scholarly journals CORRELATED LIGHT AND ELECTRON MICROSCOPE OBSERVATIONS ON GLYCOPROTEIN-CONTAINING GLOBULES IN THE FOLLICULAR CELLS OF THE THYROID GLAND OF THE RAT

1965 ◽  
Vol 13 (4) ◽  
pp. 286-295 ◽  
Author(s):  
HUBERTA E. VAN HEYNINGEN
Keyword(s):  
Electron Microscope ◽  
Thyroid Gland ◽  
Toluidine Blue ◽  
Fine Particles ◽  
Periodic Acid ◽  
Follicular Cells ◽  
Periodic Acid Schiff ◽  
Intense Staining ◽  
Thin Sections ◽  
Rat Thyroid

Two carbohydrate staining techniques were applied to sections of rat thyroid gland: periodic acid-silver methenamine to thin sections for electron microscopy, and periodic acid-Schiff to thick sections for light microscopy. The latter were compared with adjacent thin sections for identificatoin with the electron microscope. Two types of globules in thyroid follicular cells stained with both methods. Globules of the first type are relatively large (usually 0.5 to 3 µ) with electron opacity very similar to that of follicular colloid; when stained with toluidine blue they have the same gray shade as follicular colloid. These similarities suggest that their periodic acid reactivity is due to the same glycoprotein as that of follicular colloid, namely thyroglobulin, and that the origin of these "intracellular colloid droplets" is the colloid in the lumen. The second type comprises medium-sized (usually 0.1 to 1 µ) fairly electron opaque globules having fine particles (~70 Å) dispersed in their matrices and sometimes containing membrane fragments or other irregularities; when stained with toluidine blue these globules stand out dark blue. Although their periodic acid reactivity indicates that these globules also contain glycoprotein, their ultrastructure and staining characteristics suggest that their composition differs from colloid. It is possible that they represent enzyme or zymogen-containing granules. A third type of globule, which on account of its intense staining in some periodic acid silver methenamine preparations could perhaps also be periodic acid reactive, concerns small (usually 0.02 to 0.2 µ) globules, mainly accumulated beneath the apical border of the cell. These globules, known as "apical vesicles," are believed to contain the not-yet-iodinated precursor of thyroglobulin.

scholarly journals Immunocytochemical localization of cathepsins B, H, L, and T4 in follicular cells of rat thyroid gland.

1989 ◽  
Vol 37 (5) ◽  
pp. 691-696 ◽  
Author(s):  
Y Uchiyama ◽  
T Watanabe ◽  
M Watanabe ◽  
Y Ishii ◽  
H Matsuba ◽  
...  
Keyword(s):  
Thyroid Gland ◽  
Thyroid Tissue ◽  
Double Immunostaining ◽  
Follicular Cells ◽  
Thin Sections ◽  
Dense Granules ◽  
Dense Bodies ◽  
Rat Thyroid ◽  
Monospecific Antibodies ◽  
Rat Thyroid Gland

To localize cathepsins B, H, and L in follicular cells of rat thyroid gland, we applied immunocytochemistry to the thyroid tissue using their respective monospecific antibodies. On serial semi-thin sections, cathepsins B, H, and L were localized in granules of various sizes located throughout the cytoplasm, whereas T4 was detected in larger granules located in the apical and supranuclear regions. By electron microscopy, cathepsins B, H, and L were localized in large less-dense granules (so-called colloid droplets) and in dense bodies of various sizes, whereas T4 was localized more intensely in large less-dense granules than in smaller dense bodies. By double immunostaining using an immunogold method, cathepsins H and B or L were co-localized in the same cytoplasmic granules. Moreover, immunoblotting demonstrated that proteins similar to cathepsins B, H, and L in the liver are present in the thyroid gland. These results suggest that cathepsins B, H, and L participate not only in degradation of thyroglobulin but in maturation of thyroid hormones, although it remains unknown whether all of them participate in the maturation process.

scholarly journals Immunocytochemical localization of glycoprotein hormones in the rat anterior pituitary. A light and electron microscope study using antisera against rat bet subunits: a comparison between preembedding and postembeeding methods.

1980 ◽  
Vol 28 (2) ◽  
pp. 101-114 ◽  
Author(s):  
C Tougard ◽  
R Picart ◽  
A Tixier-Vidal
Keyword(s):  
Electron Microscope ◽  
Binding Sites ◽  
Secretory Granules ◽  
Periodic Acid ◽  
Light And Electron Microscopy ◽  
Thyroid Stimulating Hormone ◽  
Periodic Acid Schiff ◽  
Thyrotropic Cells ◽  
Rat Thyroid ◽  
Stimulating Hormone

The binding sites of antisera (anti) to the beta (beta) subunits of rat follicle-stimulating hormone (rFSH), rat luteinizing hormone (rLH), and rat thyroid-stimulating hormone (rTSH) have been localized in rat anterior pituitaries by immunocytochemistry using light and electron microscopy. With the light microscope, LHbeta and FSHbeta were found in the same cells, which were violet after the alcian blue-periodic acid Schiff (AB-PAS) staining. TSHbeta was found in polygonal or stellate cells that were blue after AB-PAS. With the electron microscope, the thyrotropic cells contained very small secretory granules. LHbeta and FSHbeta were found in various types of cells (types A and B and their intermediate forms), which had previously been identified as gonadotropic cells. On serial ultrathin sections using the postembedding method the same cells and even some granules inside these cells were stained by both anti-rLHbeta and anti-rFSHbeta. A comparison of binding sites of anti-rLHbeta was performed using the preembeeding and the postembeeding methods. Antigenicity was observed on secretory granules whatever the method used. However, binding sites of anti-rLHbeta were detected inside the cisternae of the rough endoplasmic reticulum only with the preembedding method.

Ultrastructure of the Parotid Gland of the Nine-Banded Armadillo

1977 ◽  
Vol 35 ◽  
pp. 640-641
Author(s):  
J. R. Ruby
Keyword(s):  
Endoplasmic Reticulum ◽  
Parotid Gland ◽  
Periodic Acid ◽  
Acinar Cells ◽  
Duct System ◽  
Parotid Glands ◽  
Dasypus Novemcinctus ◽  
Anterior Portion ◽  
Periodic Acid Schiff ◽  
Thin Sections

Parotid glands were obtained from five adult (four male and one female) armadillos (Dasypus novemcinctus) which were perfusion-fixed. The glands were located in a position similar to that of most mammals. They extended interiorly to the anterior portion of the submandibular gland.In the light microscope, it was noted that the acini were relatively small and stained strongly positive with the periodic acid-Schiff (PAS) and alcian blue techniques, confirming the earlier results of Shackleford (1). Based on these qualities and other structural criteria, these cells have been classified as seromucous (2). The duct system was well developed. There were numerous intercalated ducts and intralobular striated ducts. The striated duct cells contained large amounts of PAS-positive substance.Thin sections revealed that the acinar cells were pyramidal in shape and contained a basally placed, slightly flattened nucleus (Fig. 1). The rough endoplasmic reticulum was also at the base of the cell.

Prefertilization ovule development in Capsella: the dyad, tetrad, developing megaspore, and two-nucleate gametophyte

1986 ◽  
Vol 64 (4) ◽  
pp. 875-884 ◽  
Author(s):  
Patricia Schulz ◽  
William A. Jensen
Keyword(s):  
Cell Wall ◽  
Electron Microscope ◽  
Acid Phosphatase ◽  
De Novo ◽  
Periodic Acid ◽  
Aniline Blue ◽  
Ovule Development ◽  
Periodic Acid Schiff ◽  
Fluorescent Material ◽  
Autophagic Vacuoles

Ovules of Capsella bursa-pastoris at the dyad and tetrad stages of meiosis and at the megaspore and two-nucleate stages of the gametophyte were studied with the electron microscope. The cells of the dyad and tetrad are separated by aniline blue fluorescent cross walls and receive all types of organelles and autophagic vacuoles that were present in the meiocyte. Autophagic vacuoles enclose ribosomes and organelles and show reaction product for acid phosphatase. Autophagic vacuoles and some plastids are absorbed into the enlarging vacuoles of the growing megaspore. Other plastids appear to survive meiosis and there is no evidence for their de novo origin. Some mitochondria appear to degenerate in the enlarging megaspore but others look healthy and there is no evidence for the de novo origin of mitochondria. The nucleolus of the developing megaspore becomes very large and the cytoplasm is extremely dense with ribosomes. The cell wall is thickened by an electron-translucent, periodic acid – Schiff negative, aniline blue fluorescent material and contains plasmodesmata that link the megaspore with the nucellus. The plasmalemma of the growing megaspore produces microvilluslike extensions into this wall that disappear with the formation of the two-nucleate gametophyte. Plasmodesmata disappear from the cell wall at the four-nucleate stage.

Lectins as Cytochemical Probes of the Developing Wheat Grain. V. Demonstration of Separate Polysaccharides Containing N-Acetyl-D-Glucosamine and D-Galactose in Nuclear Epidermal Cell Walls

1984 ◽  
Vol 11 (3) ◽  
pp. 179 ◽  
Author(s):  
BA Baldo ◽  
D Barnett ◽  
JW Lee
Keyword(s):  
Epidermal Cell ◽  
Cell Walls ◽  
Wheat Germ ◽  
Periodic Acid ◽  
Fluorescein Isothiocyanate ◽  
Wheat Grain ◽  
Periodic Acid Schiff ◽  
Thin Sections ◽  
Lectin Staining ◽  
Wheat Germ Lectin

Fluorescein isothiocyanate-labelled lectin from wheat-gem, which binds N-acetyl-D-glucosamine, and Griffonia simplicifolia, Arachis hypogaea and Glycine max lectins, each of which binds D-galactose, react with nucellar epidermal cell walls in thin sections of plastic-embedded developing wheat grain. Reactivity of these cell walls with periodic acid-Schiff reagent, the absence of staining with protein stains and the failure of a number of proteases and the endoglycosidases D and H to prevent the binding suggested that the lectin-reactive wall components are neither proteins nor N-glycosidically linked glycoproteins. Morphological differences in lectin staining patterns and treatment of sections with chitinase and α-galactosidase, prior to the reaction with the lectins, indicated that two separate polysaccharides are probably involved in the binding. Chitinase removed the reactivity of the nucellar epidermal cell walls for wheat-germ lectin but the binding of D-galactose-specific lectins was unimpaired. Conversely, α-galactosidase did not affect the binding of wheat-germ lectin but reactivity with the galactose-specific lectins was abolished. From the available evidence we conclude that one polysaccharide in the nucellar epidermal cell wall reacts with wheat-germ lectin and contains N-acetyl-D-glucosamine in a chitin-like structure. The other polysaccharide reacts with D-galactose- specific lectins by virtue of terminal α-D-galactose residues. Hydrolysis and subsequent chromatographic analysis of nucellar epidermal cell walls peeled from immature grains revealed the presence of D-glucosamine, D-glucose, D-galactose, D-xylose, L-arabinose and a trace of D-mannose.

scholarly journals Histochemical and Autoradiographic Studies on the Effects of Aging on the Mucopolysaccharides of the Periosteum

1959 ◽  
Vol 6 (2) ◽  
pp. 171-178 ◽  
Author(s):  
Edgar A. Tonna ◽  
Eugene P. Cronkite
Keyword(s):  
Chondroitin Sulfate ◽  
Toluidine Blue ◽  
Bone Growth ◽  
Periodic Acid ◽  
Neutral Type ◽  
Autoradiographic Study ◽  
Colloidal Iron ◽  
Periodic Acid Schiff ◽  
Schiff Reaction ◽  
Effects Of Aging

An autoradiographic study was made using S35-sulfate for the localization, distribution, and variation in the mucopolysaccharide content of the femoral periosteum of rats from birth to old age. The mucopolysaccharides were also studied histochemically, using toluidine blue O, Rinehart and Abu'l-Haj's colloidal iron method, and the periodic acid-Schiff reaction, before and after hyaluronidase treatment. Autoradiograms revealed the uptake of S35 particularly in the vicinity of the preosseous zone and adjacent osteoblasts. This labelling was highest at the period of rapid bone growth. With increasing age, the S35 uptake became progressively less. The preosseous zone showed γ-metachromatic staining at all ages after treatment with toluidine blue. Active osteoblasts were mostly orthochromatic, however, ß-metachromasia was exhibited at a later age. Abundant amounts of intra- and extracellular mucopolysaccharides of both the acid and neutral type were demonstrated in the periosteum. S35 uptake and γ-metachromasia show the presence of sulfated mucopolysaccharides, of which chondroitin sulfate predominates in the preosseous zone. Since S35 uptake is high in active osteoblasts, the inability to demonstrate metachromasia in osteoblasts may indicate either that chondroitin sulfate is liberated as fast as it is being produced, or that it may be present within the cells in a precursor form not detectable by histochemical methods.

scholarly journals METACHROMASIA INHIBITING COMPONENTS IN AMYLOID

1958 ◽  
Vol 6 (3) ◽  
pp. 181-184 ◽  
Author(s):  
BØRGE LARSEN
Keyword(s):  
Chondroitin Sulfate ◽  
Toluidine Blue ◽  
Serum Proteins ◽  
Periodic Acid ◽  
Methyl Violet ◽  
Inhibitory Effect ◽  
Blue Staining ◽  
Periodic Acid Schiff ◽  
Toluidine Blue Staining ◽  
Tissue Sections

I): The degree of inhibitory effect of serum proteins and a periodic acid Schiff-positive amyloid fraction on metachromasia resulting from a metachromatic amyloid fraction was studied and compared to the metachromatic potency of chondroitin sulfate exposed to the same type of inhibition. The metachromatic properties of chondroitin sulfate were close to those of the metachromatic amyloid fraction. II): In comparing the metachromasia obtained with toluidine blue and methyl violet, it was noted that on incubation with the periodic acid Schiff-positive amyloid fraction, amyloid metachromosia after methyl violet staining was inhibited less than after toluidine blue staining. III): It is emphasized that a competition between dye and proteins occurs in metachromatic staining reactions. IV): Inhibition of amyloid metachromasia in histological tissue sections could not be demonstrated with the techniques used.

Changes in surface morphology of pigmented retinal cells during differentiation in clonal culture

1981 ◽  
Vol 59 (1) ◽  
pp. 61-69 ◽  
Author(s):  
B. J. Crawford ◽  
Alex Yan ◽  
M. MacDonald
Keyword(s):  
Electron Microscope ◽  
Surface Morphology ◽  
Periodic Acid ◽  
Outer Edge ◽  
Gradual Change ◽  
Apical Surface ◽  
Periodic Acid Schiff ◽  
Clonal Culture ◽  
Cellular Attachment

The changes in surface morphology during the reexpression of differentiation of chick retinal pigmented epithelial cells (RPE) in clonal culture have been studied using the scanning electron microscope (SEM) and transmission electron microscope (TEM) and compared with those described in vivo. Three-week-old colonies demonstrated a gradual change in apical surface morphology along any colony radius. At the outer edge, the cell surfaces were either smooth with a few small filamentous protrusions or showed a varying number of large blebs. Toward the centre of the colony the surfaces demonstrated a gradual increase in filamentous protrusions. The apical surfaces of the most densely pigmented cells at the centre of the colony consisted mainly of small rounded protrusions. The changes in surface morphology of cells in the centre of younger colonies during redifferentiation were similar to those found along the radius of a 3-week-old colony. The results show that older colonies have all of the morphological stages of the redifferentiation process (and possibly the biochemical ones as well) arranged along any radius.The basal surfaces of all the colonies were covered by a thin acellular membrane that stained positively with periodic acid–Schiff (PAS) and which may contain fibronectins and appears to be involved in cellular attachment.

Light and electron microscopic demonstration by the PATS reaction of peripheral nerve regeneration

1989 ◽  
Vol 47 ◽  
pp. 952-953 ◽  
Author(s):  
B. Giammara ◽  
E. Anderson ◽  
P. Yates ◽  
J. Hanker
Keyword(s):  
Electron Microscopy ◽  
Toluidine Blue ◽  
Periodic Acid ◽  
Light And Electron Microscopy ◽  
Nerve Fibers ◽  
Blue Staining ◽  
Hydroxyl Groups ◽  
Distal Stump ◽  
Thin Sections ◽  
Toluidine Blue Staining

Although periodic acid-Schiff(PAS) type reactions have been applied to nervous tissues for many years, interest has centered upon staining glycolipids, principally myelin constituents such as the class of sphingolipids. The staining of these compounds such as sphingomyelin has generally been attributed to the presence of amino and hydroxyl groups on adjacent carbon atoms of carbohydrate of the sphingosine moiety. But unsaturated lipids also give the reaction and sphingolipids stain even if carbohydrate moieties are absent. This reaction has been used for staining myelin sheaths but lipid solvents must be avoided in processing the specimens. Toluidine blue staining of semi-thin sections of epoxy- embedded nerve specimens has also been widely used to study regenerating fibers after nerve transection or avulsion. A recent study was made in our laboratories of conduits (sleeves) tailored from biodegradable polyester (VicrylR) mesh to guide the reconnection of regenerating fibers from the proximal stump of a rat sciatic nerve, across an 11 mm gap, with fibers in the distal stump of the interrupted nerve. Complete reconnection of the stumps was observed as early as one month after creating the avulsive nerve injury.Comparison of transverse sections of the repaired sciatic with sections of control nerve with the toluidine blue stain, however, showed little evidence of axonal regeneration after one month (Figs. 1,3). A variation of the PAS reaction (depositing silver) for light and electron microscopy developed in our laboratories (PATS reaction, 5) was than applied to the study of the semi-thin sections of the epoxy-embedded control and repaired sciatic nerves of the same rat one month postsurgery. Correlative light and scanning electron microscopy by SEI and BEI modes could then be performed since the PATS reaction produced very satisfactory staining of the semi-thin sections (Figs. 3-5). Myelin was not stained by the PATS reaction in these specimens since the nerves had been processed with lipid solvents for epoxy embedment. Schwann cells, however, were very prominent in control but not in the repaired nerve. The inner layers of endoneurium and all pericapillaries associated with nerve fibers were intensely stained due to their reticulin content in both control and repaired nerve (Figs. 2,4). This was not unexpected because the PATS reaction employs a silver methenamine reagent. Thus, with the PATS reaction axons could be identified in sections of repaired nerve (Fig. 4) that could not be discerned with toluidine blue staining (Fig. 3). In sections of repaired nerve stained with either toluidine blue or the PATS reaction few axons or axis cylinders were observed but more were seen with the PATS stain (Figs. 3,4). In control nerve sections stained with either procedure many were seen (Figs. 1,2).

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