thiocarbohydrazide and metaperiodate

Oxidation of araldite-embedded liver sections by 1% w/v aqueous H5IO6 for 15 min and a 5-min reaction of carbonyls with 1% w/v thiocarbohydrazide in 10% v/v acetic acid was employed for subsequent staining of glycogen with silver-proteinate (S-P). The network of branching intracellular glycogen aggregates was revealed by 15-min staining with S-P, whereas 24 hr incubation in S-P was necessary to enhance the contrast of glycogen inclusions. We conclude that the proposed modification of glycogen staining readily affords the means for its localization at a desired level of contrast and resolution.

Topics: Animals; Glycogen; Histocytochemistry; Hydrazines; Liver; Microscopy, Electron; Periodic Acid; Rats; Rats, Inbred Strains; Silver Proteins; Staining and Labeling

Complex carbohydrates in secretory granules and at the apical cell surface of mouse gastric mucoid cells were studied during embryogenesis and in the early postnatal period by various cytochemical methods; the periodic acid-thiocarbohydrazide-silver proteinate (PA-TCH-SP) and tannic acid-uranyl acetate (TA-UA) procedures made neutral mucosubstances (NMS) visible, whereas the hexose residues of glycoconjugates were identified using WGA-, RCA II- and ConA-ferritin. The glycocalyx was stained with ruthenium red (RR). During differentiation of the embryonic mucoid cells the number of secretory granules increased in parallel to the increase in their carbohydrate component. NMS-stainable parts in secretory granules also had binding sites for the conjugates RCA II- and WGA-ferritin, but the binding of ConA could not be identified. The increasing quantity of NMS in secretory granules was correlated with the increased amount of PA-TCH-SP and TA-UA positive substances in the apical glycocalyx only in 14- and 18-day-old embryos. The observed uniform affinity for RR and lectin conjugates in all analysed developmental stages remains to be explained.

Topics: Animals; Animals, Newborn; Cell Membrane; Concanavalin A; Cytoplasmic Granules; Embryo, Mammalian; Ferritins; Gastric Mucosa; Glycosaminoglycans; Histocytochemistry; Horseradish Peroxidase; Hydrazines; Hydrolyzable Tannins; Lectins; Mice; Organometallic Compounds; Periodic Acid; Plant Lectins; Ruthenium Red; Silver Proteins; Staining and Labeling; Uranium; Wheat Germ Agglutinins

Glycoconjugates were localized by light microscopy with lectin-peroxidase conjugates and by electron microscopy with the periodic acid-thiocarbohydrazide-silver proteinate (PA-TCH-SP) sequence in immunocytochemically or morphologically identified cell types in rat pituitary. Lectin histochemistry demonstrated sialic acid and glycoconjugates with N-glycosidically linked oligosaccharides in gonadotrophs, thyrotrophs, and corticotrophs. Galactose penultimate to sialic acid was observed mostly in gonadotrophs. The terminal galactose-N-acetylgalactosamine disaccharide was detected in a few gonadotrophs and in a moderate number of mammotrophs. Fucose was localized in only corticotrophs with two fucose-binding lectins and in thyrotrophs with another. Several different monosaccharides were seen in glycoconjugates in melanotrophs and in Herring bodies. Melanotrophs displayed heterogeneous staining with fucose-binding lectins. A small number of nonsecretory cells were also visualized in the pars distalis by virtue of their glycogen content. PA-TCH-SP staining revealed complex carbohydrates in secretory granules and some Golgi cisternae in all types of hormone-producing cells in the pars distalis except for the somatotrophs. Melanotrophs of pars intermedia exhibited stained secretory granules and irregular dense bodies containing a stained meshwork. Corticotrophs of the pars distalis lacked the latter bodies, although they form the same glycoprotein precursor hormone as melanotrophs. Lectin conjugates and the PA-TCH-SP sequence stained some groups of secretion granules in Herring bodies, possibly representing vasopressin-containing granules as well as other cell types in the pars nervosa.

Topics: Animals; Carbohydrates; Cosyntropin; Follicle Stimulating Hormone; Glycogen; Glycoproteins; Histocytochemistry; Hydrazines; Immunoenzyme Techniques; Indicators and Reagents; Lectins; Luteinizing Hormone; Male; Melanins; Microscopy, Electron; Oligosaccharides; Periodic Acid; Pituitary Gland; Rats; Rats, Inbred Strains; Silver Proteins; Thyrotropin; Tissue Distribution

An ultrastructural histochemical study was carried out on submandibular glands of rabbits. Stainings were performed with dialysed iron (DI), high iron diamine (HID), tannic acid uranyl acetate (TA-U), tannic acid-ferric chloride (TA-F) sequence, and periodate-thiocarbohydrazide-silver proteinate (PA-TCH-SP) method. It was demonstrated that neutral glycoproteins are present in the cells with dark granules of the preterminal tracts, and that neutral and acid glycoproteins are present in the cells with light granules of the terminal tracts. Result are discussed and compared to other previously obtained histochemical and biochemical data.

Topics: Animals; Diamines; Female; Glycoproteins; Histocytochemistry; Hydrazines; Hydrolyzable Tannins; Iron; Periodic Acid; Rabbits; Silver; Staining and Labeling; Submandibular Gland