reactive substance (TBARS), and increased activities of superoxide dismutase (SOD) and catalase (CAT) were noticed in this region of brain in MPTP-treated mice. Oral treatment with the root extract resulted in a significant improvement in the mice’s behavoiur and antioxidant status, along with a significant reduction in the level of lipid peroxidation. The results indicated that at least part of the chronic stress-induced pathology may be due to oxidative stress, which is mitigated by Ws. Further studies are needed to assess the precise mechanism to support the clinical use of the plant as an antiparkinsonic drug.
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