The use of 1nm silver enhanced gold probes for the detection of skin antigens

1990 ◽  
Vol 48 (3) ◽  
pp. 902-903
Author(s):  
J.P Cassella ◽  
H. Shimizu ◽  
A. Ishida-Yamamoto ◽  
R.A.J. Eady
Keyword(s):  
Type Iv Collagen ◽  
Freeze Substitution ◽  
Collagen Type Iv ◽  
Electron Microscopic ◽  
Normal Human Skin ◽  
Type Iv ◽  
Type Vii Collagen ◽  
Keratin Filaments ◽  
Normal Human ◽  
Phosphate Buffered Saline

1nm colloidal gold with silver enhancement has been used in conjunction with a low-temperature post-embedding (post-E) technique for the demonstration of skin antigens at both the light microscopic (LM) and electron microscopic (EM) levels.Keratin filaments and basement membrane zone (BMZ) associated antigens in normal human skin (NHS) were immunolabelled using antibodies against keratin 14, 10, and 1, the carboxy-terminus and collagenous portion of type VII collagen, type IV collagen and bullous pemphigoid antigen (BP-Ag).Fresh samples of NHS were cryoprotected in 15% glycerol, cryofixed in propane at -190°C, subjected to freeze substitution in methanol at -80°C and embedded in Lowicryl K11M at -60°C. Polymerisation of the resin was initiated under UVR at - 60°C for 48 hours and continued at room temperature for a further 48 hours. Semith in sections were air dried onto slides coated with 3-aminopropyltriethoxysilane. The following immunolabelling protocol was adopted: Primary antibody was applied for 2 hours at 37°C or overnight at 4°C. Following washing in Dulbecco’s phosphate buffered saline (PBSA) a biotinylated secondary antibody was applied for 2 hours at 37°C. The sections were further washed in PBSA and 1nm gold avidin was applied. Sections were finally washed in PBSA and silver enhanced.

Immunohistochemical localization of type IV collagen in normal human skin.

1986 ◽  
Vol 48 (5) ◽  
pp. 920-923
Author(s):  
Hiromi KOBAYASHI ◽  
Masamitsu ISHII ◽  
Miyako CHANOKI ◽  
Kazuyoshi FUKAI ◽  
Toshio HAMADA ◽  
...  
Keyword(s):  
Human Skin ◽  
Type Iv Collagen ◽  
Immunohistochemical Localization ◽  
Normal Human Skin ◽  
Type Iv ◽  
Normal Human

The Subbasement Membrane Distribution of Type IV Collagen in Normal Human Skin

1989 ◽  
Vol 16 (6) ◽  
pp. 458-463 ◽  
Author(s):  
Hiromi Kobayashi ◽  
Masamitsu Ishii ◽  
Miyako Chanoki ◽  
Kazuyoshi Fukai ◽  
Toshio Hamada ◽  
...  
Keyword(s):  
Human Skin ◽  
Type Iv Collagen ◽  
Normal Human Skin ◽  
Type Iv ◽  
Normal Human

scholarly journals Immunoelectron Microscopic Characterization of Human Dermal Lymphatic Microvascular Endothelial Cells: Differential Expression of CD31, CD34, and Type IV Collagen with Lymphatic Endothelial Cells vs Blood Capillary Endothelial Cells in Normal Human Skin, Lymphangioma, and Hemangioma In Situ

1998 ◽  
Vol 46 (2) ◽  
pp. 165-176 ◽  
Author(s):  
Birthe Sauter ◽  
Dagmar Foedinger ◽  
Barbara Sterniczky ◽  
Klaus Wolff ◽  
Klemens Rappersberger
Keyword(s):  
Endothelial Cells ◽  
Human Skin ◽  
Type Iv Collagen ◽  
Microvascular Endothelial Cells ◽  
Lymphatic Endothelial Cells ◽  
Normal Human Skin ◽  
Type Iv ◽  
Normal Human ◽  
Microvascular Endothelial

We performed a comparative investigation of the immunomorphological characteristics of lymphatic and blood microvascular endothelial cells in normal human skin, cutaneous lymphangiomas, and hemangiomas, employing a pre-embedding immunogold electron microscopic technique. We stained for cell membrane proteins that are commonly used for light microscopic characterization of blood endothelial cells. With blood microvascular endothelial cells, we observed uniform labeling of the luminal cell membranes with monoclonal antibodies (MAbs) JC70 (CD31), EN-4 (CD31), BMA120, PAL-E, and QBEND-10 (CD34), and strong staining of the vascular basal lamina for Type IV collagen under normal and pathological conditions. In contrast, lymphatic microvascular endothelial cells in normal human skin and in lymphangiomas displayed, in addition to a luminal labeling, pronounced expression of CD31 and CD34 along the abluminal cell membranes. Moreover, CD31 was preferentially detected within intercellular junctions. The expression of CD34 was mostly confined to abluminal endothelial microprocesses and was upregulated in lymphangiomas and hemangiomas. Type IV collagen partially formed the luminal lining of initial lymphatics and occasionally formed bridges over interendothelial gaps. Our findings suggest a function of transmigration protein CD31 in recruitment of dendritic cells into the lymphatic vasculature. CD34 labeling may indicate early endothelial cell sprouting. The distribution of Type IV collagen also supports its role as a signal for migration and tube formation for lymphatic endothelial cells.

scholarly journals Monoclonal antibodies against chicken type IV and V collagens: electron microscopic mapping of the epitopes after rotary shadowing.

1984 ◽  
Vol 98 (5) ◽  
pp. 1637-1644 ◽  
Author(s):  
R Mayne ◽  
H Wiedemann ◽  
M H Irwin ◽  
R D Sanderson ◽  
J M Fitch ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Cleavage Site ◽  
Type Iv Collagen ◽  
Collagen Molecule ◽  
Electron Microscopic ◽  
Type V Collagen ◽  
Type Iv ◽  
Type V ◽  
Rotary Shadowing

The location of the epitopes for monoclonal antibodies against chicken type IV and type V collagens were directly determined in the electron microscope after rotary shadowing of antibody/collagen mixtures. Three monoclonal antibodies against type IV collagen were examined, each one of which was previously demonstrated to be specific for only one of the three pepsin-resistant fragments of the molecule. The three native fragments were designated (F1)2F2, F3, and 7S, and the antibodies that specifically recognize each fragment were called, respectively, IA8 , IIB12 , and ID2 . By electron microscopy, monoclonal antibody IA8 recognized an epitope located in the center of fragment (F1)2F2 and in tetramers of type IV collagen at a distance of 288 nm from the 7S domain, the region of overlap of four type IV molecules. Monoclonal antibody IIB12 , in contrast, recognized an epitope located only 73 nm from the 7S domain. This result therefore provides direct visual evidence that the F3 fragment is located closest to the 7S domain and the order of the fragments must be 7S-F3-(F1)2F2. The epitope for antibody ID2 was located in the overlap region of the 7S domain, and often several antibody molecules were observed to binding to a single 7S domain. The high frequency with which antibody molecules were observed to bind to fragments of type IV collagen suggests that there is a single population of type IV molecules of chain organization [alpha 1(IV)]2 alpha 2(IV), and that four identical molecules must form a tetramer that is joined in an antiparallel manner at the 7S domain. The monoclonal antibodies against type V collagen, called AB12 and DH2 , were both found to recognize epitopes close to one another, the epitopes being located 45-48 nm from one end of the type V collagen molecule. The significance of this result still remains uncertain, but suggests that this site is probably highly immunoreactive. It may also be related to the specific cleavage site of type V collagen by selected metalloproteinases and by alpha-thrombin. This cleavage site is also known to be located close to one end of the type V molecule.

scholarly journals Expression of collagen type IV in human kidney during prenatal development

2020 ◽  
pp. 111-111
Author(s):  
Vladimir Petrovic ◽  
Ivan Nikolic ◽  
Marko Jovic ◽  
Vladimir Zivkovic ◽  
Miodrag Jocic ◽  
...  
Keyword(s):  
Type Iv Collagen ◽  
Collagen Type ◽  
Kidney Tissue ◽  
Volume Density ◽  
Collagen Iv ◽  
Basement Membranes ◽  
Collagen Type Iv ◽  
Type Iv ◽  
Metanephric Kidney ◽  
Collecting System

Background / Aim. Type IV collagen belongs to the group of non-fibrillar collagens and is an important component of the basement membranes where it accounts for approximately 50% of its structural elements. The aim of the paper was to describe the expression and distribution of collagen type IV in embryonic and fetal metanephric kidney, and to determine the volume density of collagen type IV in kidney tissue in each trimester of development. Methods. The material consisted of 19 human embryos/fetuses, in the gestational age from 8th to 37th week. Kidney tissue specimens were routinely processed to paraffin molds and stained with hematoxylin and eosin and immunohistochemically using polyclonal anti-collagen IV antibody. Stained slides were examined using light microscope and images of the selected areas, under different lens magnification were captured with digital camera. Volume density of collagen type IV was determined by using ImageJ 1.48v and a plugin of the software which inserted a grid system with 336 points. For the data comparison One-Way Analysis of Variance was used. Results. Strong collagen IV immunopositivity was seen in all specimens, with a distribution in the basement membranes of urinary bud, parietal leaf of Bowman?s capsule, glomerular basement membrane, basement membrane of interstitial blood vessels, and basement membranes of nephron tubules and collecting ducts. No statistically significant difference in the volume density of type IV collagen was found between the different trimesters of development. Conclusion. The synthesis and secretion of collagen type IV simultaneously follows the development of nephron structures, collecting system and blood vessels. The volume density of collagen type IV remains constant throughout all the trimesters of metanephric kidney development, indicating that it plays a crucial role in normal development of nephron and collecting system structures, as well as in maintaining the normal kidney function.

Malotilate Prevents Accumulation of Type III pN-Collagen, Type IV Collagen, and Laminin in Carbon Tetrachloride-induced Pulmonary Fibrosis in Rats

1989 ◽  
Vol 139 (5) ◽  
pp. 1105-1111 ◽  
Author(s):  
Paavo Pääkkö ◽  
Raija Sormunen ◽  
Leila Risteli ◽  
Juha Risteli ◽  
Leena Ala-Kokko ◽  
...  
Keyword(s):  
Carbon Tetrachloride ◽  
Pulmonary Fibrosis ◽  
Type Iv Collagen ◽  
Collagen Type ◽  
Collagen Type Iv ◽  
Type Iii ◽  
Type Iv

Extracellular matrix components in the heart of the white bass, Morone chrysops (Rafinesque)

1994 ◽  
Vol 72 (3) ◽  
pp. 449-454
Author(s):  
Dominic S. Raso ◽  
Louis Terracio ◽  
Thomas K. Borg
Keyword(s):  
Type Iv Collagen ◽  
Collagen Type ◽  
Periodic Acid ◽  
Longitudinal Axis ◽  
Collagen Type Iv ◽  
Histological Techniques ◽  
White Bass ◽  
Morone Chrysops ◽  
Type Iv ◽  
Extracellular Matrix Components

The distribution of laminin, collagen type IV, collagen bundles, proteoglycans, elastin, and periodic acid–Schiff's moieties (glycoproteins) within the heart of the adult white bass, Morone chrysops (Rafinesque), was investigated by means of immunohistochemical and histological techniques. Laminin and collagen type IV were heavily expressed within the epimysium and the basal lamina of the lining epicardial epithelium and valvular endothelium, moderately expressed within the myocardium, and slightly expressed within the subendocardium. This co-localized distribution of laminin and collagen type IV corresponds to the biochemically unidentified basal and external lamina observed in the hearts of other fish by previous ultrastructural investigations and is similar to the distribution observed in the hearts of birds and mammals. Also demonstrated was an interesting division of connective tissue components along the longitudinal axis of the atrioventricular valve, which is most likely intimately involved with the effective functioning and durability of the valve.

Malignant Peripheral Nerve Sheath Tumor With a t(X;18)

2000 ◽  
Vol 124 (6) ◽  
pp. 864-867 ◽  
Author(s):  
Russell Vang ◽  
David A. Biddle ◽  
Wilbur R. Harrison ◽  
Kent Heck ◽  
Linda D. Cooley
Keyword(s):  
Peripheral Nerve ◽  
Synovial Sarcoma ◽  
Chromosomal Translocation ◽  
Collagen Type Iv ◽  
Variant Form ◽  
Nerve Sheath Tumor ◽  
Electron Microscopic ◽  
Peripheral Nerve Sheath Tumor ◽  
Type Iv ◽  
Nerve Sheath

Abstract We describe an ankle tumor arising in a 16-year-old girl. The tumor demonstrated histology typical of a malignant peripheral nerve sheath tumor (MPNST), but exhibited a variant form of the (X;18) translocation associated with synovial sarcoma. Immunohistochemical stains were positive for vimentin, CD57, collagen type IV, and Bcl-2. Routine and molecular cytogenetic studies showed an unbalanced 3-way chromosomal translocation that involved chromosomes X, 18, and 1. Electron microscopic findings were noncontributory. This unusual tumor raises the following questions and possibilities: (1) As the t(X;18) suggests, could this tumor be a monophasic synovial sarcoma with the histologic features of an MPNST? (2) Or, as the histology suggests, is this tumor an MPNST that has a t(X;18)? (3) Finally, could MPNST histology, a t(X;18), and no defining immunohistochemical or electron microscopic features represent an as yet unrecognized part of a spectrum that spans from synovial sarcoma to MPNST or other spindle cell tumors?

scholarly journals Antibodies to basement membrane collagen and to laminin are present in sera from patients with poststreptococcal glomerulonephritis.

1986 ◽  
Vol 163 (3) ◽  
pp. 588-602 ◽  
Author(s):  
N A Kefalides ◽  
M T Pegg ◽  
N Ohno ◽  
T Poon-King ◽  
J Zabriskie ◽  
...  
Keyword(s):  
Basement Membrane ◽  
Type Iv Collagen ◽  
Gel Filtration ◽  
Collagen Type Iv ◽  
Poststreptococcal Glomerulonephritis ◽  
Molecule Reaction ◽  
Goodpasture Syndrome ◽  
Type Iv ◽  
Anterior Lens Capsule ◽  
Collagenous Domain

Sera from patients with poststreptococcal glomerulonephritis (PSGN) known to have antibodies to proteoglycans were studied for the presence of antibodies against other basement membrane (BM) components. BM collagen (type IV) was isolated in the native state by extracting bovine anterior lens capsule (ALC) with 0.5 M acetic acid. The 7-S (collagenous) domain and the NC-1 (noncollagenous) domain of type IV collagen were obtained after bacterial collagenase digestion of ALC followed by gel filtration. Laminin was isolated from the mouse EHS tumor and fibronectin from human plasma. Immunologic studies, using an ELISA and electroimmunoblot, revealed the presence of antibodies that reacted with intact, native type IV collagen and the 7-S collagenous domain of this molecule. Reaction with the NC-1 (noncollagenous) domain was minimal, and not higher than that obtained with control sera. Laminin reaction strongly with the patients' sera, but fibronectin did not. Unlike sera from patients with Goodpasture syndrome, which contain antibodies primarily against the NC-1 (noncollagenous) domain of type IV collagen, sera from patients with acute PSGN contain antibodies against all the major macromolecular components of BM. This difference in immunologic reactivity may account for the observed differences in the pathologic picture at the glomerular level.

Sign in / Sign up

Export Citation Format

Share Document