Immunogold localization of HMB-45 antigen in pulmonary lymphangiomyomatosis

1995 ◽  
Vol 53 ◽  
pp. 998-999
Author(s):  
J.M. Minda ◽  
E. Dessy ◽  
G. G. Pietra
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Room Temperature ◽  
Osmium Tetroxide ◽  
Muscle Cells ◽  
Ultrastructural Localization ◽  
Reproductive Age ◽  
Thin Sections ◽  
Smooth Muscle Proliferation ◽  
Hmb 45

Pulmonary lymphangiomyomatosis (PLAM) is a rare disease occurring exclusively in women of reproductive age. It involves the lungs, lymph nodes and lymphatic ducts. In the lungs, it is characterized by the proliferation of smooth muscle cells around lymphatics in the bronchovascular bundles, lobular septa and pleura The nature of smooth muscle proliferation in PLAM is still unclear. Recently, reactivity of the smooth muscle cells for HMB-45, a melanoma-related antigen has been reported by immunohistochemistry. The purpose of this study was the ultrastructural localization of HMB-45 immunoreactivity in these cells using gold-labeled antibodies.Lung tissue from three cases of PLAM, referred to our Institution for lung transplantation, was embedded in either Poly/Bed 812 post-fixed in 1% osmium tetroxide, or in LR White, without osmication. For the immunogold technique, thin sections were placed on Nickel grids and incubated with affinity purified, monoclonal anti-melanoma antibody HMB-45 (1:1) (Enzo Diag. Co) overnight at 4°C. After extensive washing with PBS, grids were treated with Goat-anti-mouse-IgG-Gold (5nm) (1:10) (Amersham Life Sci) for 1 hour, at room temperature.

scholarly journals Extrarenal angiomyolipoma in uterine cervix: rare presentation in unusual site

2019 ◽  
Vol 8 (1) ◽  
pp. 367
Author(s):  
Rashmi Monteiro ◽  
Shikha Sharma ◽  
Sonal Gupta ◽  
Indu Choudhary
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Blood Vessels ◽  
Uterine Cervix ◽  
Abdominal Mass ◽  
Muscle Cells ◽  
Renal Angiomyolipoma ◽  
Rare Presentation ◽  
Unusual Site ◽  
Hmb 45

Angiomyolipoma is a benign neoplasm composed of variable admixture of blood vessels, smooth muscle cells and adipose tissue. Cervical angiomyolipoma are extremely rare and to the best of our knowledge only five cases of angiomyolipoma in cervix have been reported in the literature till date. Authors are presenting a case of angiomyolipoma arising from the uterine cervix. 43 years old female presented with mass descending per vagina for 6 months. This case had no association with tuberous sclerosis. Microscopic examination showed an ill-defined polypoidal, non-encapsulated lesion covered by keratinized stratified epithelium. The lesion is made up of three components, predominantly by fascicles of spindle shaped cells, varying sized blood vessels and multiple foci of mature adipocytes with no evidence of atypia or increased mitotic activity. Smooth muscle component showed strong immunoreactivity to SMA and absence of elastic fibres in the blood vessels were confirmed by histochemistry. Non-vascular smooth muscle cells were negative for HMB-45 in contrast to renal and other extra-renal angiomyolipoma in which HMB-45 immunoreactivity in seen in these cells. To conclude, the differential diagnosis of lower abdominal mass and dysfunctional uterine bleeding should include the angiomyolipoma, even though the uterine cervix is an extremely rare location where they occur.

scholarly journals ULTRASTRUCTURAL INTERRELATIONSHIPS OF NERVE PROCESSES AND SMOOTH MUSCLE CELLS IN THREE DIMENSIONS

1966 ◽  
Vol 28 (1) ◽  
pp. 37-49 ◽  
Author(s):  
J. C. Thaemert
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Osmium Tetroxide ◽  
Three Dimensional ◽  
Muscle Cells ◽  
Uranyl Acetate ◽  
Intestinal Wall ◽  
Three Dimensions ◽  
Serial Sections ◽  
Muscularis Externa

The muscularis externa of the intestinal wall of frogs was fixed in osmium tetroxide, embedded in Vestopal-W, serially sectioned for electron microscopy, and stained with uranyl acetate. A method to obtain individually mounted and properly positioned serial sections is described. The three-dimensional techniques used during the course of this investigation demonstrate that it is possible to examine carefully relatively large areas of tissue on individual serial sections with the electron microscope and subsequently to construct montages of electron micrographs of pertinent areas from each section. Several carefully rendered interrelationships of nerve processes and smooth muscle cells in three dimensions are exhibited and described. Recent studies of other neuro-effector relationships are discussed in relation to the present status of the nature and organization of the autonomic nervous system in visceral organs.

scholarly journals Proteoglycans in primate arteries. I. Ultrastructural localization and distribution in the intima.

1975 ◽  
Vol 67 (3) ◽  
pp. 660-674 ◽  
Author(s):  
T N Wight ◽  
R Ross
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Muscle Cells ◽  
Ultrastructural Localization ◽  
Ruthenium Red ◽  
Basement Membranes ◽  
Macaca Nemestrina ◽  
Elastic Fibers ◽  
Surface Coat ◽  
The Matrix

Proteoglycans were identified and localized histochemically and ultrastructurally in normal and hyperplastic arterial intimas in nonhuman primates (Macaca nemestrina). These regions were consistently more alcianophilic than the adjacent medial layers and this alcianophilia was absent after treatment with glycosaminoglycan-degradative enzymes. Ultrastructurally, the intimal intercellular matrix consisted of numerous, irregularly shaped, 200-500-A diameter granules possessing 30--60-A diameter filamentous projections, and these granules were dispersed between collagen and elastic fibers. The granules exhibited a marked affinity for ruthenium red and were interconnected via their filamentous projections. The ruthenium red-positive granules were intimately associated with the plasma membrane of intimal smooth muscle cells and attached to collagen fibrils and elastic fibers. The matrix granules were completely removed after testicular hyaluronidase or chondroitinase ABC digestion but only partially removed after leech hyaluronidase treatment. These results suggest that the matrix granules contain some hyaluronic acid and one or more isomers of chondroitin sulfate. In addition to the large ruthenium red-positive matrix granules, a smaller class of ruthenium red-positive granule (100--200-A diameter) was present within the basement membranes beneath the endothelium and surrounding the smooth muscle cells. Ruthenium red also exhibited an affinity for the surface coat of the smooth muscle cells. The potential importance of proteoglycans in arterial intimal hyperplasia is discussed.

scholarly journals An electron stain for elastic fibers using orcein.

1977 ◽  
Vol 25 (4) ◽  
pp. 306-308 ◽  
Author(s):  
H Nakamura ◽  
C Kanai ◽  
V Mizuhira
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Muscle Cells ◽  
Uranyl Acetate ◽  
Elastic Fibers ◽  
Lead Citrate ◽  
Thin Sections ◽  
Electron Microscopes ◽  
Orcein Staining

Orcein was found to be useful as an electron-opaque stain for elastic fibers in epoxy-sections. Ultra-thin sections of aorta were treated with elastica stain containing 0.1-0.3% orcein and counterstained in uranyl acetate and lead citrate. Elastic fibers were densely and specifically demonstrated in the stroma and near smooth muscle cells. The result of orcein staining has a comparable appearance under both light and electron microscopes.

scholarly journals A STUDY OF NEXUSES IN VISCERAL SMOOTH MUSCLE

1969 ◽  
Vol 41 (1) ◽  
pp. 287-297 ◽  
Author(s):  
J. L. S. Cobb ◽  
Terence Bennett
Keyword(s):  
Smooth Muscle ◽  
Guinea Pig ◽  
Smooth Muscle Cells ◽  
Osmium Tetroxide ◽  
Vas Deferens ◽  
Muscle Cells ◽  
Taenia Coli ◽  
Muscle Tissues ◽  
Fixation Techniques ◽  
Visceral Smooth Muscle

Nexuses are described between the smooth muscle cells of the gizzard of the chick and the pigeon, the vas deferens of the mouse and the guinea pig, and the taenia coli of the guinea pig. The nexuses in the gizzard were examined after osmium tetroxide and potassium permanganate had been used as fixatives. Although differences in the dimensions of the unit membranes and the nexuses were noted, the results with the two fixation techniques were complementary. The distribution of nexuses within the smooth muscle tissues examined was uneven. Nexuses were still present in both small and large pieces of tissue incubated in hypertonic solutions at varying temperatures. Other experiments showed that the degree of contraction at the time of fixation did not affect the presence of nexuses in the tissue. These results indicate that nexuses between smooth muscle cells are stable under a variety of conditions.

scholarly journals Immunoelectron microscope studies of membrane-microfilament interactions: distributions of alpha-actinin, tropomyosin, and vinculin in intestinal epithelial brush border and chicken gizzard smooth muscle cells.

1981 ◽  
Vol 91 (3) ◽  
pp. 614-628 ◽  
Author(s):  
B Geiger ◽  
A H Dutton ◽  
K T Tokuyasu ◽  
S J Singer
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Brush Border ◽  
Muscle Cells ◽  
Ultrastructural Localization ◽  
Cytoskeletal Proteins ◽  
Junctional Complex ◽  
Intestinal Epithelial ◽  
Chicken Gizzard ◽  
Dense Bodies

The ultrastructural localization of three cytoskeletal proteins, alpha-actinin, tropomyosin, and vinculin, in the brush border of epithelial cells of chicken small intestine and the smooth muscle cells of chicken gizzard was studied by immunofluorescence and immunonelectron microscope labeling of frozen sections of lightly fixed, intact tissues. In the immunoelectron microscope studies, a recently described new type of electron-dense antibody conjugate, imposil-antibody, has been successfully used, along with ferritin-antibody conjugates, in single and double immunolabeling experiments. In the intestinal brush border shows that vinvulin is sharply confined to the junctional complex close to the membrane region of the zonula adherens, in distinct contrast to the more diffuse distributions of the other two proteins. In the smooth muscle cells, the labeling patterns show that vinculin is sharply confined to the membrane-associated dense plaques, closer to the membrane than the alpha-Actinin is also present in the cytoplastic dense bodies, from which vinculin is absent. Tropomyosin is present diffusely distributed in the cytoplasm, but absent from both dense plaques and dense bodies. These findings with the muscle cells demonstrate, therefore, that the dense plaques and dense bodies are chemically and structurally distinct entities. The results with both tissues, along with those in previous papers (Geiger, 1979, Cell. 18:193-205.; Geiger et al., 1980, Proc. Natl. Acad. Sci. U. S. A. 77:4127-4131), suggest that vinculin may play an important and widespread role in the linkage of actin-containing microfilament bundles to membranes.

scholarly journals MYOSIN-LIKE AGGREGATES IN TRYPSIN-TREATED SMOOTH MUSCLE CELLS

1971 ◽  
Vol 48 (1) ◽  
pp. 174-188 ◽  
Author(s):  
Jack Rosenbluth
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Muscle Protein ◽  
Muscle Cells ◽  
Smooth Muscle Myosin ◽  
Stable Complex ◽  
Solution Ph ◽  
Thin Sections ◽  
Thick Filaments ◽  
Muscle Myosin

Segments of the lower small intestine of the toad Bufo marinus were excised and soaked for approximately 2 hr in Ringer's solution (pH 7.4 or 7.8) containing crystalline trypsin and then fixed for electron microscopy at approximately the same pH. Thin sections of the tunica muscularis of these specimens show smooth muscle cells ranging in appearance from severely damaged at one extreme to apparently unaffected at the other. Among these are cells at intermediate stages, including some which exhibit large and conspicuous populations of thick filaments closely resembling artificially prepared aggregates of smooth muscle myosin. The thick filaments have the form of tactoids ∼ 250–300 A in diameter in their middle regions and are ∼ 0.5–1.0 µ in length. In some preparations they also display an axial periodicity approximating 143 A. They are usually randomly oriented and segregated from the thin filaments, which tend to form closely packed, virtually crystalline bundles at the periphery of these cells. "Dense bodies" are absent from cells showing these changes. The simplest interpretation of these data is that smooth muscle myosin normally exists among the actin filaments in a relatively disaggregated state and that trypsin induces aggregation by altering the conformation of the myosin molecule. Alternatively, trypsin may act indirectly through an effect on some other smooth muscle protein which normally forms a stable complex with relatively disaggregated myosin.

scholarly journals Involvement of extracellular-matrix-degrading metalloproteinases in rabbit aortic smooth-muscle cell proliferation

1992 ◽  
Vol 288 (1) ◽  
pp. 93-99 ◽  
Author(s):  
K M Southgate ◽  
M Davies ◽  
R F G Booth ◽  
A C Newby
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Muscle Cells ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Aortic Smooth Muscle ◽  
Affect Cell Viability ◽  
Smooth Muscle Proliferation ◽  
Aortic Explants ◽  
Interstitial Collagenase

We investigated the influence of two structurally unrelated inhibitors of matrix-degrading metalloproteinases, Ro 31-4724 and Ro 31-7467, on the primary proliferation of smooth-muscle cells from rabbit aortic explants. Both agents inhibited proliferation in a concentration-dependent manner, but did not affect cell viability. Smooth-muscle cells grown out from explants secreted 95 kDa and 72 kDa gelatinase enzymes that were also inhibited in a concentration-dependent manner by Ro 31-4724 and Ro 31-7467. Interstitial collagenase and stromelysin were not detected. We conclude that metalloproteinases are likely to be involved in the initiation of smooth-muscle proliferation.

Ultrastructure of myoepithelial cell in parotid gland

1988 ◽  
Vol 46 ◽  
pp. 342-343
Author(s):  
C. N. Sun
Keyword(s):  
Parotid Gland ◽  
Osmium Tetroxide ◽  
Individual Cell ◽  
Branching Processes ◽  
Muscle Cells ◽  
Myoepithelial Cells ◽  
Uranyl Acetate ◽  
Thin Sections ◽  
Gland Carcinoma ◽  
Parotid Gland Carcinoma

Myoepithelial cells have been observed in the prostate, harderian, apocrine, exocrine sweat and mammary glands. Such cells and their numerous branching processes form basket-like structures around the glandular acini. Their shapes are quite different from structures seen either in spindleshaped smooth muscle cells or skeletal muscle cells. These myoepithelial cells lie on the epithelial side of the basement membrane in the glands. This presentation describes the ultrastructure of such myoepithelial cells which have been found also in the parotid gland carcinoma from a 45-year old patient.Specimens were cut into small pieces about 1 mm3 and immediately fixed in 4 percent glutaraldehyde in phosphate buffer for two hours, then post-fixed in 1 percent buffered osmium tetroxide for 1 hour. After dehydration, tissues were embedded in Epon 812. Thin sections were stained with uranyl acetate and lead citrate. Ultrastructurally, the pattern of each individual cell showed wide variations.

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