scholarly journals A histochemical method for the identification of 9-O-acyl sialic acid. An investigation of bovine submaxillary gland and intestinal mucins.

1978 ◽  
Vol 26 (3) ◽  
pp. 187-192 ◽  
Author(s):  
P E Reid ◽  
C F Culling ◽  
W L Dunn
Keyword(s):  
Sialic Acid ◽  
Potassium Hydroxide ◽  
Periodic Acid ◽  
Periodate Oxidation ◽  
Submaxillary Gland ◽  
Sialic Acids ◽  
Histochemical Method ◽  
Periodic Acid Schiff ◽  
Oxidation Step ◽  
Intestinal Mucins

Prolongation of the initial periodate oxidation step of the periodate-borohydride technique/potassium hydroxide/periodic acid-Schiff and periodic acid-Thionin Schiff/potassium hydroxide/periodic acid-Schiff sequences produced little or no change in the diagnostic staining for the potassium hydroxide/periodic acid-Schiff effect, exhibited by the colonic epithelial mucins of man and rat and the Brunner's gland mucin of rabbits. In contrast, there was a gradual, but clear decrease in the intensity of such staining of bovine submaxillary gland mucins. It was concluded that, in the intestinal mucins studied the potassium hydroxide/periodic acid-Schiff effect was due to sialic acids bearing O-acyl substitutents at positions C7 and/or C8 whereas in bovine submaxillary gland mucin the potassium hydroxide/periodic acid-Schiff effect is probably due, at least in part, to the presence of 9-O-acyl sialic acids. This investigation has led to the development of a technique which can be used to identify 9-O-acyl sialic acids.

scholarly journals A new histochemical method for the identification and visualization of both side chain acylated and nonacylated sialic acids.

1976 ◽  
Vol 24 (12) ◽  
pp. 1225-1230 ◽  
Author(s):  
C F Culling ◽  
P E Reid ◽  
W L Dunn
Keyword(s):  
Potassium Hydroxide ◽  
Periodic Acid ◽  
Sialic Acids ◽  
Histochemical Method ◽  
Side Chain ◽  
Parotid Glands ◽  
Periodic Acid Schiff ◽  
Mucin Histochemistry ◽  
Single Section ◽  
General Field

A new histochemical method is described for the differentiation of mucins that utilizes two different Schiff reagents and allows single section identification of side chain O-acylated, and nonacylated, sialic acids in contrasting colors. In the event of mucins containing only one type of sialic acid, it may allow their specific identification (e.g., C7 or C8 side chain O-acylated). It has been shown to be useful in the identification of some metastases from adenocarcinomas of colon (where the primary is potassium hydroxide/periodic acid-Schiff positive) and should prove of great value in the investigation of diseases of the gastrointestinal tract and particularly those of the colon. It should also be valuable in the general field of epithelial mucin histochemistry, particularly for those mucins of the salivary and parotid glands, etc.

scholarly journals THE HISTOCHEMICAL DEMONSTRATION OF O-ACYLATED SIALIC ACID IN GASTROINTESTINAL MUCINS THEIR ASSOCIATION WITH THE POTASSIUM HYDROXIDE-PERIODIC ACID-SCHIFF EFFECT

1974 ◽  
Vol 22 (8) ◽  
pp. 826-831 ◽  
Author(s):  
C. F. A. CULLING ◽  
P. E. REID ◽  
M. G. CLAY ◽  
W. L. DUNN
Keyword(s):  
Sialic Acid ◽  
Gastrointestinal Tract ◽  
Acid Hydrolysis ◽  
Potassium Hydroxide ◽  
Periodic Acid ◽  
Histochemical Demonstration ◽  
Sialic Acids ◽  
Side Chains ◽  
Periodic Acid Schiff ◽  
Alcoholic Potassium Hydroxide

O-Acylated sialic acids have been demonstrated histochemically in the epithelial mucins of the lower end of the gastrointestinal tract. The potassium hydroxide-periodic acid-Schiff effect has been shown to be due, in most areas, to the presence of these O-acyl esters on the polyhydroxy side chains of sialic acid. The esters have been shown to hinder the removal of the sialic acids by either neuraminidase or acid hydrolysis unless tissues were pretreated with alcoholic potassium hydroxide. It is suggested that substitution at the C4 position is responsible for their neuraminidase resistance.

scholarly journals A new histochemical method for detection of sialic acids using a physical development procedure.

1995 ◽  
Vol 43 (10) ◽  
pp. 1045-1051 ◽  
Author(s):  
T Ueda ◽  
O Fujimori ◽  
K Yamada
Keyword(s):  
Periodic Acid ◽  
Periodate Oxidation ◽  
Physical Development ◽  
Sialic Acids ◽  
New Method ◽  
Histochemical Method ◽  
Reaction Products ◽  
Specificity And Sensitivity ◽  
Development Procedure ◽  
Selective Periodate Oxidation

We have established an efficient histochemical method for demonstration of sialic acids in light microscopy. The method consists of a selective periodate oxidation-phenylhydrazine blockade and a thiocarbohydrazide-silver protein sequence followed by a physical development procedure. From the results obtained by the present experimental and control studies in tissue sections from a series of rat and mouse organs, such as stomach, duodenum, colon, liver, sublingual gland, lung, and kidney, the specificity and sensitivity of the method were sufficient. In the tissues tested, sialic acids were visualized as distinct brownish and blackish reaction products. Comparisons of the new method with the periodic acid-phenylhydrazine-Schiff (PA-P-S) or selective periodate oxidation-Schiff (PA*-S) method employed hitherto have confirmed that the new method reported here is higher in efficiency and visibility of reaction products than the latter two methods.

scholarly journals Apparent failure of 2-hydroxy-3-naphthoic acid hydrazide to block the periodic acid-Schiff reactivity of sialic acids. Implications for the PANFOPAS method.

1983 ◽  
Vol 31 (9) ◽  
pp. 1142-1144 ◽  
Author(s):  
P E Reid ◽  
C F Culling
Keyword(s):  
Sialic Acid ◽  
Periodic Acid ◽  
Acid Hydrazide ◽  
Sialic Acids ◽  
Side Chain ◽  
Substitution Pattern ◽  
Periodic Acid Schiff ◽  
Naphthoic Acid ◽  
Vicinal Diols ◽  
Anionic Groups

Histochemical studies of the mechanism of staining of the periodic acid/2,-hydroxy-3-naphthoic acid hydrazide/Fast Black B/saponification/periodic acid-Schiff (PANFOPAS) method indicate that while 2-hydroxy-3-naphthoic acid hydrazide blocks the periodate engendered Schiff reactivity of vicinal diols unassociated with anionic groups, it fails to block such reactivity in sialic acid residues. It is suggested, therefore, that the positive Schiff staining observed following application of the PANFOPAS method to colonic epithelial mucins may be due to sialic acids without side chain substituents (or which are substituted at C7 or C9) in addition to sialic acids substituted at C8 (or which are di- or trisubstituted). Consequently the PANFOPAS method cannot be used to study the side chain substitution pattern of the sialic acids of epithelial mucins.

scholarly journals THE HISTOCHEMICAL INTERPRETATION OF THE COMPLEX RESULTS OF METHYLATION UPON GASTROINTESTINAL TRACT MUCINS, WITH SPECIAL REFERENCE TO THE PERIODIC ACID-SCHIFF REACTIVITY

1974 ◽  
Vol 22 (10) ◽  
pp. 986-991 ◽  
Author(s):  
P. E. REID ◽  
C. F. A. CULLING ◽  
W. L. DUNN
Keyword(s):  
Sialic Acid ◽  
Gastrointestinal Tract ◽  
Special Reference ◽  
Periodic Acid ◽  
Separate Study ◽  
Sulfate Ester ◽  
Periodic Acid Schiff ◽  
Smith Degradation ◽  
Vicinal Diol ◽  
Schiff Reaction

The histochemical use of methylation has complex results; particularly in respect of the periodic acid-Schiff reaction, these are analyzed and discussed. Methods are described which allow the separate study of the following effects: (a) the removal of the KOH/periodic acid-Schiff effect; (b) removal of sialic acid from a potential vicinal diol; and (c) the removal of O-sulfate ester from a potential vicinal diol. The use of the Smith degradation technique, in addition to the above, also allows inferences to be drawn in respect of the structure of the mucins (glycoproteins) being investigated.

A physiological, biochemical and histological study of goose tracheal mucin and its secretion

1977 ◽  
Vol 279 (969) ◽  
pp. 513-540 ◽  
Keyword(s):  
Sialic Acid ◽  
Nerve Stimulation ◽  
Histological Study ◽  
Periodic Acid ◽  
Gel Filtration ◽  
Goblet Cells ◽  
Mucin Secretion ◽  
Electron Microscopic ◽  
Periodic Acid Schiff ◽  
Tracheal Mucus

Tracheal mucin secretion has been measured from a segment of trachea, isolated in situ , in anaesthetized geese by a method that involves radioactive labelling of tracheal mucus glycoproteins (Gallagher et al. 1975). Goose tracheal mucus comes entirely from goblet cells, since the goose trachea does not contain submucosal mucous or serous glands, and this method has been used to investigate the nervous and pharmacological control of the mucin secretion from these epithelial goblet cells. The mucins secreted have been collected, fractionated, and chemically analysed. Intracellular mucin has been examined histochemically, and the results of electron microscopic observations of epithelial cells and nerves are presented. Acetylcholine increased tracheal mucin secretion, and this effect was completely blocked by atropine. Neither α- nor β-stimulant sympathomimetic amines affected tracheal mucin secretion. Stimulation of the peripheral cut ends of the descending oesophageal nerves increased tracheal mucin secretion and the majority of this response, approximately three-quarters, appeared to be cholinergic since this proportion was blocked by atropine. The mediator for the atropine-resistant part of the response is not known, but it appears not to be a β-adrenoreceptor stimulant since the response to nerve stimulation was unaffected by propranolol given at 34 μm intrasegmentally. Other possibilities are discussed. Atropine itself decreased the resting level of tracheal mucin secretion. The local anaesthetic, lignocaine, increased tracheal mucin secretion, while at the same time blocking the responses to acetylcholine and descending oesophageal nerve stimulation. The implications of this are discussed. The electrophoretic, gel filtration and ion-exchange properties of goose tracheal mucins showed that they represented high molecular mass, negatively charged glycoproteins which could be labelled biosynthetically with [ 35 S]sulphate, [ 3 H]- and [ 14 C]glucose. These mucins could be stained with Alcian blue or periodic acid Schiff reagent. The carbohydrate composition was unusual for an epithelial glycoprotein in that fucose was absent and mannose was present in small quantities. The monosaccharides present in larger quantity were galactose, N -acetylglucosamine, N -acetylgalactosamine and sialic acid. Histochemical analysis of tissue sections of gosling tracheas demonstrated that nearly all of the glycoprotein in epithelial goblet cells contained both sialic acid and sulphate residues. Sialated mucin was present also, but to a lesser extent, and many cells contained a mixture of sialated and sulphated mucins. The adult goose trachea had a high proportion of sialated glycoprotein. Electron microscopy showed a range of epithelial cell types and intra-epithelial nerves also. Many of the nerves had neurosecretory vesicles suggestive of motor function and some were near to goblet cells.

scholarly journals Detection in milk of a 60,000 Mr mouse and a 68,000 Mr human phosphorylated glycoprotein by histochemical analysis on polyacrylamide gels.

1983 ◽  
Vol 31 (6) ◽  
pp. 709-716 ◽  
Author(s):  
M R Green
Keyword(s):  
Molecular Weight ◽  
Sialic Acid ◽  
Periodic Acid ◽  
Sodium Dodecyl ◽  
Apparent Molecular Weight ◽  
Blue Staining ◽  
Polyacrylamide Gels ◽  
Periodic Acid Schiff ◽  
Coomassie Blue Staining ◽  
Coomassie Blue

Proteins in colostrum and skimmed milk from humans and mice were separated by electrophoresis on polyacrylamide gels and stained with Coomassie blue (CB), Ethyl-Stains-all (ESA), and periodic acid-Schiff (PAS) to investigate changes that may occur in milks throughout lactation. In mouse colostrum but not in mature mouse milk, a PAS-positive protein of apparent molecular weight of 60,000 stained prominently blue with ESA. A protein in human milk with a molecular weight of 68,000 stained similarly but was present throughout lactation. The intensity of blue staining of these minor proteins in milk approached that obtained with casein phosphoproteins. The metachromatic dye ESA stains phosphoproteins and sialic acid-rich glycoproteins blue to blue-green. Removal of phosphorus from the former and sialic acid from the latter results in those proteins staining red with ESA. The intensity of blue staining of the 60,000 and 68,000 Mr proteins was diminished but not lost following treatment with phosphatase. It was eliminated following neuraminidase digestion of the mouse protein and mild acid hydrolysis of the human protein. Coomassie blue staining of the proteins was not affected by these procedures. Following electrophoresis of milk and milk fractions in a non-sodium dodecyl sulfate-containing system, the proteins were identified by their characteristic staining properties with ESA and isolated.

scholarly journals Histochemical reactivity of peanut lectin-horseradish peroxidase conjugate.

1980 ◽  
Vol 28 (9) ◽  
pp. 979-990 ◽  
Author(s):  
P J Stoward ◽  
S S Spicer ◽  
R L Miller
Keyword(s):  
Sialic Acid ◽  
Horseradish Peroxidase ◽  
Periodic Acid ◽  
Apical Cell ◽  
Goblet Cells ◽  
Surface Epithelium ◽  
Renal Tubules ◽  
Peanut Lectin ◽  
Periodic Acid Schiff ◽  
Terminal Galactose

A peanut lectin-horseradish peroxidase (PL-HRP) conjugate has been applied to histochemical staining of paraffin sections of various mouse organs. The PL-HRP conjugate has selectively reacted with secretory bodies, the Golgi zone, and the apical cell surface in various cell types. Some positive sites, including lingual and tracheal serous glands, Brunner's glands, and the brush border of the proximal straight nephron, contained periodic acid-Schiff (PAS)-positive glycoconjugate with no affinity for basic reagents. The stored secretion in these sites was interpreted as containing neutral glycoprotein with terminal galactose residues which could, in part at least, account for the PAS reactivity. Duodenal goblet cells, which exhibited basophilia attributable to sulfate esters, also bound PL-HRP. As the binding was affected by prior sialidase digestion, the secretory glycoprotein in the duodenal goblet cells was judged to contain oligosaccharides with sulfate esters and terminal galactose uncapped by sialic acid. All sites known from their basophilia to form sialomucin failed to stain with the PL-HRP conjugate, but consistently gained reactivity following sialidase digestion and were inferred, therefore, to possess glycoproteins with oligosaccharide side chains containing subterminal galactose and terminal sialic acid. Lingual mucous glands, known to secrete a mucosubstance with basophilic properties indicative of the presence of sulfate esters but not sialic acid, stained with PL-HRP only after sialidase digestion and, accordingly, were reinterpreted as containing both sulfate esters and terminal galactose-sialic acid dimers. Staining of gastric surface epithelium demonstrated a srongly PAS-reactive neutral glycoprotein, and that of goblet cells in the cecum disclosed PAS-positive sulfated glycoprotein. The latter two sites lacked PL-HRP affinity without or with prior sialidase treatment and apparently possessed neither terminal galactose residues nor galactose-sialic acid dimers. PL-HRP affinity was observed exclusively in the Golgi cisternae of some epithelial cells, thus indicating that galactose occurs transiently as a terminal residue in this site. A few histologic sites, such as pancreatic and gastric zymogen cells and renal tubules, were devoid of both PAS reactivity and basophilia indicative of the presence of complex carbohydrate but stained strongly with the PL-HRP conjugate by means which are not understood. Galactose in the PL-HRP solution blocked or reversed the PL-HRP binding in most of the structures with an affinity for the conjugate, supporting the conclusions that the reagent is specific for galactosyl residues.

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