Immunohistochemical markers for neurobiology

Abstract

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.

Experimental and Clinical Biomarkers for Progressive Evaluation of Neuropathology and Therapeutic Interventions for Acute and Chronic Neurological Disorders

“…There is variety in the distribution of GAD65 and GAD67 within the GABAergic neurons. GAD67 is more abundant in the cell bodies, processes, and synaptic terminals of neurons, whereas GAD65 is highly enriched in axon terminals and is associated with synaptic vesicles [ 157 ]. Hence, GAD67 is more often used as an immunohistochemical biomarker for GABAergic brain structures.…”

“…GAD67 is expressed in neurons of the hippocampus, cortex, cerebellum, olfactory bulb, striatum, paleostriatum, reticular zone of the substantia nigra, and inferior colliculus. Various neurological and psychiatric diseases such as ischemia, schizophrenia, and PD can impair GAD67 expression [ 157 ]. The formation of tau-containing extracellular vesicles (EVs), which is a part of AD, is facilitated by GAD67 [ 159 ].…”

“…TH is an enzyme involved in the conversion of the amino acid L-tyrosine to the precursor for dopamine, L-3,4-dihydroxyphenylalanine (L-DOPA). It is the rate-limiting enzyme involved in the synthesis of catecholamines, such as dopamine, noradrenaline, and adrenaline [ 157 ]. These neurotransmitters regulate a variety of physiological and psychoemotional functions and reactions, including stress, sleep, learning, memory, attention, and energy metabolism.…”

“…It helps to identify patterns of neuroinflammation and subsequent damage [ 184 ]. The increased expression of GFAP has been detected in most CNS pathologies, such as TBI, ischemia, and neurodegenerative diseases [ 157 ]. A detailed methodology of GFAP histological staining is available in recent papers [ 43 , 144 ].…”

“…Vimentin is a highly conserved protein (57 kDa) found in tissues of vertebrates, especially in mesenchymal cells [ 157 ]. Vimentin belongs to a class III type IF and is considered the only IF type expressed in a range of cells, including astrocytes, fibroblasts, endothelial cells, neutrophils, lymphocytes, and macrophages [ 178 ].…”