Immunohistochemical demonstration of specific antigens in the human brain fixed in zinc-ethanol-formaldehyde

Коржевский, Д. Э.; Sukhorukova, E. G.; Кирик, О. В.; Grigor’ev, I. P.

doi:10.4081/ejh.2015.2530

Cited by 61 publications

(23 citation statements)

References 27 publications

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“…The material was fixed in zinc-ethanol-formaldehyde [ 17 ] and immersed into paraffin. Paraffin blocks were used to prepare slices with a thickness of 5, 7, and 10 μm, which were pasted onto slides with an adhesive coating (Histobond, Polysine, SuperFrost Gold, Germany).…”

Section: Methodsmentioning

confidence: 99%

The Spatial Organization of the Intranuclear Structures of Human Brain Dopaminergic Neurons

Коржевский¹,

Гусельникова²,

Кирик³

et al. 2017

Acta Naturae

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We studied the intranuclear localization of protein nucleophosmin (B23) and ubiquitin in the dopaminergic neurons of human substantia nigra (n = 6, age of 25–87 years) using immunohistochemistry and confocal laser microscopy. Intranuclear ubiquitin-immunopositive bodies that morphologically correspond to Marinesco bodies were found to be present in substantia nigra dopaminergic (tyrosine hydroxylase-immunopositive) neurons but absent in non-dopaminergic neurons. The number of bodies varied from 0 to 6 per cell nucleus. Nucleophosmin (B23) was found in the neuronal nucleolus, with the nucleolus size being constant in the nigral neurons of each individual brain. All the observed neurons had only one large nucleolus with intense nucleophosmin immunoreactivity and a lightly stained region (1–2 μm in diameter) that apparently represents the giant fibrillar center (GFC). An intensely immunostained nucleophosmin-containing granule was often observed at the GFC periphery. Double labeling demonstrated that nucleophosmin-immunoreactive nucleolus and ubiquitin-immunoreactive Marinesco bodies can occur both closely to and remotely from each other. Three-dimensional reconstruction indicates that rounded Marinesco bodies are polymorphic and often have a complex shape, with some flattening and concavities, which may be associated with contact not only with the nucleolus, but also, presumably, with other intranuclear structures free of ubiquitin or nucleophosmin. Ubiquitin-immunoreactive structures with a relatively small size (up to 1 μm in length) and various clastosome-like shapes (Lafarga et al., 2002) often occur near Marinesco bodies. There were no cases of detection of ubiquitin in the nucleoli of dopaminergic neurons and nucleophosmin/B23 in typical Marinesco bodies. The obtained information may be helpful in unraveling the molecular mechanisms of the selective vulnerability of substantia nigra dopaminergic neurons to damaging factors.

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Section: Methodsmentioning

confidence: 99%

The Spatial Organization of the Intranuclear Structures of Human Brain Dopaminergic Neurons

Коржевский¹,

Гусельникова²,

Кирик³

et al. 2017

Acta Naturae

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“…В настоящее время производится широкий спектр моноклональных антител к ТГ, а также поликлональных антител, которые позволяют выявлять этот фермент иммуногистохимически в нервной системе и других отделах организма человека и различных видов животных. В наших предыдущих исследованиях было показано, что [24,25].…”

Section: Immunohistochemical Markers For Neurobiologyunclassified

“…Показано, что высокое качество окраски парафиновых срезов тканей можно достичь при использовании протокола окраски, предложенного нами ранее [4]. Допустимы различные виды фиксации гистологического материала, однако оптимальные результаты окрашивания были получены при использовании цинк-этанол-формальдегида [24,25]. С помощью данного метода было изучено распределение холинергических нервных клеток спинного мозга (СМ) крысы на разных сроках эмбрионального развития, в раннем постнатальном периоде и у взрослых животных [3,40,41].…”

Section: Immunohistochemical Markers For Neurobiologyunclassified

“…В наших исследованиях применяли моноклональные мышиные антитела к белку SMI-32 (Bio Legend, США) для исследования нервных волокон седалищного нерва крысы в норме и после травмы. Реакцию проводили на парафиновых срезах после фиксации материала в цинк-этанол-формальдегиде [24,25].…”

Section: антитела Smi-32 против белков нейрофиламентовunclassified

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Immunohistochemical markers for neurobiology

Коржевский

Эдуардович²,

Grigor’ev

et al. 2020

Medical academic journal

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In neurobiological studies, crucial is the selection of most appropriate and informative experimental methods, one of which is immunohistochemistry. This review briefly summarizes the experience of adaptation of immunohistochemical methods to nervous system studies accumulated over years the Laboratory of Functional Morphology of the Central and Peripheral Nervous System (Institute of Experimental Medicine). The aim of this work was to determine the most effective and reliable immunomarkers for neurobiological studies. The article contains theoretical basis and practical recommendations for use of key cytospecific and functional markers used in studies of structural and functional organization of brain and spinal cord of mammalian animals and human. In particular, the results of immunohistochemical reactions to neural markers (NeuN, neurofilament proteins, alpha-tubulin, alpha-synuclein), neurotransmitter synthesizing enzymes (tyrosine hydroxylase, glutamate decarboxylase, choline acetyltransferase, NO synthase) and glial markers (GFAP, glutamine synthetase, Iba-1, vimentin) are demonstrated. The presented methodology is useful for experimental neurobiology and clinical morphological diagnostics.

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“…In the Laboratory of Morphology at the Institute of Experimental Medicine (Saint Petersburg), a new zinc-containing combination fixative -zinc-ethanolformaldehyde (1 g of zinc chloride in 96% ethanol and concentrated formalin, 9:1) -was developed [84]; that reagent demonstrated good preservation of the brain tissue and internal organs of humans and laboratory animals [85,86]. Immunohistochemical study of nervous tissue (definitive and embryonic) and peripheral organs fixed in zinc-ethanol-formaldehyde allowed for detecting a large number of proteins: calbindin, calretinin, choline acetyltransferase, glutamate decarboxylase, glial fibrillary acidic protein, Iba-1 and NeuN proteins, neuronspecific enolase, synaptophysin, tyrosine hydroxylase, alpha-tubulin, vimentin, nestin and others [87][88][89][90][91][92][93][94][95][96][97][98].…”

Section: Reviewsmentioning

confidence: 99%

Current Technologies for Fixation of Biological Material for Immunohistochemical Analysis (Review)

Grigor’ev¹,

Коржевский²

2018

Sovrem Tehnol Med

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Based on their own experience and published reports the authors provide an insight into the existing methods of fixation of biological material used in immunohistochemistry. The first quality of an immunohistochemical fixative should be its ability to preserve the tissue structure so that the antigenic properties of macromolecules are minimally affected. Considering this point, the review analyzes the applicability of commonly used fixatives to immunohistochemical staining; among those, aldehydes (formaldehyde, glutaraldehyde, glyoxal), dehydrating (coagulating) agents (ethanol, methanol, acetone), combined fixation solutions (Bouin's solution, Carnoy's solution, methacarn, etc.), as well as the recent zinc-containing fixatives and commercial products. Most of these fixatives inevitably change the tertiary and quaternary structure of many proteins; therefore, the detection of these proteins by immunohistochemistry requires an additional procedure of unmasking the epitopes using proteolytic enzymes or elevated temperatures. When compared for the preservation of antigenic structures, a high quality of the novel zinc-containing fixative-zinc-ethanol-formaldehyde-was noted. It has been concluded that none of the fixatives known to date has such a combination of properties that allow obtaining high-quality histological preparations and, at the same time, allows for detecting of any antigens in the stained tissue.

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Immunohistochemical demonstration of specific antigens in the human brain fixed in zinc-ethanol-formaldehyde

Cited by 61 publications

References 27 publications

The Spatial Organization of the Intranuclear Structures of Human Brain Dopaminergic Neurons

The Spatial Organization of the Intranuclear Structures of Human Brain Dopaminergic Neurons

Immunohistochemical markers for neurobiology

Current Technologies for Fixation of Biological Material for Immunohistochemical Analysis (Review)

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