• Applied Microscopy
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• v.22 no.2
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• pp.118-126
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• 1992

The pathway and time course of fucose-containing glycoprotein synthesis and intracellular translocation in osteoclasts of the mice maxillary alveolar bone were investigated by electron microscopic radioautography. Male Balb-C mice weighing 17gm were anesthetized with Nembutal and injected via the external jugular vein with 2.5 mCi of $L-[6-^{3}H]-fucose$ (specific activity 16.8 mCi/mmol) in 0.1 ml of sterile saline solution. At 5, 10, 20, 35 minutes and 8 hours after administration of the $^{3}H-fucose$, animals were killed by intracardiac perfusion of 30ml of 2% glutaraldehyde in a modified Tyroid solution, pH 7.4. The maxillae were then removed and further fixed in Karnovsky fixative for an additional 3-4 hours. After rinsing in 0.1M cacodylate buffer for 10 minutes, the maxillae were demineralized for 2 weeks at $4^{\circ}C$ in ethylene diamine tetra acetate containing 2% glutaraldehyde. The first interdental areas were mesiodistally sectioned into slices of 1mm thickness and postfixed in osmium tetroxide. Tissues were then dehydrated and embedded in Poly Bed. To prepare electron microscopic radioautography, the dipping method of Kopriwa (1973) was employed. Thin sections were coated with a crystalline monolayer of ILford $L_4$ photographic emulsion. After exposure for 4 months at $4^{\circ}C$, the sections were developed Kodak Microdol-X and Phenidon (for compact grains), fixed in 30% sodium thiosulfate, stained with uranyl acetate and lead citrate and examined in the electron microscope (JEOL 1200 EX). At 5, 10 and 20 minutes after injection, $^{3}H-fucose$ was concentrated in Golgi cisternae of the osteoblasts. By 35 minutes the labels were observed over small vesicles in the suprannclear area of osteoclasts. At 8 hours, numerous silver grains were located on the ruffled border and cell membrane of osteoclasts. These results indicate that fucose molecules are added in the Golgi apparatus and small vesicles appear to be responsible for translocation of the glycoproteins to the marginal portion of osteoblasts. The glycoproteins are distributed on the osteoclast cell surface and especially over the ruffled border.

• The Korean Journal of Malacology
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• v.26 no.2
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• pp.177-184
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• 2010

The structure of the ovary, ultrastructure of oocytes and morphological characteristics of vitellogenesis during oogenesis in female Gomphina veneriformis were investigated in clams collected from coastal waters of Samchok, Gangwon-do, Kore. In the previtellogenic oocytes, the Golgi complex was involved in the formation of a number of vacuoles. In the early vitellogenic oocytes, lipid droplets appeared among the Golgi complex, endoplasmic reticulum, and mitochondria in the cytoplasm of the oocyte were involved in the formation of lipid droplets. Coated vesicles, resulting from endocytosis appeared at the basal region of the early vitellogenic oocyte. The uptake of nutritive materials in the coated vesicles formed by receptor-mediated endocytosis appeared through the formation of coated endocytotic pits on the oolemma. In the late vitellogenic oocytes, large yolk granules were formed by a combination of small yolk granules. In the mature oocyte, a mature yolk granule in composed of three components: crystaline core, electron lucent cortex, and a limiting membrane. According to cytological and histological observations, vitellogenesis occurred by way of endogenous autosynthesis and exogenous heterosynthesis. Autosynthesis involved the conbined activities of the Golgi complex, mitochondria, rough endoplasmic reticulum, whereas heterosynthesis involved endocytotic incorporation of extraovarian precursors at the basal region of the early vitellogenic oocyte. The follicle cells which was attached to oocytes, were involved in the development of the previtellogenic and early vitellogenic oocytes as a kind of nutritive cells containing a number of glycogen particles and lipid droplets in the cytoplasm.

• The Korean Journal of Physiology and Pharmacology
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• v.8 no.2
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• pp.83-88
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• 2004

The purpose of this study was to provide a basis for studying the molecular mechanism of pharmacological action of chlorhexidine digluconate. Large unilamellar vesicles (OPGTL) were prepared with total lipids extracted from cultured Porphyromonas gingivalis outer membranes (OPG). The anthroyloxy probes were located at a graded series of depths inside a membrane, depending on its substitution position (n) in the aliphatic chain. Fluorescence polarization of n-(9-anthroyloxy)stearic acid was used to examine effects of chlorhexidine digluconate on differential rotational mobility, while changing the probes' substitution position (n) in the membrane phospholipids aliphatic chain. Magnitude of the rotational mobility of the intact six membrane components differed depending on the substitution position in the descending order of 16-(9-anthroyloxy)palmitic acid (16-AP), 12, 9, 6, 3 and 2-(9-anthroyloxy)stearic acid (12-AS, 9-AS, 6-AS, 3-AS and 2-AS). Chlorhexidine digluconate increased in a dose-dependent manner the rate of rotational mobility of hydrocarbon interior of the OPGTL prepared with total lipids extracted from cultured OPG, but decreased the mobility of membrane interface of the OPGTL. Disordering or ordering effects of chlorhexidine digluconate on membrane lipids may be responsible for some, but not all of its bacteriostatic and bactericidal actions.

• BMB Reports
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• v.43 no.5
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• pp.362-368
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• 2010

Dermcidin is a human antibiotic peptide that is secreted by the sweat glands and has no homology to other known antimicrobial peptides. As an initial step toward understanding dermcidin's mode of action at bacterial membranes, we used homonuclear and heteronuclear NMR to determine the conformation of the peptide in 50% trifluoroethanol solution. We found that dermcidin adopts a flexible amphipathic $\alpha$-helical structure with a helix-hinge-helix motif, which is a common molecular fold among antimicrobial peptides. Spin-down assays of dermcidin and several related peptides revealed that the affinity with which dermcidin binds to bacterial-mimetic membranes is primarily dependent on its amphipathic $\alpha$-helical structure and its length (>30 residues); its negative net charge and acidic pI have little effect on binding. These findings suggest that the mode of action of dermcidin is similar to that of other membrane-targeting antimicrobial peptides, though the details of its antimicrobial action remain to be determined.

• Applied Microscopy
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• v.30 no.3
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• pp.311-319
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• 2000

The gill morphology and ultrastructure of the fleshy shrimp, Penaeus chinensis were investigated by light and electron microscopy. Fleshy shrimp has dendrobranchiate gills. Gill has a longitudinal septum dividing them into afferent and efferent channel. Each gill lamella is covered by multi-layered thin cuticle of different electron density. The lamella basal cell is squamous and contains cytoplasm of electron dense. Simple epithelial layer consists of squamous epithelium contained large nucleus. The lamella pillar structures are characterized by the axial microtubules and lateral membrane interdigitations Secretory cells of AB-PAS negative are multicellular gland. In active gland each cell boundary is not apparent and the cytoplasm contains smooth endoplasmic reticula, mitochondria, membrane-bounded secretory vesicles of low electron density and granular resettes. In inactive gland each cell boundary is apparent and the cytoplasm is occupied with numerous small granules of electron dense. The well-developed rough endoplasmic reticula and Golgi apparatus are observed in the unicellular gland of alcian blue positive.

• The Korean Journal of Physiology
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• v.28 no.2
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• pp.203-214
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• 1994

Using the methods of stopped-flow and epifluorescence microscopy with entrapped fluorophore, membrane permeability of $NH_3$ and weak acids in liposomes, renal brush border (BBMV) and basolateral membrane vesicles (BLMV), and primary culture cells from renal proximal tubule was measured. Permeability coefficient (cm/sec) of $NH_3$ was $(2.9{\times}10^{-2}$ in phosphatidylcholine liposome $25^{\circ}C)$, $5.9{\times}10^{-2}$ in renal proximal tubule cell $(37^{\circ}C)$, $4.0{\times}10^{-2}\;and\;2.4{\times}10^{-2}$ in BBMV and BLMV $(25^{\circ}C)$, respectively. Formic acid has the highest permeability coefficient among the weak acids tested, which was $4.9{\times}10^{-3}$ in liposome, $5.0{\times}10^{-3}$ in renal proximal tubule cell, $9.1{\times}10^{-3}$ in BBMV and $3.8{\times}10^{-3}$ in BLMV. There was a linear relationship between external concentration of nonionized formic acid and initial rate of flux of formic acid in liposome, and the slope coincided with the value of permeability coefficient of formic acid measured in pH 7.0. These results show that techniques of stopped-flow and epifluorescence microscopy with entrapped fluorophore provide the precise method of measurement of very rapid transport of nonelectrolytes through membranes with the advantages of instantaneous mixing effect, good resolution time and easy manipulation.

• Korean Journal of Veterinary Research
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• v.31 no.1
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• pp.89-97
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• 1991

For the ultrastructural observation on Babesia gibsoni(B gibsoni), the protozoa were challenged experimentally to splectomized dog. To examine the ultrastructure of the B gibsoni in the erythrocyte, the infected erythrocytes were collected at the cephalic or jugular vein of the dog. The results obtained by TEM(transmission electron microscopy) were as follows; 1. The sizes of protozoa in erythrocytes are $0.92{\pm}0.36{\mu}m{\times}0.67{\pm}0.21{\mu}m$, the sizes of nucleus of the protozoa are $0.55{\pm}0.24{\mu}m{\times}0.38{\pm}0.26{\mu}m$, and sizes of rhoptries in plasma of the protozoa are $0.33{\pm}0.05{\mu}m{\times}0.25{\pm}0.07{\mu}m$, respectively. 2. The tropozoite membrane in the erythrocyte was one, and it's nuclear membrane was made up of double. But the protozoa of initial stage in infected erythrocyte had double clear mambranes, and distinguished from plasma membrane of red blood cell. 3. The mitochondrialike structures covered with two membranes were observed in the protozoa. 4. Mitochondria and vesicles of the reticulocyte were observed near protozoa in the erythrocyte. 5. There are rhoptry, coiled structure and single nucleous in the merozoite. 6. The shape of rhoptry was round or ovoid form and in occasionally, the content of rhoptry was lost partially. 7. There was able to observe the dividing process of the protozoa. 8. Maurer's cleft-like structure was observed.

• Applied Microscopy
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• v.37 no.2
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• pp.103-109
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• 2007

The ultrastructure of submandibular gland was examined in the Korean spider shrew, Sorex caecutiens. The submandibular gland wat composed of acini and salivary ducts. A submadibular acinus was a mixed gland having serous demilune cells and mucous cells that were filled with well developed rER, mitochondria and large amount of dense secretory granules. Serous acinar granules were oval shape without distinct limiting membrane on the border and it had only coarse specks with various density. Mucous acinar granules were oval shape without distinct limiting membrane and had a variety pattern with several thin or transparent bands into the homogeneous dense matrix. Thus submandibular acinar granules of S. caecutiens belonging to subfamily Soricinae were distinct from the other mammalian species including Crocidurinae, because of the absence of limiting membrane of acinar granules and specific pattern of mucous acinar granules. Granular duct cells had large amount of small granular vesicles and several characteristic structures of granule which were revered with stratified limiting membranes and filled with coarse serous-like granule or homogeneous matrix.

• The Korean Journal of Physiology and Pharmacology
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• v.4 no.1
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• pp.63-72
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• 2000

Chronic exposure to cadmium (Cd) results in an inhibition of protein endocytosis in the renal proximal tubule, leading to proteinuria. In order to gain insight into the mechanism by which Cd impairs the protein endocytosis, we investigated the effect of Cd on the acidification of renal cortical endocytotic vesicles (endosomes). The endosomal acidification was assessed by measuring the pH gradient-dependent fluorescence change, using acridine orange or FITC-dextran as a probe. In renal endosomes isolated from Cd-intoxicated rats, the $V_{max}$ of ATP-driven fluorescence quenching ($H^+-ATPase$ dependent intravesicular acidification) was significantly attenuated with no substantial changes in the apparent $K_m,$ indicating that the capacity of acidification was reduced. When endosomes from normal animals were directly exposed to free Cd in vitro, the $V_{max}$ was slightly reduced, whereas the $K_m$ was markedly increased, implying that the biochemical property of the $H^+-ATPase$ was altered by Cd. In endosomes exposed to free Cd in vitro, the rate of dissipation of the transmembrane pH gradient after $H^+-ATPase$ inhibition appeared to be significantly faster compared to that in normal endosomes, indicating that the $H^+-conductance$ of the membrane was increased by Cd. These results suggest that in long-term Cd-exposed animals, free Cd ions liberated in the proximal tubular cytoplasm by lysosomal degradation of cadmium-metallothionein complex (CdMT) may impair endosomal acidification 1) by reducing the $H^+-ATPase$ density in the endosomal membrane, 2) by suppressing the intrinsic $H^+-ATPase$ activity, and 3) possibly by increasing the membrane conductance to $H^+$ ion. Such effects of Cd could be responsible for the alterations of proximal tubular endocytotic activities, protein reabsorption and various transporter distributions observed in Cd-exposed cells and animals.

• IMMUNE NETWORK
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• v.23 no.1
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• pp.3.1-3.14
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• 2023

Microvilli are outer membrane organelles that contain cross-linked filamentous actin. Unlike well-characterized epithelial microvilli, T-cell microvilli are dynamic similar to those of filopodia, which grow and shrink intermittently via the alternate actin-assembly and -disassembly. T-cell microvilli are specialized for sensing Ags on the surface of Ag-presenting cells (APCs). Thus, these finger-shaped microprotrusions contain many signaling-related proteins and can serve as a signaling platforms that induce intracellular signals. However, they are not limited to sensing external information but can provide sites for parts of the cell-body to tear away from the cell. Cells are known to produce many types of extracellular vesicles (EVs), such as exosomes, microvesicles, and membrane particles. T cells also produce EVs, but little is known about under what conditions T cells generate EVs and which types of EVs are released. We discovered that T cells produce few exosomes but release large amounsts of microvilli-derived particles during physical interaction with APCs. Although much is unanswered as to why T cells use the same organelles to sense Ags or to produce EVs, these events can significantly affect T cell fate, including clonal expansion and death. Since TCRs are localized at microvilli tips, this membrane event also raises a new question regarding long-standing paradigm in T cell biology; i.e., surface TCR downmodulation following T cell activation. Since T-cell microvilli particles carry T-cell message to their cognate partner, these particles are termed T-cell immunological synaptosomes (TISs). We discuss the potential physiological role of TISs and their application to immunotherapies.