Localization of PRAMEY Protein in Bovine Testis by Immunoelectron Microscopy

Gonadotroph cell adenomas of the pituitary are infrequent in human patients and are not invariably associated with altered gonadal function. To date, no animal model of this tumor type exists. Herein, we describe spontaneous gonadotroph cell adenomas in old male and female Sprague-Dawley rats by histology, immunocytology and electron microscopy.The material consisted of the pituitaries of 27 male and 38 female Sprague Dawley rats, all 26 months of age or older, removed at routine autopsy. Sections of formal in-fixed, paraffin-embedded tissue were stained with hematoxylin-phloxine-saffron (HPS), the PAS method and the Gordon-Sweet technique for the demonstration of reticulin fibers. For immunostaining, sections were exposed to anti-rat β-LH, anti-ratβ-TSH, anti-rat PRL, anti-rat GH and anti-rat ACTH 1-39. For electron microscopy, tissue was fixed in 2.5% glutaraldehyde, postfixed in 1% OsO4 and embedded in epoxy-resin. Tissue fixed in 10% formalin, embedded in epoxy resin without osmification, was used for immunoelectron microscopy.

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Solid-phase immunoelectron microscopy for rapid diagnosis of enteroviruses

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100111240 ◽

1984 ◽

Vol 42 ◽

pp. 226-227 ◽

Cited By ~ 1

Author(s):

K. Pegg-Feige ◽

F. W. Doane

Keyword(s):

Electron Microscopy ◽

Cell Culture ◽

Immunoelectron Microscopy ◽

Detection Method ◽

Solid Phase ◽

Protein A ◽

Plant Viruses ◽

Rapid Diagnosis ◽

Virus Diagnosis ◽

Antiviral Antibody

Immunoelectron microscopy (IEM) applied to rapid virus diagnosis offers a more sensitive detection method than direct electron microscopy (DEM), and can also be used to serotype viruses. One of several IEM techniques is that introduced by Derrick in 1972, in which antiviral antibody is attached to the support film of an EM specimen grid. Originally developed for plant viruses, it has recently been applied to several animal viruses, especially rotaviruses. We have investigated the use of this solid phase IEM technique (SPIEM) in detecting and identifying enteroviruses (in the form of crude cell culture isolates), and have compared it with a modified “SPIEM-SPA” method in which grids are coated with protein A from Staphylococcus aureus prior to exposure to antiserum.

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Detection of Viral Antigens in Mouse and Rat Tumor Cells by Immunoelectron Microscopy Following Periodate-Lysine-Paraformaldehyde (PLP) Fixation

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010009124x ◽

1976 ◽

Vol 34 ◽

pp. 252-253

Author(s):

Y. Ohtsuki ◽

G. Seman ◽

J. M. Bowen ◽

M. Scanlon ◽

L. Dmochowski

Keyword(s):

Immunoelectron Microscopy ◽

Mammary Tumor ◽

Type A ◽

Rabbit Serum ◽

Virus Particles ◽

Viral Antigens ◽

Culture Cells ◽

Mouse Mammary Tumor ◽

Tumor Virus ◽

Immunoperoxidase Labeling

Recently, periodate-lysine-paraformaldehyde (PLP) fixation was reported for immunoelectron microscopy (1). In PLP fixation, carbohydrates are oxidized by periodate and cross-linked by lysine; paraformaldehyde stabilizes proteins and lipids. By using PLP fixation, intracytoplasmic type A viral antigens have been previously demonstrated by immunoperoxidase labeling (2). In the present study, PLP fixation has been applied for the detection of the same antigens in mouse mammary tumor culture cells by both immunoferritin and immunoperoxidase methods. Rabbit anti-intracytoplasmic type A virus serum (anti-A), kindly provided by Dr. M. Muller (3), rabbit anti-strain A mouse mammary tumor virus (anti-MMTV) and preimmune rabbit serum as control were used to detect viral antigens in cells of C3H/HeJ strain mouse mammary tumor culture. Attempts have been also made to demonstrate peroxidase labeling of type C virus particles in frozen sections of an SD-MSV-induced NZB rat bone tumor tissue by rabbit anti-MuLV serum.

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Identification of maize mosaic virus by immunoelectron microscopy

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100142827 ◽

1986 ◽

Vol 44 ◽

pp. 240-241

Author(s):

O. E. Bradfute

Keyword(s):

Zea Mays ◽

Mosaic Virus ◽

Immunoelectron Microscopy ◽

Severe Disease ◽

Maize Mosaic Virus ◽

The World ◽

Plant Rhabdovirus

Maize mosaic virus (MMV) causes a severe disease of Zea mays in many tropical and subtropical regions of the world, including the southern U.S. (1-3). Fig. 1 shows internal cross striations of helical nucleoprotein and bounding membrane with surface projections typical of many plant rhabdovirus particles including MMV (3). Immunoelectron microscopy (IEM) was investigated as a method for identifying MMV. Antiserum to MMV was supplied by Ramon Lastra (Instituto Venezolano de Investigaciones Cientificas, Caracas, Venezuela).

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Banded aggregates containing fibrillin are present in the cartilage matrix adjacent to chondrocytes in individuals affected with scoliosis

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100124860 ◽

1992 ◽

Vol 50 (1) ◽

pp. 892-893

Author(s):

Douglas R. Keene ◽

Magaret Fairhurst ◽

Catherine C. Ridgway ◽

Lynn Y. Sakai

Keyword(s):

Connective Tissue ◽

Marfan Syndrome ◽

Cross Section ◽

Immunoelectron Microscopy ◽

Cartilage Matrix ◽

Negative Stain ◽

Gene Coding ◽

Rotary Shadowing

Matrix microfibrils are present in the connective tissue matrices of all tissues. Following standard TEM processing, they appear in cross section as cylindrical fibrils 8-10 nm in diameter, often associated with amorphous elastin. They are also seen in the absence of amorphous elastin, for example in the shallow papillary layer of skin, and also in cartilage matrix (Figure 1). Negative stain and rotary shadowing studies suggest that microfibrils are composed of laterally associated globular structures connected by fine filamentous strands (“ beaded strings”), and that they are extendable. Immunoelectron microscopy has demonstrated that fibrillin, a 350 Kd glycoprotein, is distributed along all microfibrils with a relaxed periodicity of about 54 nm The gene coding for fibrillin has recently been identified and is defective in the Marfan syndrome.

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Immunocytochemical studies on the behavior of RuBisCO and LHCP II during the cell cycle of synchronized Euglena gracilis

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100160868 ◽

1990 ◽

Vol 48 (3) ◽

pp. 662-663

Author(s):

Tetsuaki Osafune ◽

Shuji Sumida ◽

Tomoko Ehara ◽

Eiji Hase ◽

Jerome A. Schiff

Keyword(s):

Cell Cycle ◽

Immunoelectron Microscopy ◽

Growth Phase ◽

Euglena Gracilis ◽

Dark Period ◽

Light Period ◽

Carboxylase Activity ◽

Immunocytochemical Studies ◽

Lhcp Ii

Changes in the morphology of pyrenoid and the distribution of RuBisCO in the chloroplast of Euglena gracilis were followed by immunoelectron microscopy during the cell cycle in a light (14 h)- dark (10 h) synchronized culture under photoautotrophic conditions. The imrnunoreactive proteins wereconcentrated in the pyrenoid, and less densely distributed in the stroma during the light period (growth phase, Fig. 1-2), but the pyrenoid disappeared during the dark period (division phase), and RuBisCO was dispersed throughout the stroma. Toward the end of the division phase, the pyrenoid began to form in the center of the stroma, and RuBisCO is again concentrated in that pyrenoid region. From a comparison of photosynthetic CO2-fixation with the total carboxylase activity of RuBisCO extracted from Euglena cells in the growth phase, it is suggested that the carboxylase in the pyrenoid functions in CO2-fixation in photosynthesis.

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Examination of cutaneous macroglobulinosis by immunoelectron microscopy

British Journal of Dermatology ◽

10.1046/j.1365-2133.1996.d01-990.x ◽

1996 ◽

Vol 135 (2) ◽

pp. 287-291 ◽

Cited By ~ 1

Author(s):

D. LIPSKER ◽

B. CRIBIER ◽

D. SPEHNER ◽

N. BOEHM ◽

E. HEID ◽

...

Keyword(s):

Immunoelectron Microscopy

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Internalization of an Anti-Glycoprotein IIb/IIIa Antibody by HEL Cells

Thrombosis and Haemostasis ◽

10.1055/s-0038-1653767 ◽

1995 ◽

Vol 73 (02) ◽

pp. 291-296 ◽

Cited By ~ 1

Author(s):

Kenjiro Hamamoto ◽

Shosaku Nomura ◽

Masahiko Suzuki ◽

Shigetoshi Ohga ◽

Shirou Fukuhara

Keyword(s):

Flow Cytometry ◽

Monoclonal Antibodies ◽

Immunoelectron Microscopy ◽

Surface Membrane ◽

Intracellular Pool ◽

Adhesion Proteins ◽

Hel Cells

SummaryPlatelets are known to internalize monoclonal antibodies directed against the glycoprotein (GP) IIb/IIIa complex. We investigated whether an antibody directed against this complex (NNKY 2-11) was transported from the surface membrane to the intracellular pool in HEL cells. Flow cytometry showed that the percent binding of NNKY 2-11 to the surface membrane of HEL cells was decreased after incubation for 24 h compared with 1 h, while the binding of an anti-GPIb antibody (NNKY 5-5) did not change. It did not seem likely that the GP Ilb/IIIa complex antibody was shed from the surface membrane of the HEL cells during incubation, because the medium conditioned by incubation with these cells for 24 h showed almost no binding to washed platelets. In addition, immunoelectron microscopy demonstrated that GP IIb/IIIa complex antibodies were incorporated into the intracellular pool of HEL cells and were associated with alpha granules. These findings indicated that an anti-GP IIb/IIIa antibody could be internalized by megakaryocytes, as has been previously shown with platelets, suggesting that megakaryocyte GP IIb/IIIa may act as a carrier for various adhesion proteins.

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