A New Electron Microscopic Histo-Cytochemical Staining Method -Demonstration of Glycogen Particles-

Shinji, Yuji; Shinji, Eiko; Mizuhira, Vinci

doi:10.1267/ahc.8.139

Cited by 23 publications

(16 citation statements)

References 9 publications

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“…Sections were stained with uranyI acetate and lead citrate, or with bismuth nitrate according to the method of Shinji et al (1975) and viewed with a JEOL lOOSX transmission electron microscope at an accelerating voltage of SO kV.…”

Section: Methodsmentioning

confidence: 99%

Ultrastructure of proteinase-secreting cells of Candida albicans studied by alkaline bismuth staining and immunocytochemistry

Akashi

Homma

Kanbe

et al. 1993

Journal of General Microbiology

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The ultrastructure of Cundida ulbicans cells induced to secrete extracellular proteinase (EPR) has been studied. Electron microscopy employing alkaline bismuth staining, a method which stains polysaccharides, clearly revealed Golgi-like bodies and secretory vesicles in C. albicans cells. After EPR induction, there was no apparent increase in the number of these structures. Instead, many flocculent granules appeared at the periphery of induced cells. The granules were similar to secretory vesicles in size, but were more irregular in shape. Similar granules were observed in non-induced cells, though less frequently than in induced cells. Brefeldin A, a specific inhibitor of membrane transport in the secretory pathway, caused the accumulation of EPR and Golgi-like bodies in EPR-induced cells, but did not affect the accumulation of the granules, These results suggest that the granules are unrelated to EPR secretion. Electron microscope immunocytochemistry with affinity-purified anti-EPR antibodies showed that the granules in EPR-induced cells were recognized by the antibodies. This recognition was completely inhibited by the presence of glycogen, suggesting that antibodies cross-react with glycogen-like polysaccharides in the granules. Although the location of EPR within the cells remains unclear, the results suggest that EPR might be secreted via the constitutive secretory pathway, and that EYR is glycosylated to give a structure with some similarity to glycogen.

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Section: Methodsmentioning

confidence: 99%

Ultrastructure of proteinase-secreting cells of Candida albicans studied by alkaline bismuth staining and immunocytochemistry

Akashi

Homma

Kanbe

et al. 1993

Journal of General Microbiology

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“…Some ultra-thin sections were treated for alkaline bismuth staining for glycogen granules (Shinji et al 1975, Shinji et al 1976) with a minor modification as follows. Ultrathin sections placed on copper grids were exposed for 40 min at 40 ° C in alkaline bismuth solution, which consists of 0.5% sodium hydroxide, 0.2% potassium sodium tartarate and 0.1% bismuth subnitrate (Nacalai, Kyoto, Japan).…”

Section: Transmission Electron Microscopymentioning

confidence: 99%

Organogenesis of the juxta‐oral organ in mice

Ito

Nakashima²,

Yoshioka

et al. 2009

Journal of Anatomy

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The juxta-oral organ is a bilateral organ in the mammalian bucca. It consists of epithelial cords with surrounding mesenchyme. It develops from embryonic oral epithelium, but its macroscopic morphology in mice is less studied and seems to be very different from that of humans. The juxta-oral organ in mice extends more widely from the subcutaneous tissue of the mandible near the lateral fascia of the masseter to the submucosa of the soft palate. In this paper, we report that the mutant mouse allele Bmp7 lacZ presented intense lacZ expression in the epithelial component of the juxta-oral organ in its homo-and heterozygous states. The main aims of this study were to show that this mutant mouse allele is suitable for observing macroscopic structure of the juxta-oral organ and to describe the development of this organ during embryonic and postnatal stages. Whole-mount β-gal staining of this strain of mouse showed that the juxta-oral organ in mice appeared at E12.0 from oral epithelium and lost connection with it before E12.5. Then, the juxta-oral organ extended anteriorly to the lateral fascia of the masseter and posteriorly to the submucosal layer of the soft palate via the orbit. The mature juxta-oral organ had no connection to other epithelia such as those of the bucca and parotid duct. It persisted until adulthood and there seemed to be no tendency to regress. Transmission electron microscopy showed that each part of the juxta-oral organ was an epithelial cord surrounded by a basement membrane and mesenchymal tissue.

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“…Thin sections were cut with a diamond knife on an ultramicrotome (OmU4; Reichert). Sections were stained with uranyl acetate and lead citrate, or, for polysaccharide staining, with bismuth nitrate by the method of Shinji et al (1975) and viewed with a transmission electron microscope (100SX; JEOL, Tokyo, Japan) at an accelarating voltage of 80 kV. For immunocytochemistry, sections on nickel grids were treated with 1% (v/v) H202 for 30 min.…”

Section: Electron Microscopymentioning

confidence: 99%

Localized accumulation of cell wall mannoproteins and endomembranes induced by brefeldin A in hyphal cells of the dimorphic yeastCandida albicans

1997

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Summary.Candido albicans, a dimorphic yeast, has the abililty to switch its growth form between budding growth and hyphal growth. Since fungal growth involves secretory processes, spatial control of secretion should play a crucial role in such a morphogenetic transition. Brefeldin A (BFA), an inhibitor of the membrane trafficking system of eukaryotes, increases the occurrence of Golgi-like cisternae in the yeast. In the present study, BFA was used to obtain further insights into the spatial organization of secretory processes in hyphal growth of C. albicans. BFA completely inhibited the formation and growth of germ tubes at a concentration of 35 .aM or higher. Electron microscopy of BFA-untreated germinated cells revealed many vesicles in the apical region and Golgi-like eistemae in the cytoplasm. In cells treated with 35 .aM BFA, the vesicles disappeared from the apical region, and, instead, stacked membrane cisternae and membrane-enclosed spherical dense bodies accumulated in the subapical region. These accumulated structures were positive for both polysaccharide staining and immunocytochemical staining with antibodies raised against cell surface antigens of C. albicans, as were Golgi cisternae in BFA-untreated cells. In cells treated with a higher concentration of BFA (140 `aM), the structures that appeared in cells treated with 35 aM BFA were no longer observed and the endoplasmic reticulum was extended and positive for polysaccharide staining. These results suggested that BFA affects different steps of membrane trafficking in a concentration-dependent manner. The accumulated structures induced by 35 ,aM BFA seemed to be the altered forms of Golgi cistemae. Their accumulation in the subapical region of the germ tube might indicate that tile step(s) in membrane trafticking that are associated with the Golgi pathway are vectorially organized in hyphal growth of C. albicans.

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A New Electron Microscopic Histo-Cytochemical Staining Method -Demonstration of Glycogen Particles-

Cited by 23 publications

References 9 publications

Ultrastructure of proteinase-secreting cells of Candida albicans studied by alkaline bismuth staining and immunocytochemistry

Ultrastructure of proteinase-secreting cells of Candida albicans studied by alkaline bismuth staining and immunocytochemistry

Organogenesis of the juxta‐oral organ in mice

Localized accumulation of cell wall mannoproteins and endomembranes induced by brefeldin A in hyphal cells of the dimorphic yeastCandida albicans

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