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ABSTRACT: The distribution of the heme-containing protein, neuroglobin, in the rat forebrain was examined using immunohistochemistry in normal rats and in rats 48 h after transitory ischaemia induced by endovascular occlusion of the middle cerebral artery for 30 min. In rats with transitory ischaemia, an uneven distribution of neuroglobin was observed in the forebrain: maximal neuroglobin immunoreactivity was found in the cortex, caudate nucleus, choroid plexus, and ependymal lining, whereas the least neuroglobin immunoreactivity occurred in neural pathways. Unilateral brain ischaemia results in a profound decrease in neuroglobin immunoreactivity in the lesioned hemisphere. No increased expression of neuroglobin was detected in the penumbra or at points located distal from the lesioned site.
Key words: brain, neuroglobin, immunohistochemistry, ischaemia.
Journal of Asymmetry. 04/2013; 7(1):15-21.
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ABSTRACT: Ependymal cells of cerebral ventricles are the most probable candidate progenitor cells. We studied ependymal and subventricular zone cells expressing intermediate filament protein vimentin. The results suggest that the ventricular ependyma represent a homogenous cell population. Some ependymocytes have long processes contacting with blood vessels, which makes them similar to tanycytes of the third cerebral ventricle. New data on the structure of ependymocyte processes attest to their active involvement into creation of a special microenvironment for neural stem cells of the subventricular proliferative zone.
Bulletin of Experimental Biology and Medicine 01/2013; 154(4):553-7. · 0.27 Impact Factor
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ABSTRACT: Forebrain subventricular zone (SVZ)--the putative major source of neural stem cells in the brain of adult mammals--can hardly be visualized using routine histological staining. The present study was focused on the possibility of application of immunocytochemical approach for accurate delineation of the border between SVZ and striatum. It was shown that immunocytochemical reactions demonstrating tyrosine hydroxylase or synaptophysin were optimal for the determination of the border between SVZ and striatum in different mammals.
Morfologiia (Saint Petersburg, Russia) 01/2012; 141(1):81-4.
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ABSTRACT: Microgliocytes are currently known as structurally labile cells, that, by changing the length of their processes, control the synaptic organization of the brain. The aim of this study was to investigate structural organization of microglia of the striatum and of the forebrain periventricular area in rat following transitory focal ischemia. In the absence of brain infarction, ischemic lesion was found to activate microglia, which contributed to rearrangement of striatal neuropil. As microgliocytes became activated, they changed their shape from characteristic dendritic one to oval and rounded, which are typical to amoeboid microglia. Microgliocytes were shown to be capable of proliferation.
Morfologiia (Saint Petersburg, Russia) 01/2012; 141(2):28-32.
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ABSTRACT: Semiconductor nanocrystals, i.e., quantum dots (QD) are a new and promising (in terms of use in biology and, medicine) class of inorganic fluorescent materials. The aim of the present work was to identify the advantages and disadvantages of QD in immunocytochemical studies. Streptavidin-QD conjugates were found to have a series of advantages over streptavidin conjugates with organic fluorochromes (high fluorescence intensity, photostability, wider range of excitation, narrow and symmetrical emission peak), which allows them to be recommended for use in immunocytochemical studies. However, a number of disadvantages were also identified (lower stability on prolonged storage than conjugates with organic fluorochromes, poor preservation of aliquots of reagent, inability to retain fluorescence in stained specimens on prolonged storage, incompatibility with a number of commercially available media used for embedding specimens), preventing the wide use of nanocrystals in immunocytochemistry.
Neuroscience and Behavioral Physiology 10/2011;
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ABSTRACT: Calcium-binding protein Iba-1 is currently believed to be identical to AIF-1 protein and is regarded as a selective marker for microglia. However, data on populations of cells able to express Iba-1/AIF-1 are contradictory. The aim of the present work was to identify rat brain cells expressing Iba-1 protein and to describe their functional characteristics. Immunocytochemical and confocal laser microscopy methods were used. Iba-1 protein was seen only in microglial cells, meningeal macrophages, supraependymal macrophages, and superficial and stromal cells of the vascular plexus, i.e., cells with phagocytic functions. Comparison of these data with results from other studies suggests that not all of the currently used antibodies to Iba-1/AIF-1 protein recognize the same product. This may be associated with alternative splicing of the Iba-1 mRNA.
Neuroscience and Behavioral Physiology 02/2011; 41(2):149-52.
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ABSTRACT: This paper is aimed at the characterization of beta-III-tubulin, MAP2 and doublecortin, the proteins which participate in the organization, stabilization and function of the microtubules of nerve cell cytoskeleton. Due to the structural and functional features, these proteins may be regarded as differentiation markers, associated with neurogenesis and as the indicators of nerve cell functional status under normal and pathological conditions. The data presented show that these proteins perform important structural and transport functions in nerve cells and are essential for some neurospecific intracellular processes. However, current knowledge of the functional role of these proteins in nerve cells is insufficient and requires significant supplementation indispensable for unequivocal interpretation of the studies results.
Morfologiia (Saint Petersburg, Russia) 01/2011; 139(1):13-21.
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ABSTRACT: A study of structural and functional organization of the boundaries separating CNS compartments is a fundamental task of neurobiology. Taking into account the contradictory data on the structure of superficial layers of mammalian neocortex, it is pertinent to study structural and cytochemical organization of astrocytes--the main components of the brain barrier system in animals that are often used for experimental modeling of brain diseases and injuries. The aim of the present work was to study the structural organization of layer I astrocytes of rat neocortex. Astrocytes were demonstrated immunocytochemically using anti-GFAP, anti-vimentin and anti-nestin antibodies using light and confocal laser microscopy. The results of the study demonstrated that the superficial glial limiting membrane had significant structural differences in different cortical regions. Astrocytes in layer I of rat neocortex were different from typical protoplasmic astrocytes, common to gray matter The regional peculiar features of superficial glial limiting membrane organization that were found in this study, are probably determined by the differences in functional characteristics of CSF-encephalic barrier in the specific regions of the brain.
Morfologiia (Saint Petersburg, Russia) 01/2011; 140(6):32-5.
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ABSTRACT: Ezrin (cytovillin or p81 protein) is an actin-binding protein, a member of ERM (ezrin, radixin and moesin) family, which species contribute to stabilization of the plasma membrane-formed structures. The aim of the present study was to demonstrate the ezrin-containing cells in the rat brain and to describe their topography and morphological features. The most pronounced immunohistochemical reaction to ezrin was found in the epithelium of the choroid plexus, cells of the subcommissural organ and ventricular ependyma. Moreover, ezrin staining was also detected in the unidentifiable cells in the subventricular zone, rostral migration pathway and astrocytes in various brain areas. Preferential ezrin localization in the brain cells contributing to formation of barrier structures suggests its involvement in transport processes in the CNS.
Morfologiia (Saint Petersburg, Russia) 01/2011; 140(6):86-8.
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ABSTRACT: The aim of the study was to examine the distribution of cells expressing Msi-1 (Musashi-1) protein which is believed to be a marker of neural stem cells, in rat telencephalon. These cells were found to be concentrated in the subventricular proliferative zone and diffusely scattered in striatum and hippocampus. Moreover, neurons with extensive Msi-1 reaction were found in habenular nuclei. The data obtained are not in full agreement with current views on the localization of neural stem cells in the brain.
Morfologiia (Saint Petersburg, Russia) 01/2011; 139(2):77-9.
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ABSTRACT: Subventricular proliferative zone is the brain area most intriguing and least understood in terms of cellular and spatial organization. The objective of this study was to focus on the structural and cytochemical organization of astrocytes of the subventricular zone of the rat brain. Astrocytes were detected immunocytochemically. The preparations were analyzed using conventional light microscopy and confocal laser microscopy. The study performed showed that the major cellular population of the subventricular zone was represented by a special kind of atypical fibrous astrocytes with the intermediate filaments containing only glial fibrillary acidic protein (GFAP). The processes of these cells formed a complex double-layer network beneath the ependyma. The described structural pattern of the glial skeleton of the subventricular zone seems to be determined by specific functions of the area and its location in the zone of CSF-brain barrier.
Morfologiia (Saint Petersburg, Russia) 01/2011; 139(3):77-9.
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ABSTRACT: We developed a well reproducible method for detection of brain microglia on paraffin sections on the basis of modern immunohistochemical methods. Polyclonal antibodies to Iba1 antigen obtained after immunization with a synthetic peptide corresponding to 81-93 amino acid fragment of Iba1 antigen selectively interact with microglia cells on paraffin sections of rat brain. Detailed protocol of preparation treatment was drawn up ensuring complete detection of the microglia expressing Iba1 protein.
Bulletin of Experimental Biology and Medicine 11/2010; 149(6):768-70. · 0.27 Impact Factor
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ABSTRACT: In immunomorphological studies, it is often necessary to visualize several antigens simultaneously within the same histological sections. However in practice, the application of double staining is limited due to methodological complexity. The aim of the current study was to reproduce different methods recommended for multiple immunohistochemical staining of one section and to estimate their advantages and drawbacks. Six different methodological approaches for detection of several antigens in one slide were examined. It was found that the method providing the widest resources for analysis was the fluorescent detection, the most convenient was the application of ready-to-use standardized reagents, and the cheapest method was two-color reaction with DAB-type chromogen.
Morfologiia (Saint Petersburg, Russia) 01/2010; 137(5):59-64.
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ABSTRACT: Semi-conducting nanocrystals represent a new class of fluorescent inorganic objects which have a promising perspective for the application in biology and medicine. The aim of the current work was a determination of advantages and shortcimingss of quantum dots (QD) application in immunocytochemistry. It was shown that streptavidin-QD conjugates have more advantages (i.e. high intensity of fluorescence, photostability, wide excitation range, short and symmetric emission range) as compared to streptavidin conjugates with organic fluorochromes. This allows to recommend the use of QD in immunocytochemical studies. However, there are several disadvantages (like lower stability during long-term storage as compared to that one of organic fluorochrome conjugates, poor safety of aliquots, impossibility of long-term preservation of fluorescence of stained sections during their storage, incompatibility with several commercial mounting media) that limit the widespread application of nanocrystals in immunocytochemical studies.
Morfologiia (Saint Petersburg, Russia) 01/2010; 137(3):71-5.
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ABSTRACT: In vitro and in vivo studies addressing the differentiation of neural stem and progenitor cells in the CNS require the use of highly specific markers for neurons and gliocytes. The aim of the present work was to study the distribution of a marker for differentiating neurons, i.e., doublecortin (DCX), in structures of the brain and spinal cord in rat embryos during the period preceding the formation of the cortical plate using immunocytochemical methods and light and confocal microscopy. DCX was detected in three types of cell in the developing nervous system at 13-14 days of embryogenesis: neurons giving reactions for the nuclear marker for differentiated nerve cells NeuN, migrating and differentiating neuroblasts, and some cells which are members of the population of radial gliocytes. The quite high selectivity of DCX expression allows use of this marker to be recommended for studies of the early stages of nervous system development in mammals.
Neuroscience and Behavioral Physiology 08/2009; 39(6):513-6.
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ABSTRACT: Nestin is present in podocytes of the renal corpuscle in rats and humans. Specific differences manifested in more intensive and widespread expression of nestin by endothelial cells of blood vessels in human kidney.
Bulletin of Experimental Biology and Medicine 05/2009; 147(4):539-41. · 0.27 Impact Factor
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ABSTRACT: The reproducibility of brain injury was evaluated by simulating ischemia in rats by 30-min occlusion of the middle cerebral artery. The selected ischemia-reperfusion protocol was characterized by high reproduction of the striatal neuron injury, which fact suggests this model for studies of nerve tissue reactions to injury and for evaluation of the efficiency of neuroprotective drugs.
Bulletin of Experimental Biology and Medicine 03/2009; 147(2):255-6. · 0.27 Impact Factor
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ABSTRACT: Nestin, an intermediate filament protein, is known to be expressed in proliferating and provisional cells in the forming mammalian brain, disappearing on differentiation. The aim of the present work was to identify the morphological types and locations of cells regaining the ability to synthesize nestin after transient total brain ischemia in rats. Transient ischemia was found to be followed by the induction of nestin synthesis in astrocytes in the damaged area; these cells acquired structural features not characteristic of the adult brain, and these persisted in the long term. Nestin synthesis was also induced in proliferation-capable undifferentiated cells in the subventricular zone. The acquisition by astrocytes of some of the phenotypic features of immature glial cells, however, does not provide grounds for the notion that they were transformed into neural stem cells.
Neuroscience and Behavioral Physiology 03/2008; 38(2):139-43.
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ABSTRACT: The aim of the study was to detect the cells synthesizing intermediate filament protein vimentin in the telencephalon of intact rats and animals exposed to unilateral permanent occlusion of middle cerebral artery accepted as a model of ischemic stroke. In intact rat telencephalon, vimentin-positive cells were detected within the cerebral barriers. 14 days following the occlusion of middle cerebral artery, numerous vimentin-positive cells appeared in the perifocal zone of the lesion (penumbra) making up a majority of cells of the regenerating nervous tissue at the border with intact brain tissue. In the subependimal proliferative zone, a large number of small, vimentin-negative cells was observed that were located among the long processes of vimentin-positive cells directed towards the lesion area. The data obtained demonstrate predominant localization of vimentin-immunopositive cells of (in both intact and lesioned brain) in the zone of barrier structures' formation.
Morfologiia (Saint Petersburg, Russia) 02/2007; 132(5):23-7.
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ABSTRACT: Nestin, an intermediate filament protein, is known to be expressed in the proliferating and provisional cells of the developing mammalian brain and is lost during differentiation. The aim of the present study was to determine the morphological type and localization areas of the rat brain cells exhibiting the ability to synthesize nestin after a short-term global ischemia of the brain. Induction of nestin synthesis after a short-term ischemia was found within the injured brain areas in astrocytes, which exhibited structural features atypical for these cells in mature brain and maintained them for a long time, and in the morphologically undifferentiated cells of subventricular zone, which were able to proliferate. However, acquisition by astrocytes of some phenotypic properties of immature glial cell does not by itself support the view that they may transform into neural stem cells.
Morfologiia (Saint Petersburg, Russia) 02/2007; 131(1):23-6.
