Ultrastructural localization of actin in muscle, epithelial and secretory cells by applying the protein A-gold immunocytochemical technique

1983 ◽  
Vol 15 (1) ◽  
pp. 39-58 ◽  
Author(s):  
Mo�se Bendayan
Keyword(s):  
Protein A ◽  
Ultrastructural Localization ◽  
Secretory Cells ◽  
Immunocytochemical Technique

Ultrastructural localization of cytoskeletal proteins in pancreatic secretory cells

1985 ◽  
Vol 63 (6) ◽  
pp. 680-690 ◽  
Author(s):  
Moïse Bendayan
Keyword(s):  
Secretory Granules ◽  
Protein A ◽  
Ultrastructural Localization ◽  
Cytoskeletal Proteins ◽  
Secretory Cells ◽  
Secretory Proteins ◽  
Protein Maturation ◽  
Pancreatic Cells ◽  
Pancreatic Exocrine ◽  
Cell Web

Actin, myosin, and keratin immunoreactive sites have been localized with high resolution in pancreatic exocrine cells, by applying the protein A – gold technique on tissues processed at low temperature conditions. The labeling by gold particles was found at the level of the cell web and closely associated with the limiting membranes of the immature and mature secretory granules, as well as those of the "trans" cisternae of the Golgi apparatus. These results, together with those obtained in the study on the localization of secretory proteins in exocrine pancreatic cells, demonstrate that cytoskeletal proteins are present at sites where maturation and (or) concentration of the secretory proteins occur. Thus, besides the role that cytoskeletal proteins must play in the transport of the secretory granules from the Golgi to the plasma membrane, they may also be involved in the process of protein maturation and (or) concentration.

Isolation and ultrastructural localization of a soluble protein from Ophiostoma ulmi

1990 ◽  
Vol 68 (11) ◽  
pp. 2517-2524 ◽  
Author(s):  
R. S. Jeng ◽  
A. M. Svircev
Keyword(s):  
Polyclonal Antibodies ◽  
Polyacrylamide Gel Electrophoresis ◽  
Protein A ◽  
Leading Edge ◽  
Ultrastructural Localization ◽  
Immunogold Labelling ◽  
Two Dimensional ◽  
Thin Sections ◽  
Ophiostoma Ulmi ◽  
Gold Label

Two-dimensional polyacrylamide gel electrophoresis was used to identify and isolate a soluble polypeptide, the QP1 protein, which is characteristic of the vegetative hyphae of nonaggressive isolate Q412 of Ophiostoma ulmi. Individual QP1 spots were excised from 16 two-dimensional gels. Polypeptides were eluted from the gel spots by electroelution and lyophilized. The protein was injected into rabbits for the production of polyclonal antibodies. Antiserum specificity was tested by transferring polypeptides from a two-dimensional gel onto nitrocellulose and treating with QP1 serum. The resulting immunoblot contained a single spot that corresponded in shape and location to that of the QP1 polypeptide. Thin sections of fungal mycelia, from nonaggressive isolate Q412 and the aggressive isolate VA of O. ulmi, were treated with QP1 antibodies and protein A – gold. The gold label was localized in thin sections over conidial and hyphal cell walls of the nonaggressive isolate. The aggressive isolate was nonreactive. Mycelia from nonaggressive isolates Q412 and Q311 and aggressive isolates VA and CESS16K of O. ulmi were grown on solid medium, treated with QP1 antibodies, labelled with protein A – gold, and prepared for scanning electron microscopy. The gold-labelled QP1 polypeptide was detected on the leading edge of a small number of hyphae from nonaggressive isolates Q412 and Q311. Key words: immunogold labelling, Ophiostoma ulmi, soluble proteins.

scholarly journals Immunogold study on lectin binding in the porcine zona pellucida and granulosa cells

10.4081/846 ◽  
2009 ◽  
Vol 47 (4) ◽  
pp. 353 ◽  
Author(s):  
F Parillo ◽  
C Dall’Aglio ◽  
A Verini Supplizi ◽  
P Ceccarelli ◽  
AM Gargiulo
Keyword(s):  
Sialic Acid ◽  
Horseradish Peroxidase ◽  
Granulosa Cells ◽  
Zona Pellucida ◽  
Lectin Binding ◽  
Protein A ◽  
Ultrastructural Localization ◽  
Binding Pattern ◽  
Con A ◽  
Lectin Receptors

An ultrastructural localization of lectin receptors on the zona pellucida (ZP) of porcine antral oocytes and on the granulosa cells was performed using a panel of horseradish peroxidase- labelled lectins in conjunction with antiperoxidase antibody and protein A-gold. In some cases, lectin incubation was preceded by sialidase digestion. WGA-, Con-A-, UEA-I-, RCA-I-, PNA- and SBA-reactive sites were distributed differently in the porcine ZP. Sialidase digestion increased the positivity obtained with RCA-I and it was necessary to promote PNA and SBA reactivity. These results indicated that the ZP contained N-acetylglucosamine, a-mannose, a- fucose, b-Gal-(1-4)GlcNAc, b-Gal- (1-3)GalNAc, b-GalNAc and sialic acid residues. We also observed the presence of vesicles in both the ooplasm and granulosa cells, showing a similar lectin binding pattern to that of the ZP, thus suggesting that the oocyte and granulosa cells are the site of synthesis of ZP glucidic determinants.

scholarly journals Visualization of anti-sperm plasma membrane IgG and Fab as a method for localization of boar sperm membrane antigens.

1982 ◽  
Vol 30 (12) ◽  
pp. 1217-1227 ◽  
Author(s):  
L D Russell ◽  
R N Peterson ◽  
T A Russell
Keyword(s):  
Plasma Membrane ◽  
Protein A ◽  
Fluorescein Isothiocyanate ◽  
Ultrastructural Localization ◽  
Surface Antigens ◽  
Ultrastructural Level ◽  
Simple Method ◽  
Sperm Surface ◽  
Sperm Plasma Membrane ◽  
Sperm Antigens

A simple method for ultrastructural localization of sperm surface antigens by direct visualization of bound antibodies is presented. Anti-sperm plasma membrane (ASPM) immunoglobulin (Ig) G, visualized in tissues treated with an osmium:ferrocyanide mixture, projected 11-13 nm from the surface and ASPM Fab fragments projected 8-10 nm from the surface. The density of IgG labeling, as subjectively estimated, corresponded to indirect immune fluorescein isothiocyanate, indirect immunoferritin, and sperm-vesicle labeling patterns. Agglutination of sperm vesicles and sperm were demonstrated and the linking antibody visualized. A second antibody on protein A directed against ASPM IgG made the immunologic tag more apparent and indicated, in disrupted sperm preparations, labeling of both sides of the plasma membrane. The method provides for easy and sensitive localization of sperm surface antigens at the ultrastructural level and is presently being used to localize specific sperm antigens.

scholarly journals Immunoelectron microscopic demonstration of S-100b protein-like in centriole, cilia, and basal body.

1988 ◽  
Vol 36 (6) ◽  
pp. 693-696 ◽  
Author(s):  
T Uchida ◽  
T Endo
Keyword(s):  
Basal Body ◽  
Colloidal Gold ◽  
Protein A ◽  
Ultrastructural Localization ◽  
Basal Bodies ◽  
Cell Organelles ◽  
Gold Particles ◽  
Microscopic Demonstration ◽  
S 100 ◽  
And Function

We report here the ultrastructural localization of S-100b protein-like immunoreactivity in the centriole, cilia, and basal body. Duodenum and trachea of guinea pigs and rats were fixed and immunostained by the protein A-gold method. All centrioles, cilia, and basal bodies observed showed clear S-100b protein-like immunoreactivity. Specific colloidal gold particles were located over the microtubules in these cell organelles. However, other microtubules scattered throughout the cytoplasm were devoid of immunoreactivity. Although the functional significance of S-100b protein-like immunoreactivity in the centriole, cilia, and basal bodies remains to be elucidated, the present results introduce new perspectives into the investigation of localization and function of S-100 proteins.

scholarly journals Use of the protein A-gold technique for the morphological study of vascular permeability.

1980 ◽  
Vol 28 (11) ◽  
pp. 1251-1254 ◽  
Author(s):  
M Bendayan
Keyword(s):  
Endothelial Cells ◽  
Vascular Permeability ◽  
Ultrastructural Study ◽  
Morphological Study ◽  
Protein A ◽  
Gold Complex ◽  
Buffer Solution ◽  
General Applicability ◽  
Immunocytochemical Technique ◽  
Tissue Sections

The protein A-gold immunocytochemical technique has been applied for the ultrastructural study of vascular permeability. A capillary preparation was perfused for 30 min with a buffer solution containing albumin. After fixation and embedding, the albumin molecules were revealed on the tissue sections using anti-albumin antiserum and the protein A-gold complex. A specific labeling was obtained over the capillary lumen, the vesicular structures of the endothelial cells, and the intercapillary space. These results allowed us to conclude that the protein A-gold technique is suitable and of general applicability for the morphological study of vascular permeability.

scholarly journals Quantitative immunocytochemical localization of Na+,K+-ATPase in rat ciliary epithelial cells.

1989 ◽  
Vol 37 (9) ◽  
pp. 1353-1361 ◽  
Author(s):  
T Okami ◽  
A Yamamoto ◽  
K Omori ◽  
M Akayama ◽  
M Uyama ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Aqueous Humor ◽  
Ciliary Body ◽  
Protein A ◽  
Ultrastructural Localization ◽  
Immunoblot Analysis ◽  
Alpha Subunit ◽  
Ciliary Epithelium ◽  
Immunocytochemical Localization ◽  
Gold Particles

Ultrastructural localization of Na+,K+-ATPase in rat ciliary epithelium was investigated quantitatively by the protein A-gold technique, using an affinity-purified antibody against the alpha-subunit of Na+,K+-ATPase. Immunoblot analysis showed that the antibody bound specifically to the alpha-subunit of Na+,K+-ATPase in the ciliary body. Gold particles were found mainly on the basolateral surfaces of both the pigmented epithelial (PE) and nonpigmented epithelial (NPE) cells with an approximately twofold higher labeling density in the PE cells. A few gold particles were also found on the apical and ciliary channel surfaces of the PE cells, whereas no significant binding was found on the apical surfaces of the NPE cells. The basolateral surfaces of PE and NPE cells are markedly infolded and are much greater in area than the apical surfaces. This means that Na+,K+-ATPase is almost exclusively located on the basolateral surfaces of both the NPE and PE cells. We suggest that the Na+,K+-ATPase of both the NPE and PE cells play an important role in the formation of aqueous humor.

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