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Query: UNIPROT:P02794 (
ferritin
)
17,525
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
Dissociation of adult cardiac myocytes by collagenase perfusion techniques requires separation of the junctional contacts that link the cells physically, electrically and metabolically in the intact heart. Gap junctions, one of three types of intercellular junction present at the cardiac intercalated disc, are not split into their component membranes when myocytes are dissociated; they are ripped from the plasma membrane of one cell, to be retained by its neighbour. Partitioning of junctions in this way might be expected to constitute a serious threat to the ionic integrity of dissociated myocytes, but in practice, high yields of functionally intact cells, suitable for experimental studies, are routinely obtained. To explain this apparent paradox, repair mechanisms, operating to seal the membrane lesions caused by gap junction tearing, have been hypothesized, but evidence for their existence has previously been lacking. Using freeze-fracture electron microscopy, the present study identifies repair sites as smooth membrane domains that are continuous with the neighbouring plasma membrane, thus forming intact seals. That these structures are not chemically-induced artefacts is demonstrated by their presence in myocytes that were frozen directly from the living state. Subsarcolemmal vesicle clusters, detected in thin sections and freeze-fracture replicas, are associated with the smooth sealing domains. These structures may represent either rounded-up fragments of mechanically disrupted membrane or structures concerned with the synthesis of new lipid. From their freeze-fracture morphology, the sealing domains appear to be lipid-rich and protein-poor. Cytochemical studies using
Ruthenium
Red, cationized
ferritin
and lectins show in addition that they have a lower content of negatively-charged membrane components than the neighbouring plasma membrane, and that the carbohydrate residues normally associated with plasma membrane glycolipids and glycoproteins are absent.
...
PMID:Integrity of the dissociated adult cardiac myocyte: gap junction tearing and the mechanism of plasma membrane resealing. 235 53
1. The hydraulic conductance of the synovial lining of a rabbit knee increases greatly when intra-articular pressure (IAP) is raised above approximately 9 cmH2O (yield point). To investigate the cause, synovium was fixed in situ by perfusion at controlled IAP and prepared for transmission electron microscopy. Micrographs of synovium fixed below yield pressure (atmospheric pressure and 5 cmH2O IAP, ten joints) and above it (25 cmH2O IAP, five joints) were analysed by morphometry. 2. The discontinuous cellular lining consisted of fibroblast-like cells (67%) and macrophage-like cells (33%) separated by interstitium-filled gaps. Interstitium formed 26-36% of the surface below yield pressure. Depending on sample site the surface gaps averaged 1.9 +/- 0.2 to 2.4 +/- 0.2 microns wide below yield pressure (mean +/- S.E.M. throughout). Above yield pressure the mean gap width increased by 42-64% (P less than 0.05, analysis of variance). 3. The qualitative and quantitative composition of the lining varied with distance below the surface. In a plane 5 microns deep, the intercellular distances and interstitial area fraction were almost double those at the surface. Classic periodic collagen fibrils (diameter 50 +/- 3 nm) abounded at 5 microns depth whereas the surface interstitium was richer in
Ruthenium
Red-staining microfibrils (diameter 9.3 +/- 0.7 nm) associated with 93 nm period fibrous long-spacing bundles. 4. Averaging over all the tissue between the surface and the 5 microns deep plane, the mean interstitial volume fraction was 0.61 +/- 0.05 at 5 cmH2O and 0.67 +/- 0.02 at 25 cmH2O (n.s.). 5. Capillary fenestrae (8.5 +/- 1.1 per fenestrated profile) and intercellular junctions were unaltered at high IAP. The tortuosity of the capillary-to-joint cavity path was 1.50 +/- 0.01 below yield pressure and 1.86 +/- 0.24 at 25 cmH2O (n.s.). 6. Intra-articular tracers (ferrocyanide,
ferritin
and glycogen) permeated synovial interstitium without evidence of preferential pathways. Ferrocyanide delineated the capillary intercellular junction as a permeable channel. Ferritin and glycogen were phagocytosed by the macrophages. 7. In suprapatellar areolar synovium, the most extensive and most altered tissue, the ratio of interstitial area to path length increased maximally 4.1 times between 5 and 25 cmH2O IAP. This represents a substantial contribution to the physiologically estimated rise in interstitial conductance (14 x) but does not wholly explain it.
...
PMID:Ultrastructure of transport pathways in stressed synovium of the knee in anaesthetized rabbits. 262 39
A method using low concentrations of formaldehyde and dithiothreitol was applied to obtain 'right-side out' luminal plasmalemma-derived vesicles from bovine aortic endothelial cells (EC) in culture, and from human umbilical vein and bovine or porcine aortas perfused ex vivo with the vesiculation solution. Vesicle formation and shedding were examined by phase-contrast microscopy and by transmission (TEM) and scanning electron microscopy (SEM). Vesicles showed the characteristic trilaminar pattern of the unit membrane and did not contain cellular organelles. As detected in freeze-fracture preparations, vesicle membrane displayed intramembrane particles and filipin-detectable cholesterol. Like EC plasmalemma, vesicle surface was heavily stained by
Ruthenium
Red and bound under a normal pattern cationized
ferritin
and
ferritin
hydrazide. As indicated by lectin agglutination assays and by ultrastructural cytochemistry, vesicles maintained on their ectodomains glycoconjugates bearing monosaccharides such as N-acetyl-neuraminic acid, beta-N-acetylglucosamine and beta-D-galactose, and expressed 5'-nucleotidase activity. The electrophoretic profiles of externally disposed 125I-labelled polypeptides of vesicles were found to be similar to those of intact EC. Chemically-induced vesiculation appears as a suitable method to obtain EC plasmalemma for studying its composition and functions in various vascular beds.
...
PMID:Endothelial cell plasma membrane obtained by chemically induced vesiculation. 359 39
The surface coating of the pneumonocytes in the lungs of five human neonates was studied by conventional electron microscopy and by cytochemical techniques. These techniques included staining with colloidal iron oxide, cationic
ferritin
, Ruthenium red,
Ruthenium
hexamine trichloride, high iron diamine and the periodic acid-thiocarbohydrazide-silver proteinate sequence. The results of the study indicate that the surface-bound coat of the pneumonocytes in human neonatal lung is a periodate-reactive sialomucin.
...
PMID:The surface coating of the pneumonocytes in human neonatal lung. 620 80