scholarly journals Immunohistochemical localization of acetylcholine receptors at human endplates using a monoclonal antibody.

1987 ◽  
Vol 35 (5) ◽  
pp. 613-617 ◽  
Author(s):  
L M Smit ◽  
H Veldman ◽  
F G Jennekens
Keyword(s):  
Monoclonal Antibody ◽  
Acetylcholine Receptors ◽  
Immunohistochemical Localization ◽  
Immunohistochemical Method ◽  
Electron Microscopic Level ◽  
Immunoperoxidase Technique ◽  
Electron Microscopic ◽  
Fixed Tissue ◽  
Alpha Bungarotoxin ◽  
Diagnostic Investigations

We describe a simple indirect immunohistochemical method for localization of acetylcholine receptors (AChR) in motor endplates at the light and electron microscopic level. This method involves the use of a monoclonal antibody directed against the main immunogenic region (MIR) of AChRs and is applicable to periodate-lysine-paraformaldehyde (PLP)-fixed tissue. We discuss the advantages of this method, as compared with the alpha-bungarotoxin-immunoperoxidase technique, and stress its value for diagnostic investigations of motor point biopsies from patients with neuromuscular transmission disorders.

scholarly journals Influence of neostigmine treatment on embryonic development of acetylcholine receptors and neuromuscular junctions.

1982 ◽  
Vol 94 (3) ◽  
pp. 540-548 ◽  
Author(s):  
G S Sohal ◽  
W R Boydston
Keyword(s):  
Embryonic Development ◽  
Acetylcholine Receptor ◽  
Acetylcholine Receptors ◽  
Neuromuscular Junctions ◽  
Electron Microscopic Level ◽  
Oblique Muscle ◽  
Apparent Effect ◽  
Alpha Bungarotoxin ◽  
Superior Oblique Muscle ◽  
Superior Oblique

The postulated role of the acetylcholine receptor in the formation of neuromuscular synapses during the course of embryonic development was investigated in the superior oblique muscle of white Peking duck embryos. The possibility that the number of receptors could be experimentally lowered by chronic injections of the anticholinesterase agent, neostigmine methylsulfate, was determined using 125I-alpha-bungarotoxin. The total number of acetylcholine receptors on incubation day 12, 2 d subsequent to the onset of treatment, was reducted 45% as compared to saline-treated controls. A similar reduction in total receptor content (49%) was also observed on day 19. Radioautographic preparations showed that clusters of acetylcholine receptors were rare and that the grain density of extrajunctional receptors was also reduced. Hence, chronic treatment with neostigimine during development was observed to exert an effect on both the number and distribution of receptors in the developing superior oblique muscle. These changes occurred in the absence of any apparent effect on muscle differentiation in general. Myoblasts and myotubes were present on day 14 and further differentiated into myofibers by day 18 in both neostigmine and saline-treated muscles. The cytology of the develop;ing muscle cells also appeared normal. This is in contradistinction to the striking morphological changes that take place in adult mammalian and avian muscle after anticholinesterase treatment. More significantly, the decreased total receptor content and sparsity of clusters had no apparent effect on the formation of developing neuromuscular junctions at the electron microscopic level. The frequency of neuromuscular junctions in neostigmine-treated muscles was similar to that of the controls. It is concluded that acetylcholine receptor clusters are not required for the events leading to the morphological formation of neuromuscular junctions during in vivo development.

scholarly journals Application of avidin-biotin-peroxidase complex method with a monoclonal antibody to human Ia-like antigen in immunoelectron microscopy.

1986 ◽  
Vol 34 (11) ◽  
pp. 1495-1499 ◽  
Author(s):  
K C Feng-Chen ◽  
B F Chen ◽  
Z Liu ◽  
A K Ng
Keyword(s):  
Monoclonal Antibody ◽  
Ultrastructural Level ◽  
Electron Microscopic Level ◽  
Complex Method ◽  
Outer Cell ◽  
Electron Microscopic ◽  
Hla Dr ◽  
Combined Use ◽  
Antibody Complex ◽  
Reticular Cells

Monoclonal antibody (MAb) to human Ia-like (HLA-DR) antigen was applied with the avidin-biotin-peroxidase complex (ABC) immunostaining method to localize the Ia-like antigen at the electron microscopic level. Our results indicated that in human tonsils and adenoids fixed with 4-6% phosphate-buffered paraformaldehyde for 4-6 hr, sharply delineated electron-dense products of the antigen and antibody complex were detectable on the outer cell membranes of lymphoblasts, lymphocytes, reticular cells, and macrophages. In our study, the vibratome sections of the paraformaldehyde-fixed, pre-embedding immunostained tissues consistently showed more satisfactory morphology than frozen sections. The combined use of the anti-human Ia monoclonal antibody and the ABC procedure with paraformaldehyde fixation provides a simple and sensitive method to study at the ultrastructural level the Ia-like antigen-bearing cells, which are vital in the immune response.

scholarly journals PREGNANCY, COMPLICATED BY PREECLAMPSIA: FETOPLACENTAL COMPLEX IMMUNE DEADAPTATION AND HISTOSTRUCTURAL FEATURES

2020 ◽  
Vol 73 (1) ◽  
pp. 99-103
Author(s):  
Pavlo V. Yavorskyi ◽  
Vitalii M. Zozulia ◽  
Oleh Ya. Vanchuliak ◽  
Marta S. Garazdiuk
Keyword(s):  
Pregnant Women ◽  
Transforming Growth Factor ◽  
Morphological Changes ◽  
Immunohistochemical Method ◽  
Electron Microscopic Level ◽  
Control Group ◽  
Electron Microscopic ◽  
Correlation Relationship ◽  
Uncomplicated Pregnancy ◽  
Fetoplacental Complex

The aim: to study and compare the features of the interleukins levels and morphological changes of placenta at various stages of preeclampsia. Materials and methods: 109 pregnant women with preeclampsia of varying severity (study group) and 30 pregnant women with uncomplicated pregnancy (control group) were examined. Immunohistochemical method, proinflammatory interleukins levels, morphological and morphometric analysis of peripheral and central placental areas biopsies on the optical and electron-microscopic level have been used. Results: Morphofunctional changes in the placenta in case of preeclampsia and the increase in the expression level of the transforming growth factor have a series of regular stages from the formation, strain and disruption of adaptive mechanisms with more pronounced signs of morphological immaturity of parenchymal and stromal elements of the placenta, especially in the area of syncytiotrophoblast and spiral vessels. The degree of clinical manifestation of preeclampsia has a correlation relationship with IL-10 deficiency and with the increase in TNF-α, stimulation of macrophage-protein production that contributes to the change in the ratio of Thl / Th2, which are antagonists and inhibit each other’s development. Conclusions: The severity of the preeclampsia course correlates with the state of morphofunctional changes in the placenta and changes in the ratio of the pro- and anti-inflammatory interleukins.

Monoclonal antibody study of the decorated spongiome of contractile vacuole complexes of Paramecium

1990 ◽  
Vol 96 (3) ◽  
pp. 469-475
Author(s):  
R.D. Allen ◽  
M.S. Ueno ◽  
L.W. Pollard ◽  
A.K. Fok
Keyword(s):  
Monoclonal Antibody ◽  
Developmental Stages ◽  
General Scheme ◽  
Central Zone ◽  
Immunogold Labeling ◽  
Microscopic Level ◽  
Contractile Vacuole ◽  
Electron Microscopic Level ◽  
Frozen Sections ◽  
Electron Microscopic

A monoclonal antibody (mAb) has been developed and selected by immunofluorescence for the radial canals of the contractile vacuole complex (CVC) of Paramecium multimicronucleatum. By applying indirect immunogold labeling to thin frozen sections this mAb has been shown at the electron microscopic level to be specific for the decorated spongiome. We have used the mAb to study the normal interfission appearance as well as developmental stages of the decorated spongiomes. Two decorated spongiomes, presumably involved in water sequestration, radiate as 5–10 bands from unlabeled, circular, 25 microns diameter centers. Two new CVCs arise just anterior to the space occupied by the old spongiomes, the new anterior CVC appearing slightly before the posterior one. Development of the new spongiomes around a 10 microns unlabeled central zone is accompanied by a regression of old spongiome bands until the lengths of these bands in both old and new CVCs are equal just before cell division. After division both old and new spongiome bands grow at equal rates to the same length. Exceptions to the above general scheme, both in number of CVCs in interfission, as well as in position of the new relative to the old CVCs, are also observed.

scholarly journals Efficient immunodetection of various protein antigens in glutaraldehyde-fixed brain tissue.

1995 ◽  
Vol 43 (12) ◽  
pp. 1285-1291 ◽  
Author(s):  
A Mrini ◽  
H Moukhles ◽  
H Jacomy ◽  
O Bosler ◽  
G Doucet
Keyword(s):  
Brain Tissue ◽  
Polyclonal Antibodies ◽  
Electron Microscopic Level ◽  
Lower Sensitivity ◽  
Glutaraldehyde Fixation ◽  
Protein Antigens ◽  
Neurofilament Protein ◽  
Electron Microscopic ◽  
Fixed Tissue ◽  
Methyl Transferase

Optimal ultrastructural preservation of brain tissue for electron microscopy is best achieved with fixatives containing high concentrations of glutaraldehyde, which is generally considered detrimental to the immunogenicity of most protein antigens. We tested seventeen mono- or polyclonal antibodies against peptide or protein antigens, including a majority for which immunoreactivity had previously been reported to be sensitive to glutaraldehyde fixation. Forebrain sections of rats or mice fixed by perfusion with 3.5% glutaraldehyde were processed for pre-embedding immunocytochemistry by the avidin-biotin method. The resulting immunostaining was in most cases at least similar to that obtained in sections fixed with paraformaldehyde. Immunoreactivity against the mouse or human neurofilament protein NF-L was even improved, being similar to that previously reported for unfixed brain tissue. Of all antigens tested, only choline acetyltransferase, phenylethanolamine-N-methyl transferase, and neuropeptide Y were detected with lower sensitivity than after paraformaldehyde fixation, which was attributed to a rather restricted penetration of the primary antibody into glutaraldehyde-fixed tissue sections. These results indicate that glutaraldehyde may be envisaged as a possible fixative for optimal immunocytochemical detection of any tissue antigen at the electron microscopic level, including antigens which, on the basis of results obtained after fixation with paraformaldehyde-glutaraldehyde mixtures, were considered highly sensitive to glutaraldehyde fixation.

scholarly journals Distribution of the slow/cardiac isoform of skeletal muscle Ca(2+)-ATPase in developing and mature tissues of chickens determined using a monoclonal antibody.

1993 ◽  
Vol 41 (2) ◽  
pp. 215-224
Author(s):  
S Shahin ◽  
P F Bartlett ◽  
T J Millar ◽  
I McLennan ◽  
J A Rostas
Keyword(s):  
Skeletal Muscle ◽  
Monoclonal Antibody ◽  
Smooth Muscle ◽  
Muscle Fibers ◽  
Cell Types ◽  
Electron Microscopic Level ◽  
Electron Microscopic ◽  
Adult Chicken ◽  
Intrafusal Muscle Fibers ◽  
Anterior Latissimus Dorsi

We have established that the monoclonal antibody (MAb) AA21, raised against a crude sarcolemmal fraction prepared from adult chicken anterior latissimus dorsi muscle, recognizes the slow twitch/cardiac isoform of calcium ATPase. This was done using a combination of immunohistochemistry at the light and electron microscopic level, the change in the cell distribution in skeletal muscle during development, the molecular weight of the principal protein recognized in Western transfers, and direct comparison with another MAb of known specificity. The antigen is initially expressed by all myotubes at E10 and with development is gradually lost from all presumptive fast fibers. In addition to its immunoreaction and slow extrafusal skeletal muscle fibers, AA21 displays a highly selective immunoreactivity with a number of other cell types in different tissues. The antibody stains a subset of intrafusal muscle fibers and intestinal and arterial smooth muscle, but not venous smooth muscle. In the nervous system, a subpopulation of neurons is intensely stained, most neurons are faintly stained, and glia are not stained at all.

scholarly journals Intracellular localization of thymosin alpha 1 by immunoelectron microscopy using a monoclonal antibody.

1987 ◽  
Vol 35 (2) ◽  
pp. 181-187 ◽  
Author(s):  
C Auger ◽  
C Stahli ◽  
N Fabien ◽  
J C Monier
Keyword(s):  
Monoclonal Antibody ◽  
Epithelial Cells ◽  
Intracellular Localization ◽  
Immunofluorescence Assay ◽  
Secretory Process ◽  
Electron Microscopic Level ◽  
Electron Microscopic ◽  
Thymic Hormone ◽  
Thin Sections ◽  
Thymosin Alpha 1

Distribution of thymosin alpha 1 in normal mice (OF1) or autoimmune mice (NZB) was investigated using immunocytochemical techniques on sections of GMA- and Epon-embedded mouse thymuses. A monoclonal antibody directed against synthetic thymosin alpha 1 was used. With the immunofluorescence assay, patchy staining of thymosin alpha 1 was found in the cytoplasm of epithelial cells of the subcapsullary and medullary zones of OF1 thymus. In NZB thymus, the fluorescent pattern was less precisely localized. At the electron microscopic level, immunolabeling of Epon-embedded ultra-thin sections revealed ferritin in some vacuoles of epithelial cells. Ferritin labeling in OF1 thymus was found in several small vacuoles of the same cell, but was present in large, dense vacuoles in NZB thymus. These differences might reflect differences in the secretory process of thymic hormone.

scholarly journals Ultrastructural immunohistochemical localization of adrenocorticotropin and beta-lipotropin in the rat brain.

1979 ◽  
Vol 27 (6) ◽  
pp. 1046-1048 ◽  
Author(s):  
G Pelletier
Keyword(s):  
Rat Brain ◽  
Neuronal Cell ◽  
Immunohistochemical Localization ◽  
Microscopic Level ◽  
Dense Core ◽  
Electron Microscopic Level ◽  
Serial Sections ◽  
Electron Microscopic ◽  
Dense Core Vesicles ◽  
Neuronal Cell Bodies

In an attempt to identify the cells and organellel containing ACTH and beta-lipotropin in the rat brain, an immunocytochemical localization of these two peptides was performed at the electron microscopic level. Both ACTH and beta-lipotropin were localized in dense core vesicles of about 60-80 nm in diameter. Using serial sections, it has been possible to demonstrate that these peptides are contained not only in the same neuronal cell bodies, but also in the same dense core vesicles.

scholarly journals A new high molecular mass protein showing unique localization in desmosomal plaque.

1989 ◽  
Vol 109 (4) ◽  
pp. 1511-1518 ◽  
Author(s):  
Y Hieda ◽  
S Tsukita ◽  
S Tsukita
Keyword(s):  
Monoclonal Antibody ◽  
Molecular Mass ◽  
High Molecular Mass ◽  
Microscopic Level ◽  
Electron Microscopic Level ◽  
Electron Microscopic ◽  
Binding Ability ◽  
Stratified Epithelium ◽  
Keratin Filaments

A high molecular mass protein of 680 kD was identified and purified from the isolated desmosomes in bovine muzzle epidermal cells. This protein, called "desmoyokin" (from the English, yoke) here, showed no binding ability with keratin filaments in vitro, and its molecule had a characteristic dumbell shape approximately 170 nm in length. We have succeeded in obtaining one monoclonal antibody specific to desmoyokin. By the use of this monoclonal antibody and antidesmoplakin monoclonal antibody, desmoyokin was shown to be colocalized with desmoplakin at the immunofluorescence microscopic level; desmoyokin occurred only in the stratified epithelium, not in the simple epithelium nor in the other tissues. At the electron microscopic level, these two proteins were clearly seen to be sorted out in the plaque of desmosomes with desmoyokin at the periphery and desmoplakin at the center of the disk-shaped desmosomal plaque, suggesting that these two plaque proteins play distinct roles in forming and maintaining the desmosomes in stratified epithelium.

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