Histochemical evaluation of glycosaminoglycan deposition in the skin.

Kupchella, Charles E.; Matsuoka, Lois Y.; Bryan, Bradley; Wortsman, Jacobo; J, Dietrich

doi:10.1177/32.10.6548236

Cited by 21 publications

(4 citation statements)

References 6 publications

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“…The various components of the preovulatory follicle are rich in glycoconjugates, which are often extracted during conventional processing for light and electron microscopy. Considerable success has been achieved by the inclusion of cetylpyridinium chloride (CPC) in the fixative [Pearse, 1980;Kupchella et al, 1984;Kop&y et al, 19841. Other methods to preserve proteoglycans include the use of cationic dyes such as Alcian blue [Goldberg et al, 19781, Ruthenium red [Luft, 1971;Myers et al, 19731, and Acridine orange [Lapis and Timar, 19801, which bind to anionic groups characteristic of acidic glycoconjugates.…”

Section: Introductionmentioning

confidence: 99%

Ultrastructural and histochemical changes in the murine zona pellucida during the final stages of oocyte maturation prior to ovulation

Kaufman¹,

Fowler²,

Barratt³

et al. 1989

Gamete Research

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The ultrastructural morphology of the mouse zona pellucida was studied in preovulatory follicles from the ovaries of immature mice treated with exogenous gonadotrophins. The ovaries were fixed in the presence of cetylpyridinium chloride, which precipitates carbohydrates, so that their loss during fixation and processing is substantially reduced. The semi-thin araldite sections obtained from osmicated material were viewed by conventional light microscopy, while the ultra-thin sections were examined by transmission electron microscopy. A parallel series of semi-thin sections of non-osmicated ovaries was deresined and subsequently stained with periodic acid Schiff (PAS). The morphological appearance of the zona pellucida in preovulatory oocytes changed during the final stages of oocyte maturation. A close correlation was observed between the ultrastructural appearance of the zona pellucida and that observed following PAS staining during the period studied. Real differences were observed in the location, density, and distribution of glycoconjugates within the zona pellucida during the final stages of oocyte maturation prior to and immediately following germinal vesicle breakdown. Similar changes in the zona were observed in adult females autopsied during proestrus and oestrus. The changes in the density and distribution of glycoconjugates are likely to have important consequences for fertilization by affecting sperm-zona binding and sperm-egg interactions.

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Section: Introductionmentioning

confidence: 99%

Ultrastructural and histochemical changes in the murine zona pellucida during the final stages of oocyte maturation prior to ovulation

Kaufman¹,

Fowler²,

Barratt³

et al. 1989

Gamete Research

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“…These staining procedures are normally used to detect glycosaminoglycans (also known as mucopoly saccharides). [27][28][29] The evanescence of the precipitate, that faded away with a slight agitation of the fixative, and the fact that it was only found when epe was used, and not with alde hydic fixatives alone, suggests that this is not a fibrin clot that can also be stained by the cationic dyes.…”

Section: Discussionmentioning

confidence: 99%

A gel of glycosaminoglycans lining the anterior and posterior chambers in man: Histochemical evidence at light and electron microscopy levels

Carreras

Caballero

Porcel

1992

Eye

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A thin mucous layer covers the ciliary body and iris epithelium and becomes thicker in the anterior chamber where it covers the anterior surface of the iris, the chamber angle and corneal endothelial surface. It is especially thick at the chamber angle, where it adopts a meniscus shape with the concavity towards the anterior chamber. Cetylpiridinium chloride along with glutaraldehyde was used to precipitate this layer of mucous substance lining the anterior and posterior chambers of the human eye. The staining of the precipitate by cationic dyes suggests that glycosaminoglycans are main components. The specific characterisation with anti-hyaluronic acid monoclonal antibody labelled with colloidal gold reveals the presence of hyaluronic acid in the precipitate. Long unbranched hyaluronic acid molecules may form the skeleton of a gel able to trap and hold virtually any other macromolecule suspended in the aqueous humour. A possible role of such a gel in the regulation of aqueous humour outflow is discussed.

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“…Other quaternary ammonium compounds (e.g., cetylpyridinium chloride) have also been shown to reduce the loss of PGs, but cetylpyridinium chloride interferes with the subsequent staining with cationic dyes, competing for the same anionic groups on PGs (Engfeldt & Hjertquist, 1968;Kupchella et al, 1984). Detailed quantitative data of the effects of cationic dyes on PG preservation during fixation and decalcification are still lacking.…”

Section: Introductionmentioning

confidence: 98%

Safranin O reduces loss of glycosaminoglycans from bovine articular cartilage during histological specimen preparation

et al. 1996

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The ability of Safranin O, added to fixation and decalcification solutions, to prevent the escape of glycosaminoglycans (GAGs) from small cartilage tissue blocks during histological processing of cartilage has been studied. GAGs in the fixatives and decalcifying solutions used and those remaining in the 1 mm3 cubes of cartilage were assayed biochemically. The quantity of GAGs remaining in the cartilage cubes were determined from Safranin O-stained sections using videomicroscopy or microspectrophotometry. A quantity (10.6%) of GAGs were lost during a conventional 4% buffered formaldehyde fixation (48 h) and a subsequent decalcification in 10% EDTA (12 days) at 4 degrees C. Roughly one-quarter of the total GAG loss occurred during the 48 h fixation, and three-quarters during the 12 days of decalcification. Inclusion of 4% formaldehyde in the decalcification fluid decreased the loss of GAGs to 6.2%. The presence of 0.5% Safranin O in the fixative reduced this loss to 3.4%. When 0.5% Safranin O was included in the fixative and 4% formaldehyde in the decalcification solution, Safranin O staining of the histological sections increased on average by 13.5%. After fixation in the presence of 0.5% Safranin O, there was no difference in the staining intensities when decalcification was carried out in the presence of either Safranin O or formaldehyde, or both. It took 24 h for Safranin O to penetrate into the deep zone of articular cartilage, warranting a fixation period of at least this long. In conclusion, the addition of Safranin O to the fixative and either Safranin O or formaldehyde in the following decalcification fluid, markedly reduces the loss of GAGs from small articular cartilage explants during histological processing. However, for immunohistochemical studies, Safranin O cannot be included in the processing solutions, because it may interfere.

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Histochemical evaluation of glycosaminoglycan deposition in the skin.

Cited by 21 publications

References 6 publications

Ultrastructural and histochemical changes in the murine zona pellucida during the final stages of oocyte maturation prior to ovulation

Ultrastructural and histochemical changes in the murine zona pellucida during the final stages of oocyte maturation prior to ovulation

A gel of glycosaminoglycans lining the anterior and posterior chambers in man: Histochemical evidence at light and electron microscopy levels

Safranin O reduces loss of glycosaminoglycans from bovine articular cartilage during histological specimen preparation

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