scholarly journals THE USE OF BEEF LIVER CATALASE AS A PROTEIN TRACER FOR ELECTRON MICROSCOPY

1969 ◽  
Vol 42 (2) ◽  
pp. 480-489 ◽  
Author(s):  
M. A. Venkatachalam ◽  
H. Dariush Fahimi
Keyword(s):  
Ultrastructural Localization ◽  
Basement Membranes ◽  
Liver Uptake ◽  
Beef Liver ◽  
Barium Peroxide ◽  
Pinocytotic Vesicles ◽  
Intercellular Clefts ◽  
Urinary Space ◽  
Sensitive Method ◽  
Glomerular Capillaries

Beef liver catalase was injected intravenously into mice, and its distribution in the kidney, myocardium, and liver was studied with the electron microscope. A specific and relatively sensitive method was developed for its ultrastructural localization, based on the peroxidatic activity of catalase and employing a modified Graham and Karnovsky incubation medium. The main features of the medium were a higher concentration of diaminobenzidine, barium peroxide as the source of peroxide, and pH of 8.5. Ultrastructurally, the enzyme was seen to permeate the endothelial fenestrae and basement membranes of tubular and glomerular capillaries of the kidney. The urinary space and tubular lumina contained no reaction product. In the myocardial capillaries, the tracer filled the pinocytotic vesicles but did not diffuse across the intercellular clefts of the endothelium. In liver, uptake of catalase was seen both in hepatocytes and in Kupffer cells.

scholarly journals THE ULTRASTRUCTURAL BASIS OF ALVEOLAR-CAPILLARY MEMBRANE PERMEABILITY TO PEROXIDASE USED AS A TRACER

1968 ◽  
Vol 37 (3) ◽  
pp. 781-793 ◽  
Author(s):  
Eveline E. Schneeberger-Keeley ◽  
Morris J. Karnovsky
Keyword(s):  
Epithelial Cells ◽  
Protein Transport ◽  
Uranyl Acetate ◽  
Basement Membranes ◽  
Small Molecular Weight ◽  
Zonulae Occludentes ◽  
Capillary Membrane ◽  
Pinocytotic Vesicles ◽  
Intercellular Clefts ◽  
Alveolar Capillary

The permeability of the alveolar-capillary membrane to a small molecular weight protein, horseradish peroxidase (HRP), was investigated by means of ultrastructural cytochemistry. Mice were injected intravenously with HRP and sacrificed at varying intervals. Experiments with intranasally instilled HRP were also carried out. The tissue was fixed in formal-dehyde-glutaraldehyde fixative. Frozen sections were cut, incubated in Graham and Karnovsky's medium for demonstrating HRP activity, postfixed in OsO4, and processed for electron microscopy. 90 sec after injection, HRP had passed through endothelial junctions into underlying basement membranes, but was stopped from entering the alveolar space by zonulae occludentes between epithelial cells. HRP was demonstrated in pinocytotic vesicles of both endothelial and epithelial cells, but the role of these vesicles in net protein transport appeared to be minimal. Intranasally instilled HRP was similarly prevented from permeating the underlying basement membrane by epithelial zonulae occludentes. Pulmonary endothelial intercellular clefts stained with uranyl acetate appeared to contain maculae occludentes rather than zonulae occludentes. HRP did not alter the ultrastructure of these junctions.

scholarly journals GLOMERULAR PERMEABILITY

1969 ◽  
Vol 130 (2) ◽  
pp. 381-399 ◽  
Author(s):  
M. A. Venkatachalam ◽  
Morris J. Karnovsky ◽  
Ramzi S. Cotran
Keyword(s):  
Epithelial Cells ◽  
Basement Membrane ◽  
Intercellular Junctions ◽  
Ultrastructural Localization ◽  
Basement Membranes ◽  
The Other ◽  
Serial Sections ◽  
Renal Glomerulus ◽  
Glomerular Permeability ◽  
Urinary Space

Wistar/Furth rats were made nephrotic by daily administration of amino-nucleoside of puromycin, and the ultrastructural localization of horseradish peroxidase (mol wt 40,000) in the renal glomerulus was studied from 1 min to 20 hr after intravenous injection of the tracer. In control rats, peroxidase permeated the endothelial fenestrae, the basement membrane, and the epithelial slits, and was present in tubular lumina. Nephrotic glomeruli showed relatively normal basement membranes, extensive fusion of foot processes with formation of "close" intercellular junctions, and large vacuoles and pockets in epithelial cells. On serial sections some of the epithelial vacuoles communicated on one side with the extracellular space overlying basement membrane, and on the other side with the urinary space. In nephrotic animals, peroxidase permeated the basement membrane and the close junctions, and was present in many of the vacuoles and pockets as early as 1 min after injection. Only small numbers of peroxidase-positive vacuoles remained in. epithelial cells 1 hr or more after injection of the tracer. It is suggested that the epithelial pockets and vacuoles form pathways across which leaking proteins can be transferred across the epithelium into the urinary space. Epithelial vacuoles may also be absorption droplets designed to "conserve" leaking proteins, but this function was not prominent in our experiments with peroxidase.

scholarly journals Ultrastructural localization of type V collagen in rat kidney.

1982 ◽  
Vol 92 (2) ◽  
pp. 343-349 ◽  
Author(s):  
A Martinez-Hernandez ◽  
S Gay ◽  
E J Miller
Keyword(s):  
Type I Collagen ◽  
Rat Kidney ◽  
Ultrastructural Localization ◽  
Particulate Material ◽  
Basement Membranes ◽  
Type I ◽  
Type V Collagen ◽  
Collagen Molecules ◽  
Immunohistochemical Studies ◽  
Type V

Antibodies specific for the alpha 1 (V) chain and native collagen molecules containing the alpha 1 (V) chain have been used in electron immunohistochemical studies of rat kidney to determine the ultrastructural distribution of this class of collagen molecules. In addition, antibodies against type I collagen and whole basement membrane were used as markers for interstitial collagen and authentic basement membranes. Our results indicate that type V collagen is present in the renal interstitium in different forms: in close apposition to interstitial collagen fibers; in the stromal aspect of vascular basement membranes; and as particulate material not bound to other structures. On the basis of these findings, we postulate a binding or connecting function for this collagen type.

Glomerulosclerosis in Canine Heartworm Infection

1974 ◽  
Vol 11 (6) ◽  
pp. 506-514 ◽  
Author(s):  
C. F. Simpson ◽  
B. M. Gebhardt ◽  
R. E. Bradley ◽  
R. F. Jackson
Keyword(s):  
Electron Microscopy ◽  
Endothelial Cells ◽  
Immune Complexes ◽  
Tissue Damage ◽  
Basement Membranes ◽  
Canine Heartworm ◽  
Heartworm Disease ◽  
Heartworm Infection ◽  
Glomerular Lesions ◽  
Glomerular Capillaries

The kidneys of seven dogs with natural infections of heartworm disease, four dogs with experimental infections, and six control (uninfected) dogs raised in isolation were examined by light, fluorescent, and electron microscopy. Swelling and fragmentation of the basement membranes accompanied by denudations of endothelial cells and fusion and atrophy of foot processes occurred in the glomeruli of dogs with heartworm disease, but not in control dogs. The severity of tissue damage correlated with the degree of microfilaremia. There was no evidence that the glomerular lesions were caused by immune complexes deposited in the basement membranes; and the alterations of glomerular capillaries were probably caused by motile microfilariae in the capillaries.

Reabsorption of native and glycated albumin by renal proximal tubular epithelial cells

1996 ◽  
Vol 271 (2) ◽  
pp. F261-F268 ◽  
Author(s):  
M. Bendayan ◽  
I. Londono
Keyword(s):  
Epithelial Cells ◽  
Glomerular Filtration ◽  
Glycated Albumin ◽  
Basement Membranes ◽  
Endocytic Pathway ◽  
Tubular Epithelial Cells ◽  
Proximal Tubular Epithelial Cells ◽  
Proximal Tubular ◽  
Renal Proximal Tubular ◽  
Urinary Space

The proximal tubule epithelial handling of native and glycated albumin was evaluated by quantitative immunocytochemistry. Native bovine serum albumin (BSA) and its glycated form (gBSA), tagged to different haptens, were simultaneously injected in anesthetized mice and maintained in circulation for 10 or 60 min. Both albumins were localized within the capillary lumen, in glomerular and peritubular basement membranes, the urinary space, and cellular compartments of the proximal tubular epithelial cells. In these cells, both forms of albumin were concomitantly found within the same endocytic-lysosomal system. Morphometric evaluations have indicated higher proportions of gBSA in the urinary space, reflecting probably a significant glomerular filtration of this form of albumin combined to a lesser reabsorptive clearance. Indeed, higher proportions of native BSA were found in the endocytic compartment of the tubular epithelial cells, suggesting its preferential reabsorption. The present study thus supports a preferential glomerular filtration of gBSA with a facilitated filtration of native BSA in the presence of the glycated one. It also demonstrates the tubular reabsorption of BSA and gBSA through a common endocytic pathway, in which the native BSA is preferentially reabsorbed with respect to its glycated form.

scholarly journals Ultrastructural localization of the major components of the extracellular matrix in normal rat nerve.

1992 ◽  
Vol 40 (6) ◽  
pp. 859-868 ◽  
Author(s):  
P Lorimier ◽  
P Mezin ◽  
F Labat Moleur ◽  
N Pinel ◽  
S Peyrol ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Peripheral Nerve ◽  
Ultrastructural Localization ◽  
Basement Membranes ◽  
Type I ◽  
Immunoperoxidase Method ◽  
Adult Rat ◽  
Normal Rat ◽  
Rat Nerve ◽  
Fibronectin Type

In this study we determined the ultrastructural distribution of the various components of the extracellular matrix (laminin, fibronectin, Type I, III, and IV collagens) of the normal peripheral nerve in adult rat. The localization of these macromolecules was investigated in basement membranes as well as in different areas of epi-, peri-, and endoneurium, by use of a pre-embedding immunoperoxidase method.

scholarly journals UPTAKE OF EXOGENOUS HORSERADISH PEROXIDASE BY COATED VESICLES IN MOUSE NEUROMUSCULAR JUNCTIONS

1969 ◽  
Vol 17 (3) ◽  
pp. 161-170 ◽  
Author(s):  
SUMNER I. ZACKS ◽  
ATUSHI SAITO
Keyword(s):  
Skeletal Muscle ◽  
Horseradish Peroxidase ◽  
Synaptic Vesicles ◽  
Intramuscular Injection ◽  
Neuromuscular Junctions ◽  
Neuromuscular Function ◽  
Coated Vesicles ◽  
Intramuscular Nerves ◽  
Pinocytotic Vesicles ◽  
Intercellular Clefts

Following intramuscular injection of horseradish peroxidase into mouse skeletal muscle adjacent to areas of innervation, rapid uptake of the label was observed by coated but not synaptic vesicles. The tracer was also found in Schwann cells, external mesaxons and within peripheral myelin lamellae, but never in axoplasm of intramuscular nerves proximal to the injection site. The data suggest that the tracer is taken up by the coated vesicles and may be rapidly discharged into the synaptic clefts from which it is cleared by a combination of phagocytic activity and absorption via pinocytotic vesicles and intercellular clefts of adjacent capillaries or, more probably, enzyme activity is lost within the coated vesicles. The significance of these observations as related to neuromuscular function is discussed.

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.

Ultrastructural localization of lectin-binding sites in different basement membranes

1993 ◽  
Vol 25 (6) ◽  
pp. 464-468 ◽  
Author(s):  
Mehrdad Salamat ◽  
Werner G�tz ◽  
J�rgen Werner ◽  
Rainer Merken
Keyword(s):  
Binding Sites ◽  
Lectin Binding ◽  
Ultrastructural Localization ◽  
Basement Membranes ◽  
Lectin Binding Sites
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