[CURRENT HISTOCHEMICAL METHODS OF TISSUE IRON DEMONSTRATION BASED ON PERLS' REACTION].

The article presents the information on the modern modifications of Perls' reaction including diaminobenzidine enhancement according to R. Meguro (for various tissues). M.A. Smith (for nervous tissue), S.M. Levine (for detection of iron in oligodendrocytes and myelinated nervous fibers), and our own modification for visualization of the nucleolar iron. The analysis is performed of the advantages and disadvantages of these modifications of histochemical demonstration of iron. It is shown that the use of new methodological approaches significantly increases the sensitivity of Perls' reaction. Control procedures allow to to eliminate the possibility of artifacts.

[INTRANUCLEAR IRON DISTRIBUTION IN THE PURKINJE CELLS OF HUMAN CEREBELLUM].

The distribution of iron ions in the cerebellum of 15 human subjects aged 20-89-years was studied using highly-sensitive variant of Perls' histochemical technique. Increased iron content was found in the white matter and in Purkinje cells. In 10 out of 15 cases examined iron was detected in the nuclei of Purkinje cells, while in some cases iron was found in the nucleolus.

[Simultaneous demonstration of glutamate decarboxylase and synaptophysin in paraffin sections of rat cerebellum].

The article presents highly reproducible and inexpensive protocol for simultaneous demonstration of glutamate decarboxylase (GAD67), the key enzyme of gamma-aminobutyric acid (GABA) synthesis and synaptophysin (SYP), a marker protein of synaptic vesicles using confocal laser microscopy. In the cerebellar cortex, GAD labels Purkinje cells and pinceaux in their basal parts and is unevenly distributed in the neuropil of molecular and granular layers. SYP clearly marks the contours of large dendrites of Purkinje cells in molecular layer, while in the granular layers it labels parts of cerebellar glomeruli--the terminals of the mossy fibers. GAD-immunopositive structures (GABA-ergic axons of stellate cells--Golgi cells) are often located at periphery of the glomeruli. In the peripheral zone of the glomeruli, colocalization of GAD- and SYP-immunopositive structures was observed, suggesting the presence of GABA-ergic synapses in this zone.

[A synaptic marker synaptophysin].

The review summarizes the current data on synaptophysin (SYP), its functional role in the cell and the use of SYP immunocytochemistry for labeling the synaptic contacts. SYP is a transmembrane glycoprotein found in small presynaptic vesicles of the nerve cells and in microvesicles of the neuroendocrine cells. Literature data and the authors' own experience suggest that currently SYP is an important synaptic marker, which allows, with the use of light and confocal laser microscopy, to obtain the reliable data on the morphological organization of the synaptic structures in the central nervous system. It is also indispensable in the study of the efferent innervation of the internal organs. Applicatioin of the quantitative analysis of SYP-immunopositive structures using light and confocal laser microscopy allows to solve some problems that previously could be solved only by using electron microscopy.

[DISTRIBUTION OF THE MARINESCO BODIES IN THE NEURONS OF HUMAN BRAIN SUBSTANTIA NIGRA].

The aim of the study was to investigate the occurrence and intranuclear distribution of Marinesco bodies in substantia nigra neurons of the human brain. Marinesco bodies were identified in substantia nigra sections of 5 men aged 28 to 58 years old using Nissl staining and immunohistochemical detection of ubiquitin--the protein characteristic of this intranuclear inclusion. Marinesco bodies were found in 1-2% of the substantia nigra neurons, but not in adjacent brain areas. One neuron contained 1-4 Marinesco bodies sized up to 6.7x5.1 microm, which were located both near and at a distance from the nucleolus. Most Marinesco bodies exhibited ubiquitin expression. A trend was found for the increased incidence of Marinesco bodies in human substantia nigra neurons with age.

[Demonstration of the glomeruli in human cerebellum using synaptophysin immunohistochemistry and confocal laser microscopy].

The aim of the study was to develop the methodologic approaches for the demonstration of the complex synaptic groups (glomeruli) in human cerebellum using synaptophysin immunocytochemistry. The protocols presented in the paper allow to obtain of high qualify prepartaions for conventional light and confocal laser microscopy in which individual glomeruli and intraglomerular axonal terminals could be distinctly identified. The preparations obtained are suitable for a quantitative analysis.