scholarly journals A reliable method combining horseradish peroxidase histochemistry with immuno-beta-galactosidase staining.

1988 ◽  
Vol 36 (9) ◽  
pp. 1091-1096 ◽  
Author(s):  
M Sakanaka ◽  
S Magari ◽  
T Shibasaki ◽  
K Shinoda ◽  
J Kohno
Keyword(s):  
Cerebral Cortex ◽  
Horseradish Peroxidase ◽  
Pyramidal Cells ◽  
Nerve Fibers ◽  
Immunohistochemical Method ◽  
Double Labeling ◽  
Enzyme Systems ◽  
Pap Method ◽  
Hrp Histochemistry ◽  
Beta Galactosidase

A sensitive combination of horseradish peroxidase (HRP) tracing and immunohistochemistry was used by Rye et al. [J Histochem Cytochem (1984) 32:1145] in a search for the origins of neurotransmitter- and neuromodulator-containing nerve fibers in brain. In this combination, peroxidase as a marker in immunohistochemistry was thought to yield a homogeneous brown immunoreaction product of diaminobenzidine, different from the black granular reaction product of retrogradely transported HRP, which is visualized by the tetramethylbenzidine (TMB) reaction and subsequent stabilization. A neuron that exhibits both kinds of reaction products in its cytoplasm in sections subjected to combination staining is referred to as a double-labeled cell. With a combined HRP and corticotropin-releasing factor (CRF) immunoperoxidase-antiperoxidase (PAP) method, the first set of experiments showed "false" double-labeled cells in the pyramidal cell layer of rat cerebral cortex, but only rarely in the subcortical areas, possibly because of the use of one enzyme system in two different histochemical procedures. This limitation of the double-staining technique prompted us to demonstrate an alternate combination of HRP tracing and immunohistochemistry in the second set of experiments by employing two previously described independent enzyme systems: HRP as a retrograde tracer and beta-galactosidase as a marker for immunohistochemical demonstration of CRF. A homogeneous blue reaction product indicated immuno-beta-galactosidase staining, and a granular black or brown reaction product labeled retrogradely transported HRP in double-labeled cells in subcortical regions. Neither double labeling nor "false" double labeling was seen in pyramidal cells of cerebral cortex. These findings suggest that application of two independent enzyme systems in a combined HRP and immunohistochemical method may be useful for investigating in origins of peptidergic fibers in brain when the combination of HRP histochemistry and the PAP method appears to be inappropriate.

scholarly journals Cholinergic and VIPergic Innervation in Cerebral Arteries: A Sequential Double-Labeling Immunohistochemical Study

1990 ◽  
Vol 10 (1) ◽  
pp. 32-37 ◽  
Author(s):  
Frederick Jia-Pei Miao ◽  
Tony Jer-Fu Lee
Keyword(s):  
Vasoactive Intestinal Polypeptide ◽  
Arterial Wall ◽  
Immunohistochemical Study ◽  
Picric Acid ◽  
Cerebral Arteries ◽  
Nerve Fibers ◽  
Morphological Data ◽  
Immunohistochemical Method ◽  
Double Labeling ◽  
Functional Interactions

The possible co-localization of choline acetyltransferase (ChAT) and vasoactive intestinal polypeptide (VIP) in the nerve fibers of cat cerebral arteries was examined by a sequential double-labeling immunohistochemical method. Diaminobenzidine and tetramethylbenzidine were used as chromogens to distinguish ChAT (protein) and VIP (peptide) immunoreactivities. Since available fixatives often did not provide simultaneous preservation of optimal protein and peptide immunoreactivities, a new fixative, buffered periodate-paraformal-dehyde-picric acid-formaldehyde-lysine (PPPFL), was formulated and tested. PPPFL fixative is more reliable for simultaneously preserving ChAT and VIP immunoreactivities than were periodate-lysine-paraformaldehyde (PLP) fixative, Zamboni's fixative, or 2% paraformaldehyde solution alone. Using PPPFL as fixative, both ChAT immunoreactive (ChAT-I) and VIP-immunoreactive (VIP-I) fibers in cerebral arteries appeared as bundle and fine fibers. Most ChAT-I and VIP-I fibers were separate. Portions of ChAT-I and VIP-I fibers often ran closely in parallel or across each other. Overlaying of VIP-I on ChAT-I fibers and relay connections between them were also observed. These morphological data suggest the potential functional interactions between cholinergic and VIPergic innervations. In <5% of the fibers examined did ChAT and VIP immunoreactivities appear to be co-localized. These data therefore do not support the hypothesis that acetylcholine and VIP are co-localized in most fibers innervating the cerebral arterial wall.

C-fos, ERK1/2, MAP2, NOTCH1 Proteins Expression Patterns in Human Cerebral Cortex Neurons after Ischemic Stroke

2020 ◽  
Vol 75 (3) ◽  
pp. 226-233
Author(s):  
Svetlana P. Sergeeva ◽  
Aleksey V. Lyundup ◽  
Valery V. Beregovykh ◽  
Petr F. Litvitskiy ◽  
Aleksey A. Savin ◽  
...  
Keyword(s):  
Cerebral Cortex ◽  
Ischemic Stroke ◽  
Expression Patterns ◽  
Immunohistochemical Method ◽  
Left Middle Cerebral Artery ◽  
Contralateral Hemisphere ◽  
Human Cerebral Cortex ◽  
Fos Protein ◽  
Urgent Task ◽  
Indirect Immunoperoxidase

Background. The search for protein (these include c-fos, ERK1/2, MAP2, NOTCH1) expression that provide neuroplasticity mechanisms of the cerebral cortex after ischemic stroke (IS) patterns is an urgent task. Aims to reveal c-fos, ERK1/2, MAP2, NOTCH1 proteins expression patterns in human cerebral cortex neurons after IS. Materials and methods. We studied 9 left middle cerebral artery (LMCA) IS patients cerebral cortex samples from 3 zones: 1 the zone adjacent to the necrotic tissue focus; 2 zone remote from the previous one by 47 cm; 3 zone of the contralateral hemisphere, symmetric to the IS focus. Control samples were obtained from 3 accident died people. Identification of targeted proteins NSE, c-fos, ERK1/2, MAP2, NOTCH1 was performed by indirect immunoperoxidase immunohistochemical method. Results. Moving away from the ischemic focus, there is an increase in the density of neurons and a decrease in the damaged neurons proportion, the largest share of c-fos protein positive neurons in zone 2, NOTCH1 positive neurons in zone 1, smaller fractions of ERK1/2 and MAP2 positive neurons compared to the control only in samples of zone 1. Conclusions. With the IS development, the contralateral hemisphere is intact tissue increased activation zone, while the zones 1 and 2 have pathological activation signs. In zone 1 of the range, the adaptive response of the tissue decreases, and in zone 2 it expands. Therefore, a key target for therapeutic intervention is zone 2.

scholarly journals К. Schaffer: Zur feineren Struktur der Hirnrinde und über funktionelle Bedeutung der Nervenzellenfortsätze. Arch. f. Mikrosk. Anatomie. XLVIIL 4 Heft. 1897. S. S. 550—572

2020 ◽  
Vol V (3) ◽  
pp. 167-169
Author(s):  
A. E. Smirnov
Keyword(s):  
Cerebral Cortex ◽  
Opposite Direction ◽  
Molecular Layer ◽  
Pyramidal Cells ◽  
Special Group ◽  
The Brain ◽  
Axial Cylinder

The author's research refers to the anterior cerebral cortex of a newborn dog. The author studies in detail the so-called tiny pyramidal cells, lying between the pluripolar cells of the molecular layer and the small (true) pyramidal cells. Already R. y Cajal drew attention to polygonal or core-shaped cells, the cells that lie behind the layer of the outermost cells (pluripolare Nervenzellen von R. y Cajal), but did not separate them into a special group, believing that these cells were gradually changing vid, go into the small pyramids, to which he numbered them. Schaffer separates these cells into a special group, calling it the layer of surface polymorphic cells. These cells have a dark variety of shapes (fusiform, oval, roundish, pear-shaped, polygonal) and lie in approximately four (4) rows. Dendrites go then, mainly, in two opposite directions (for fusiform cells), then they move radially in all directions (for round and polygonal cells). The number of dendrites is sometimes strikingly abundant. Dendrites going to the surface of the brain reach it, while dendrites of the opposite direction sometimes go down to the ammonium formations of the cerebral cortex. Special attention should be paid to the axial cylinder of the disassembled cells; on the basis of the features of this appendix, the author distinguishes 3 types of disassembled cells.

A unitary hypothesis of mind-brain interaction in the cerebral cortex

1990 ◽  
Vol 240 (1299) ◽  
pp. 433-451 ◽  
Keyword(s):  
Cerebral Cortex ◽  
Pyramidal Cells ◽  
Neuronal Structure ◽  
Mental Experience ◽  
Effective Selection ◽  
Global Nature ◽  
The Mind ◽  
Mental Events ◽  
Apical Dendrites ◽  
The Brain

A brief introduction to the brain-mind problem leads on to a survey of the neuronal structure of the cerebral cortex. It is proposed that the basic receptive units are the bundles or clusters of apical dendrites of the pyramidal cells of laminae V and III-II as described by Fleischhauer and Peters and their associates. There are up to 100 apical dendrites in these receptive units, named dendrons. Each dendron would have an input of up to 100000 spine synapses. There are about 40 million dendrons in the human cerebral cortex. A study of the influence of mental events on the brain leads to the hypothesis that all mental events, the whole of the World 2 of Popper, are composed of mental units, each carrying its own characteristic mental experience. It is further proposed that each mental unit, named psychon, is uniquely linked to a dendron. So the mind-brain problem reduces to the interaction between a dendron and its psychon for all the 40 million linked units. In my 1986 paper ( Proc. R. Soc. Lond . B 227, 411-428) on the mind-brain problem, there was developed the concept that the operation of the synaptic microsites involved displacement of particles so small that they were within range of the uncertainty principle of Heisenberg. The psychon-dendron interaction provides a much improved basis for effective selection by a process analogous to a quantal probability field. In the fully developed hypothesis psychons act on dendrons in the whole world of conscious experiences and dendrons act on psychons in all perceptions and memories. It is shown how these interactions involve no violation of the conservation laws. There are great potentialities of these unitary concepts, for example as an explanation of the global nature of a visual experience from moment to moment. It would seem that there can be psychons not linked to dendrons, but only to other psychons, creating what we may call a psychon world.

scholarly journals LOCALIZATION OF α-FETOPROTEIN IN HUMAN AND RAT FETAL LIVERS AN IMMUNOHISTOCHEMICAL METHOD USING HORSERADISH PEROXIDASE

1974 ◽  
Vol 22 (6) ◽  
pp. 414-418 ◽  
Author(s):  
N. C. NAYAK ◽  
P. K. DAS ◽  
U. N. BHUYAN ◽  
ASHA MITTAL
Keyword(s):  
Connective Tissue ◽  
Horseradish Peroxidase ◽  
Immunohistochemical Method ◽  
Immunoperoxidase Technique ◽  
Serial Sections ◽  
Immunofluorescence Method ◽  
Immunohistochemical Technique

α-Fetoprotein (AFP) was successfully demonstrated in paraffin-embedded sections of human and rat fetal livers, by a multilayering immunohistochemical technique using the peroxidase-antiperoxidase system. Serial sections simultaneously subjected to immunofluorescence showed identical sites of localization in hepatocytic cytoplasm and sometimes in perivenous connective tissue. AFP-containing hepatocytes were located around efferent veins as well as randomly in the lobule. The immunoperoxidase technique has certain advantages over the immunofluorescence method which would justify the former's application in studies on the dynamics of AFP synthesis.

Cell lineage in the rat cerebral cortex: a study using retroviral-mediated gene transfer

Development ◽  
1988 ◽  
Vol 104 (3) ◽  
pp. 473-482 ◽  
Author(s):  
J. Price ◽  
L. Thurlow
Keyword(s):  
Cerebral Cortex ◽  
Progenitor Cells ◽  
Grey Matter ◽  
Cell Lineage ◽  
Cell Types ◽  
Retroviral Vector ◽  
Rat Cerebral Cortex ◽  
Rat Embryos ◽  
Bacterial Enzyme ◽  
Beta Galactosidase

We have used a retroviral vector that codes for the bacterial enzyme beta-galactosidase to study cell lineage in the rat cerebral cortex. This vector has been used to label progenitor cells in the cerebral cortices of rat embryos during the period of neurogenesis. When these embryos are allowed to develop to adulthood, the clones of cells derived from the marked progenitor cells can be identified histochemically. In this way, we can ask what are the lineage relationships between different neural cell types. From these studies, we conclude that there are two distinct types of progenitor cells in the developing cortex. One generates only grey matter astrocytes, whereas the second gives rise to neurones - both pyramidal and nonpyramidal - and to another class of cells that we have tentatively identified as glial cells of the white matter. We have also been able to address the question of how neurones are dispersed in the cortex during histogenesis. It had been previously hypothesized that clonally related neurones migrated radially to form columns in the mature cortex. However, we find that clones of neurones do not form radial columns; rather, they tend to occupy the same or neighbouring cortical laminae and to be spread over several hundreds of micrometers of cortex in the horizontal dimension. This spread occurs in both mediolateral and rostrocaudal directions.

scholarly journals Histamine-containing peripheral neuronal and endocrine systems.

1985 ◽  
Vol 33 (9) ◽  
pp. 933-941 ◽  
Author(s):  
P Panula ◽  
M Kaartinen ◽  
M Mäcklin ◽  
E Costa
Keyword(s):  
Mast Cells ◽  
Guinea Pig ◽  
Peripheral Nerves ◽  
Endocrine Cells ◽  
Ganglion Cell Layer ◽  
Cell Layer ◽  
Nerve Fibers ◽  
Immunohistochemical Method ◽  
Rat Stomach ◽  
Rat Ileum

An immunohistochemical method was developed to detect histamine in tissues. The aim of this study was to reveal the cellular stores of histamine in the gastrointestinal tract, pituitary, and adrenal gland. Histamine-containing nerve fibers were found in both rat and guinea pig gut. The origin of at least some of these fibers in the rat ileum was the submucous ganglion cell layer. In the rat stomach, numerous enterochromaffin-like cells exhibited histamine immunofluorescence, and endocrine cells in the ileum and jejunum contained histamine. Only mast cells contained histamine in the neurohypophysis. A large number of process-bearing cells in the guinea pig but not in the rat adrenal medulla contained histamine. The study shows that histamine is present in peripheral nerves and endocrine cells in addition to mast cells, and may function as a neurotransmitter or hormone.

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