scholarly journals Codistribution of collagen types IV and AB2 in basement membranes and mesangium of the kidney. an immunoferritin study of ultrathin frozen sections.

1980 ◽  
Vol 85 (3) ◽  
pp. 597-616 ◽  
Author(s):  
F J Roll ◽  
J A Madri ◽  
J Albert ◽  
H Furthmayr
Keyword(s):  
Ultrastructural Level ◽  
Basement Membranes ◽  
Frozen Sections ◽  
Mesangial Matrix ◽  
Collagen Types ◽  
Type Iv ◽  
Collagen Antibodies ◽  
Ultrathin Frozen Sections ◽  
Bowman's Capsule ◽  
Large Vessels

Affinity-purified rabbit antibodies specific for collagen types I, III, AB2 and for a partially characterized type IV collagen derived from a murine tumor were used to study the distribution of collagens in the normal mouse kidney. Immunofluorescence staining of conventional frozen sections demonstrated that types I and III were present in bundles around large vessels and in fibers surrounding glomeruli and tubules, whereas types IV and AB2 were distributed in a linear fashion along basement membranes of tubules, glomeruli, and Bowman's capsule and in the mesangial stalk. The distribution of types IV nd AB2 was examined at the ultrastructural level by staining of 600- to 800-A thick frozen sections with a three-stage procedure employing specific collagen antibodies, biotinyl sheep antirabbit IgG, and avidin-ferritin conjugates. Labeling by this procedure demonstrated codistribution of types AB2 and the putative type IV in all three basement membranes. In addition, mesangial matrix was shown to contain both of these collagen types. These results support recent biochemical evidence of collagen heterogeneity in basement membranes, and also support the concept of a structural relationship between mesangial matrix and glomerular basement membranes.

scholarly journals Ultrastructural localization of fibronectin and laminin in the basement membranes of the murine kidney.

1980 ◽  
Vol 86 (2) ◽  
pp. 682-687 ◽  
Author(s):  
J A Madri ◽  
F J Roll ◽  
H Furthmayr ◽  
J M Foidart
Keyword(s):  
Mesangial Cell ◽  
Ultrastructural Localization ◽  
Ultrastructural Level ◽  
Basement Membranes ◽  
Frozen Sections ◽  
Mesangial Matrix ◽  
Lamina Densa ◽  
Rabbit Igg ◽  
Cell Processes ◽  
Bowman's Capsule

Affinity-purified rabbit antibodies specific for two large noncollagenous gycoproteins--laminin and fibronectin--were used to study the distribution of these proteins in normal murine kidneys. Immunofluorescence staining of conventional frozen sections demonstrates fibronectin within mesangial areas of the glomerulus. Laminin is also found in mesangial areas. However, it also appears to be distributed in typical basement membranelike patterns on glomerular and tubular basement membranes and Bowman's capsule. At the ultrastructural level, by labeling 600-800-A thick frozen sections with a three-stage procedure consisting of specific antibodies, biotinyl sheep anti-rabbit IgG, and avidin-ferritin conjugates, fibronectin is present ony in the mesangial matrix and is specifically localized to areas immediately surrounding mesangial cell processes. Laminin, on the other hand, is found uniformly distributed throughout tubular basement membranes, the mesangial matrix, and Bowman's capsule. In glomerular basement membranes, laminin labeling is restricted to the lamina rara interna and adjacent regions of the lamina densa.

Immunohistochemical Localization of Collagen Types I, III, and IV, Laminin, Fibronectin, and Keratin in the Endolymphatic Sac

2000 ◽  
Vol 109 (2) ◽  
pp. 180-186 ◽  
Author(s):  
Teruhiko Harada ◽  
Youngki Kim ◽  
Steven K. Juhn ◽  
Yasuo Sakakura
Keyword(s):  
Collagen Type ◽  
Collagen Type I ◽  
Immunohistochemical Localization ◽  
Basement Membranes ◽  
Endolymphatic Sac ◽  
Type I ◽  
Collagen Types ◽  
Type Iv ◽  
Baseline Information ◽  
Subepithelial Layer

We have employed immunohistochemistry to obtain baseline information on the molecular constituents of the extracellular matrix (ECM) of the endolymphatic duct (ED) and endolymphatic sac (ES) of the chinchilla. The results demonstrated that collagen types I and III were distributed in the subepithelial layer in the ED and ES, type IV collagen and laminin in the basement membranes, and fibronectin in the subepithelial layer and partly in the conglomerated cells in the ES. Collagen type III was diffusely distributed in the whole subepithelial layer of the ES, whereas collagen type I was concentrated densely in the deep layer of the interstitium, although gradually, the cuboidal epithelium in the ES was transformed into a flatter type in the ED. The epithelial cells of the ED and ES were clearly positive for keratin. This study deals, in particular, with the normal distribution of ECM components of the ED and ES of the chinchilla.

scholarly journals Secretion and Assembly of Type IV and VI Collagens Depend on Glycosylation of Hydroxylysines

2007 ◽  
Vol 282 (46) ◽  
pp. 33381-33388 ◽  
Author(s):  
Laura Sipilä ◽  
Heli Ruotsalainen ◽  
Raija Sormunen ◽  
Naomi L. Baker ◽  
Shireen R. Lamandé ◽  
...  
Keyword(s):  
Congenital Muscular Dystrophy ◽  
Basement Membranes ◽  
Collagen Vi ◽  
Collagen Types ◽  
Type Vi Collagen ◽  
Knock Out ◽  
Type Iv ◽  
New Information ◽  
Ullrich Congenital Muscular Dystrophy

Most lysines in type IV and VI collagens are hydroxylated and glycosylated, but the functions of these unique galactosylhydroxylysyl and glucosylgalactosylhydroxylysyl residues are poorly understood. The formation of glycosylated hydroxylysines is catalyzed by multifunctional lysyl hydroxylase 3 (LH3) in vivo, and we have used LH3-manipulated mice and cells as models to study the function of these carbohydrates. These hydroxylysine-linked carbohydrates were shown recently to be indispensable for the formation of basement membranes (Ruotsalainen, H., Sipilä, L., Vapola, M., Sormunen, R., Salo, A. M., Uitto, L., Mercer, D. K., Robins, S. P., Risteli, M., Aszodi, A., Fässler, R., and Myllylä, R. (2006) J. Cell Sci. 119, 625–635). Analysis of LH3 knock-out embryos and cells in this work indicated that loss of glycosylated hydroxylysines prevents the intracellular tetramerization of type VI collagen and leads to impaired secretion of type IV and VI collagens. Mice lacking the LH activity of LH3 produced slightly underglycosylated type IV and VI collagens with abnormal distribution. The altered distribution and aggregation of type VI collagen led to similar ultrastructural alterations in muscle to those detected in collagen VI knockout and some Ullrich congenital muscular dystrophy patients. Our results provide new information about the function of hydroxylysine-linked carbohydrates of collagens, indicating that they play an important role in the secretion, assembly, and distribution of highly glycosylated collagen types.

scholarly journals Characterization of the Type 3 Fimbrial Adhesins ofKlebsiella Strains

1998 ◽  
Vol 66 (6) ◽  
pp. 2887-2894 ◽  
Author(s):  
Tricia A. Schurtz Sebghati ◽  
Timo K. Korhonen ◽  
Douglas B. Hornick ◽  
Steven Clegg
Keyword(s):  
Basement Membranes ◽  
Collagen Types ◽  
Type V Collagen ◽  
Type Iv ◽  
Fimbrial Adhesins ◽  
Type V ◽  
Type 3 ◽  
Fimbrial Adhesin

ABSTRACT The Klebsiella pneumoniae fimbrial adhesin, MrkD, mediates adherence to the basolateral surfaces of renal and pulmonary epithelia and to the basement membranes of tissues. Although all isolates possessing the MrkD adhesin mediate the agglutination, in vitro, of erythrocytes treated with tannic acid, the mrkDgene is not conserved within species. The ability of a plasmid-bornemrkD gene product to mediate binding to type V collagen is associated frequently with strains of K. oxytoca and rarely with strains of K. pneumoniae. In K. pneumoniae, the MrkD adhesin is located within a chromosomally borne gene cluster and mediates binding to collagen types IV and V. The plasmid-borne determinant, mrkD 1P, and the chromosomally borne gene, mrkD 1C, are not genetically related. Some strains of enterobacteria possess amrkD 1C allele that is associated with hemagglutinating activity but does not bind to either type IV or type V collagen.

scholarly journals Fibronectin localization in the rat glomerulus.

1980 ◽  
Vol 87 (3) ◽  
pp. 691-696 ◽  
Author(s):  
P J Courtoy ◽  
Y S Kanwar ◽  
R O Hynes ◽  
M G Farquhar
Keyword(s):  
Epithelial Cells ◽  
Mesangial Cell ◽  
Cultured Cells ◽  
Mesangial Cells ◽  
Basement Membranes ◽  
Frozen Sections ◽  
Mesangial Matrix ◽  
Capillary Loop ◽  
Important Constituent

Fibronectin (FN) has been localized in the rat glomerulus using indirect immunolabeling. It was demonstrated in frozen sections by immunofluorescence, in sections of fixed kidneys by both peroxidase and ferritin-labeled antibodies, and in isolated glomerular basement membranes (GBM) with ferritin-labeled antibodies. Complementary and convergent results were obtained with these approaches. FN was most abundant in the mesangial matrix where it was especially concentrated at the interface between the endothelial and mesangial cells. In the peripheral capillary loop, FN was also detected in the laminae rarae (interna and externa) of the GBM--i.e., between the endothelial and epithelial cells, respectively, and the GBM. These findings indicate that FN is an important constituent of the glomerulus, and they are compatible with the assumption that, in the glomerulus, as in cultured cells, FN is involved in cell-to-cell (mesangial-mesangial, mesangial-endothelial) and cell-to-substrate (mesangial cell-mesangial matrix, epithelium-GBM, endothelium-GBM) attachment.

scholarly journals Immunoelectronmicroscopy of neural antigens on ultrathin frozen sections.

1986 ◽  
Vol 34 (4) ◽  
pp. 491-500 ◽  
Author(s):  
M R Celio ◽  
G A Keller ◽  
F E Bloom
Keyword(s):  
High Resolution ◽  
Organic Solvents ◽  
Ultrastructural Level ◽  
Frozen Sections ◽  
Neuronal Function ◽  
Enzyme Marker ◽  
Ultrathin Frozen Sections

We have utilized immunocryoultramicrotomy to detect synapsin, somatostatin, parvalbumin, and tubulin at the ultrastructural level. Immunocryoultramicrotomy combines informative identification of morphology with accurate immunolabeling. Moreover, since no detergents or organic solvents are used to enable antibody penetration, and since no enzyme marker diffusion occurs, localization of the antigens should be more accurate. Accordingly, it was possible to localize precisely all four antigens within a well-preserved structure. Application of this method has important advantages for high-resolution localization of molecules relevant to neuronal function.

scholarly journals Human Basement Membrane Collagens do not Elicit Platelet Aggregation

1977 ◽  
Author(s):  
R. L. Trelstad ◽  
A. C. Carvalho
Keyword(s):  
Platelet Aggregation ◽  
Basement Membrane ◽  
Collagen Type ◽  
Serotonin Release ◽  
Basement Membranes ◽  
Collagen Type Iv ◽  
Collagen Types ◽  
Type Iv ◽  
Skin Collagen ◽  
Human Collagen

The immediate subendothelial connective tissue matrix consists of the basement membrane, a collagenous felt-like cell surface coat. The collagen from basement membranes has been isolated from human lung, skin, and kidney using a new fractionation method which separates native forms of collagen Types I, II, III, and IV. The Type IV collagens from the basement membranes have been characterized in respect to amino acid and carbohydrate composition, molecular size, charge and native structure. Antibodies prepared against the Type IV collagen reacted with both epithelial and vascular basement membranes as judged by immunofluorescence. Platelet-rich plasma (250,000/μl) from 5 normal subjects were tested for aggregation and 14C-serotonin release with human collagen Types I, II, III, and IV. Complete aggregation (100%) and 14C-serotonin release (80–100%) was obtained when Types I, II, and III were used. Human kidney, lung, and skin collagen Type IV (10–100μg/ml) did not aggregate platelets nor cause release of their contents. Pre-incubation of platelets and human collagen Type IV for periods of 30 minutes did not result in inhibition of platelet aggregation by Types I, II, or III.These data indicate that the collagenous component of the basement membrane, the first extra-vascular collagen to which a platelet is exposed, does not elicit aggregation as do the fibrillar collagens in the perivascular matrix.

scholarly journals Connective tissue of rat lung. II: Ultrastructural localization of collagen types III, IV, and VI.

1988 ◽  
Vol 36 (9) ◽  
pp. 1167-1173 ◽  
Author(s):  
P S Amenta ◽  
J Gil ◽  
A Martinez-Hernandez
Keyword(s):  
Type I Collagen ◽  
Ultrastructural Localization ◽  
Basement Membranes ◽  
Type Iii Collagen ◽  
Similar Distribution ◽  
Type I ◽  
Rat Lung ◽  
Collagen Types ◽  
Type Iii ◽  
Type Iv

We localized collagen types III, IV, and VI in normal rat lung by light and electron immunohistochemistry. Type IV collagen was present in every basement membrane examined and was absent from all other structures. Although types III and VI had a similar distribution, being present in the interstitium of major airways, blood vessels, and alveolar septa, as in other organs, they had different morphologies. Type III collagen formed beaded fibers, 15-20 nm in diameter, whereas type VI collagen formed fine filaments, 5-10 nm in diameter. Both collagen types were found exclusively in the interstitium, often associated with thick (30-35 nm) cross-banded type I collagen fibers. Occasionally, type III fibers and type VI filaments could be found bridging from the interstitium to the adventitial aspect of some basement membranes. Furthermore, the association of collagen type VI with types I and III and basement membranes suggests that type VI may contribute to integration of the various components of the pulmonary extracellular matrix into a functional unit.

scholarly journals Comparative distribution of laminin, type IV collagen, and fibronectin in the rat glomerulus.

1982 ◽  
Vol 30 (9) ◽  
pp. 874-886 ◽  
Author(s):  
P J Courtoy ◽  
R Timpl ◽  
M G Farquhar
Keyword(s):  
Type Iv Collagen ◽  
Rat Kidney ◽  
Basement Membranes ◽  
Kidney Cortex ◽  
Mesangial Matrix ◽  
Lamina Densa ◽  
Rat Kidney Cortex ◽  
Type Iv ◽  
Attachment Proteins ◽  
Cryostat Sections

The distribution of laminin, type IV collagen, and fibronectin was investigated in the rat kidney cortex by immunolabeling. It was demonstrated by immunofluorescence on both unfixed cryostat sections and fixed ultracryomicrotome sections, by immunoperoxidase on fixed cryostat sections, and by immunoferritin on isolated glomerular basement membranes (GBM). This multifaceted approach provided complementary and convergent results. Distinct patterns were found for each antigen in the glomerulus and remaining kidney cortex. Laminin was localized predominantly in the GBM, where it was concentrated in the laminae rarae. Staining also occurred to a lesser extent in the mesangial matrix. Type IV collagen was evenly distributed in the lamina densa of the GBM and in the mesangial matrix. Fibronectin was most abundant in the mesangial matrix, but it could also be detected in the peripheral GBM, where it was localized in the laminae rarae. Labeling for fibronectin was particularly prominent at the endothelial-mesangial interface. The findings indicate that the three layers of the GBM differ in their composition: The lamina densa contains type IV collagen and the laminae rarae contain the two attachment proteins, fibronectin and laminin. The mesangial matrix stains for all three antigens, but it is also heterogeneous and can be subdivided into several domains--i.e., the endothelial-mesangial matrix, which is particularly rich in fibronectin, the intermesangial matrix, which contains mainly type IV collagen and fibronectin, and the GBM (where it continues over the mesangial regions), which stains most heavily for laminin.

Immunoferritin localization of intracellular antigens in positively stained frozen sections

1978 ◽  
Vol 36 (2) ◽  
pp. 168-169
Author(s):  
K. T. Tokuyasu
Keyword(s):  
Surface Tension ◽  
Water Surface ◽  
Thin Layers ◽  
The Other ◽  
Positive Staining ◽  
Frozen Sections ◽  
The Past ◽  
Ultrathin Frozen Sections ◽  
Definition Of

During the past investigations of immunoferritin localization of intracellular antigens in ultrathin frozen sections, we found that the degree of negative staining required to delineate u1trastructural details was often too dense for the recognition of ferritin particles. The quality of positive staining of ultrathin frozen sections, on the other hand, has generally been far inferior to that attainable in conventional plastic embedded sections, particularly in the definition of membranes. As we discussed before, a main cause of this difficulty seemed to be the vulnerability of frozen sections to the damaging effects of air-water surface tension at the time of drying of the sections.Indeed, we found that the quality of positive staining is greatly improved when positively stained frozen sections are protected against the effects of surface tension by embedding them in thin layers of mechanically stable materials at the time of drying (unpublished).

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