scholarly journals Detection of glomerular anionic sites in post-embedded ultra-thin sections using cationic colloidal gold.

1991 ◽  
Vol 39 (7) ◽  
pp. 965-972 ◽  
Author(s):  
N P Goode ◽  
M Shires ◽  
D M Crellin ◽  
A M Davison
Keyword(s):  
Hyaluronic Acid ◽  
Chondroitin Sulfate ◽  
Colloidal Gold ◽  
Endothelial Glycocalyx ◽  
Computer Assisted ◽  
Nuclear Staining ◽  
Anionic Sites ◽  
Thin Sections ◽  
Cationic Colloidal Gold ◽  
Influence Of Ph

We detected glomerular anionic sites in fixed, LR Gold-embedded ultra-thin tissue sections using cationic colloidal gold. Manual and computer-assisted quantitation were compared, and the influence of pH and glycosaminoglycan-degrading enzymes on site expression was examined. Both quantitation methods produced similar results. Alteration of pH within a narrow range (pH 2.5-3.0) markedly affected the staining pattern. At pH 2.5, epithelial and endothelial glycocalyx and regular sites restricted to the lamina rara externa were stained. At pH 3.0 and above, glycocalyx was unstained but intracellular and nuclear staining was present; glomerular basement membrane (GBM) and mesangial matrix sites were abundant. After chondroitinase ABC or hyaluronidase digestion, GBM staining was eliminated at pH 2.0 and reduced at pH 7.0 (p less than 0.001), suggesting that degraded sites are associated with chondroitin sulfate or hyaluronic acid. By contrast, prolonged heparitinase I digestion was ineffective at either pH. Digestion of purified substrates revealed crossreactivity of heparitinase towards chondroitin sulfate and of chondroitinase towards hyaluronic acid. Since tissue sites were reduced by chondroitinase but not heparitinase, we suggest that degradation is due to hyaluronidase activity of chondroitinase and the anionic sites are associated with hyaluronic acid. However, the influence of pH indicates that lamina rara externa sites are structurally distinct from other GBM anionic sites.

Visualization of the anionic sites in the cell wall of apple fruit using a cationic colloidal gold probe

1993 ◽  
Vol 51 ◽  
pp. 320-321
Author(s):  
Stéphane Roy ◽  
William S. Conway ◽  
Alley E. Watada ◽  
Christopher D. Pooley ◽  
William P. Wergin
Keyword(s):  
Cell Wall ◽  
Colloidal Gold ◽  
Apple Fruit ◽  
Gold Complex ◽  
Anionic Sites ◽  
Wall Strength ◽  
Anionic Binding Sites ◽  
Cationic Colloidal Gold ◽  
Softening Process ◽  
Gold Probe

The ripening of fleshy fruits involves a softening process that consists of biochemical changes in the cell wall and leads to cell separation. Calcium is an important constituent of the cell wall and plays roles in maintaining the firmness of fruit and in reducing postharvest decay. The modification of cell wall strength is believed to be influenced by calcium that interacts with acidic pectic polymers to form crossbridges. This study examined how the frequency and distribution of anionic binding sites in the cell walls of apple fruit were influenced by calcium infiltration.Mature “Golden Delicious” apple fruits were pressure infiltrated with either H2O or a 4% solution of CaCl2 and the pericarp was sampled and processed according to standard procedures. Cationic poly-Llysine colloidal gold complex was used in a one-step procedure to visualize anionic sites in muro. Observations were performed with light microscopy, following silver intensification, and with transmission electron microscopy.

scholarly journals Demonstration of anionic sites in human eccrine and apocrine sweat glands in post-embedded ultra-thin sections with cationic colloidal gold: effect of enzyme digestion on these anionic sites.

1993 ◽  
Vol 41 (8) ◽  
pp. 1197-1207 ◽  
Author(s):  
K Saga ◽  
M Takahashi
Keyword(s):  
Colloidal Gold ◽  
Dermatan Sulfate ◽  
Sweat Glands ◽  
Secretory Cells ◽  
Anionic Sites ◽  
Chondroitinase Abc ◽  
Dark Cell ◽  
Luminal Membrane ◽  
Cationic Colloidal Gold ◽  
Eccrine Sweat Glands

We localized anionic sites ultrastructurally in human eccrine and apocrine sweat glands with a poly-L-lysine-gold complex (cationic colloidal gold). Anionic sites were labeled by incubating Lowicryl K4M-embedded sections on droplets of cationic colloidal gold. In eccrine sweat glands, colloidal gold particles were restricted to the basolateral membrane of the secretory cells at low pH, whereas the luminal membrane did not react with the gold particles. Chondroitinase ABC digested these anionic sites. This indicates that chondroitin sulfate and/or dermatan sulfate constitutes anionic sites in the basal labyrinth of eccrine sweat glands. In apocrine sweat glands, the luminal membrane of the secretory cells showed strong reaction at low pH, whereas the contraluminal membrane did not show any reaction. Neuraminidase completely digested these anionic sites, which indicated that the anionic charge of the apocrine lumen was due to sialic acid. Differences in distribution and susceptibility to enzymes of anionic sites in cell membranes between eccrine and apocrine sweat glands may reflect functional differences between these glands. Dark cell granules in eccrine secretory cells were negative for the anionic sites when sections were labeled without any pre-treatment. However, pre-incubation of the grids on EGTA or deionized water unmasked the anionic sites on the dark cell granules. The positive staining after EGTA treatment was greatly decreased by reincubation with CaCl2. These results suggested that Ca blocked anionic sites in dark cell granules. Exposed anionic sites were digested with chondroitinase ABC. This indicated that chondroitinase ABC and/or dermatan sulfate composed the anionic sites in dark cell granules.

3-D organization of fibrinogen receptors using computer reconstruction and whole-mount microscopy

1988 ◽  
Vol 46 ◽  
pp. 30-31
Author(s):  
E. T. O'Toole ◽  
R. R. Hantgan ◽  
J. C. Lewis
Keyword(s):  
Whole Blood ◽  
Colloidal Gold ◽  
Blood Platelets ◽  
Cell Contact ◽  
Computer Assisted ◽  
Adherent Cells ◽  
Differential Centrifugation ◽  
Computer Reconstruction ◽  
Sds Page ◽  
Whole Mount

Thrombocytes (TC), the avian equivalent of blood platelets, support hemostasis by aggregating at sites of injury. Studies in our lab suggested that fibrinogen (fib) is a requisite cofactor for TC aggregation but operates by an undefined mechanism. To study the interaction of fib with TC and to identify fib receptors on cells, fib was purified from pigeon plasma, conjugated to colloidal gold and used both to facilitate aggregation and as a receptor probe. Described is the application of computer assisted reconstruction and stereo whole mount microscopy to visualize the 3-D organization of fib receptors at sites of cell contact in TC aggregates and on adherent cells.Pigeon TC were obtained from citrated whole blood by differential centrifugation, washed with Ca++ free Hank's balanced salts containing 0.3% EDTA (pH 6.5) and resuspended in Ca++ free Hank's. Pigeon fib was isolated by precipitation with PEG-1000 and the purity assessed by SDS-PAGE. Fib was conjugated to 25nm colloidal gold by vortexing and the conjugates used as the ligand to identify fib receptors.

Glycosaminoglycans from growing antlers of wapiti (Cervus elaphus)

1997 ◽  
Vol 77 (4) ◽  
pp. 715-721 ◽  
Author(s):  
H. H. Sunwoo ◽  
L. Y. M. Sim ◽  
T. Nakano ◽  
R. J. Hudson ◽  
J. S. Sim
Keyword(s):  
Hyaluronic Acid ◽  
Chondroitin Sulfate ◽  
Cervus Elaphus ◽  
Molecular Size ◽  
Enzymatic Digestion ◽  
Gel Chromatography ◽  
Cellulose Acetate Electrophoresis ◽  
Cartilaginous Tissues ◽  
Chondroitin Sulfates ◽  
Velvet Antler

The emerging wapiti industry in North America is based largely on markets for velvet antlers which are used in oriental medicine. Despite the economic opportunity, enthusiasm has been dampened by incomplete understanding of the chemical and pharmacological properties of velvet antler. This study characterizes polysaccharide constituents of glycosaminoglycans in growing antler of wapiti (Cervus elaphus). Glycosaminoglycans were isolated from four sections (tip, upper, middle and base) of growing antlers, and were studied using cellulose acetate electrophoresis, gel electrophoresis, enzymatic digestion and gel chromatography. The tip and upper sections of the antler which are rich in cartilaginous tissues contained chondroitin sulfate as a major glycosaminoglycan with small amounts of hyaluronic acid. In the middle and base sections containing bone and bone marrow, chondroitin sulfate was also a major glycosaminoglycan with small amounts of hyaluronic acid and chondroitinase-ACI resistant materials. More than half of chondroitin sulfate from the middle and base sections had larger molecular size than did the chondroitin sulfates from the tip and upper sections. Key words: Glycosaminoglycans, chondroitin sulfate, antler, wapiti

scholarly journals Post-embedding colloidal gold immunolocalization of laminin to the lamina rara interna, lamina densa, and lamina rara externa of renal glomerular basement membranes.

1986 ◽  
Vol 34 (7) ◽  
pp. 847-853 ◽  
Author(s):  
D R Abrahamson
Keyword(s):  
Colloidal Gold ◽  
Basement Membranes ◽  
Lamina Densa ◽  
Thin Sections ◽  
Gold Particles ◽  
Rabbit Igg ◽  
Gold Immunolocalization ◽  
Lowicryl K4m

Ultrastructural distribution of laminin within renal glomerular (GBM) and tubular basement membranes (TBM) was investigated using post-embedding immunolocalization with colloidal gold. Rat kidneys were fixed with 4% formaldehyde and embedded at 4 degrees C in Lowicryl K4M medium. Thin sections were then sequentially treated with affinity-purified rabbit anti-laminin IgG and anti-rabbit IgG conjugated to 10 nm diameter colloidal gold. Gold bound specifically to the GBM and TBM with particle densities of 690/micron2 and 731/micron2, respectively. In the GBM, the number of gold particles bound/micron2 of lamina densa greater than lamina rara externa greater than lamina rara interna. Closely similar binding patterns were found when kidneys were fixed with 0.5% glutaraldehyde plus 3% formaldehyde and embedded at 60 degrees C in L.R. White resin, but slightly less gold bound to sections overall than that seen with formaldehyde alone and Lowicryl. Taken together, these results illustrate that anti-laminin IgG, whether applied to fixed sections in vitro or introduced in vivo, bound to the lamina rara interna, lamina densa, and lamina rara externa of the GBM and throughout the TBM.

PO-01-112 Hyaluronic acid and chondroitin sulfate, alone or in combination, efficiently counteract induced bladder cells damage and inflammation

2019 ◽  
Vol 16 (5) ◽  
pp. S75-S76
Author(s):  
C. Schiraldi ◽  
A. Stellavato ◽  
A. V. Adriana Pirozzi ◽  
P. Diana ◽  
G. Donnarumma ◽  
...  
Keyword(s):  
Hyaluronic Acid ◽  
Chondroitin Sulfate ◽  
Bladder Cells
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