• Title/Summary/Keyword: filopodia

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Localization of cytoskeletal proteins in Pneumocystis carinii by immuno-electron microscopy

  • Yu, Jae-Ran;Pyon, Jae-Kyong;Seo, Min;Jung, Byung-Suk;Cho, Sang-Rock;Lee, Soon-Hyung;Hong, Sung-Tae
    • Parasites, Hosts and Diseases
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    • v.39 no.1
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    • pp.13-21
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    • 2001
  • Pneumocystis carinii causes serious pulmonary infection in immuno-suppressed patients. This study was undertaken to observe the cytoskeletal proteins of P. carinii by immune-electron microscopy. P. carinii infection was experimentally induced by immunosuppression of Sprague-Dawley rats for seven weeks, and their lungs were used for the observations of this study. The gold particles localized actin, tropomyosin, and tubulin. The actin was irregularly scattered in the cytoplasm of the trophic forms but was much more concentrated in the inner space of the cell wall of the cystic forms called the inner electron-lucent layer No significant amount of tropomyosin was observed in either trophic forms or cystic forms. The tubulin was distributed along the peripheral cytoplasm and filopodia of both the trophic and cystic forms rather than in the inner side of the cytoplasm. Particularly, in the cystic forms, the amount of tubulin was increased and located mainly in the inner electron-lucent layer of the cell wall where the actin was concentrated as well. The results of this study showed that the cell wall of P carinii cystic forms is a structure whose inner side is rich in actin and tubulin. The location of the actin and tubulin in P. carinii suggests that the main role of these proteins is an involvement in the protection of cystic forms from the outside environment by maintaining rigidity of the cystic forms.

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Regulation of retinal angiogenesis by endothelial nitric oxide synthase signaling pathway

  • Ha, Jung Min;Jin, Seo Yeon;Lee, Hye Sun;Shin, Hwa Kyoung;Lee, Dong Hyung;Song, Sang Heon;Kim, Chi Dae;Bae, Sun Sik
    • The Korean Journal of Physiology and Pharmacology
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    • v.20 no.5
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    • pp.533-538
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    • 2016
  • Angiogenesis plays an essential role in embryo development, tissue repair, inflammatory diseases, and tumor growth. In the present study, we showed that endothelial nitric oxide synthase (eNOS) regulates retinal angiogenesis. Mice that lack eNOS showed growth retardation, and retinal vessel development was significantly delayed. In addition, the number of tip cells and filopodia length were significantly reduced in mice lacking eNOS. Retinal endothelial cell proliferation was significantly blocked in mice lacking eNOS, and EMG-2-induced endothelial cell sprouting was significantly reduced in aortic vessels isolated from eNOS-deficient mice. Finally, pericyte recruitment to endothelial cells and vascular smooth muscle cell coverage to blood vessels were attenuated in mice lacking eNOS. Taken together, we suggest that the endothelial cell function and blood vessel maturation are regulated by eNOS during retinal angiogenesis.

Astaxanthin induces migration in human skin keratinocytes via Rac1 activation and RhoA inhibition

  • Ritto, Dakanda;Tanasawet, Supita;Singkhorn, Sawana;Klaypradit, Wanwimol;Hutamekalin, Pilaiwanwadee;Tipmanee, Varomyalin;Sukketsiri, Wanida
    • Nutrition Research and Practice
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    • v.11 no.4
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    • pp.275-280
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    • 2017
  • BACKGROUND/OBJECTIVES: Re-epithelialization has an important role in skin wound healing. Astaxanthin (ASX), a carotenoid found in crustaceans including shrimp, crab, and salmon, has been widely used for skin protection. Therefore, we investigated the effects of ASX on proliferation and migration of human skin keratinocyte cells and explored the mechanism associated with that migration. MATERIAL/METHOD: HaCaT keratinocyte cells were exposed to $0.25-1{\mu}g/mL$ of ASX. Proliferation of keratinocytes was analyzed by using MTT assays and flow cytometry. Keratinocyte migration was determined by using a scratch wound-healing assay. A mechanism for regulation of migration was explored via immunocytochemistry and western blot analysis. RESULTS: Our results suggest that ASX produces no significant toxicity in human keratinocyte cells. Cell-cycle analysis on ASX-treated keratinocytes demonstrated a significant increase in keratinocyte cell proliferation at the S phase. In addition, ASX increased keratinocyte motility across the wound space in a time-dependent manner. The mechanism by which ASX increased keratinocyte migration was associated with induction of filopodia and formation of lamellipodia, as well as with increased Cdc42 and Rac1 activation and decreased RhoA activation. CONCLUSIONS: ASX stimulates the migration of keratinocytes through Cdc42, Rac1 activation and RhoA inhibition. ASX has a positive role in the re-epithelialization of wounds. Our results may encourage further in vivo and clinical study into the development of ASX as a potential agent for wound repair.

Rac1 inhibition protects the kidney against kidney ischemia/reperfusion through the inhibition of macrophage migration

  • You Ri Park;Min Jung Kong;Mi Ra Noh;Kwon Moo Park
    • The Korean Journal of Physiology and Pharmacology
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    • v.27 no.3
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    • pp.257-265
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    • 2023
  • Kidney ischemia/reperfusion (I/R) injury, a common cause of acute kidney injury (AKI), is associated with the migration of inflammatory cells into the kidney. Ras-related C3 botulinum toxin substrate 1 (Rac1), a member of the Rho family of small GTPase, plays an important role in inflammatory cell migration by cytoskeleton rearrangement. Here, we investigated the role of Rac1 on kidney I/R injury and macrophage migration. Male mice were subjected to either 25 min of bilateral ischemia followed by reperfusion (I/R) or a sham operation. Some mice were administrated with either NSC23766, an inhibitor of Rac1, or 0.9% NaCl (vehicle). Kidney damage and Rac1 activity and expression were measured. The migration and lamellipodia formation of RAW264.7 cells, mouse monocyte/macrophage, induced by monocyte chemoattractant protein-1 (MCP-1, a chemokine) were determined using transwell migration assay and phalloidin staining, respectively. In sham-operated kidneys, Rac1 was expressed in tubular cells and interstitial cells. In I/R-injured kidneys, Rac1 expression was decreased in tubule cells in correlation with the damage of tubular cells, whereas Rac1 expression increased in the interstitium in correlation with an increased population of F4/80 cells, monocytes/macrophages. I/R increased Rac1 activity without changing total Rac1 expression in the whole kidney lysates. NSC23766 administration blocked Rac1 activation and protected the kidney against I/R-induced kidney damage and interstitial F4/80 cell increase. NSC23766 suppressed monocyte MCP-1-induced lamellipodia and filopodia formation and migration of RAW 264.7 cells. These results indicate Rac1 inhibition protects the kidney against I/R via inhibition of monocytes/macrophages migration into the kidney.

T Cell Microvilli: Finger-Shaped External Structures Linked to the Fate of T Cells

  • Hye-Ran Kim;Jeong-Su Park;Won-Chang Soh;Na-Young Kim;Hyun-Yoong Moon;Ji-Su Lee;Chang-Duk Jun
    • 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.

The Morphological Study on Inflammation of Murine Knee Joint by Lipopolysaccharide - Based on the Morphological Changes of Synovial Membrane and Fibrous Membrane - (Lipopolysaccharide로 유발된 생쥐 무릎관절낭 염증에 관한 형태학적 연구 - 윤활관절막과 섬유관절막의 변화를 중심으로 -)

  • Kim, Jin-Tack;Ahn, Sang-Hyun;Choi, Nan-Hee;Chung, Jae-Man;Park, In-Sick;Gang, Yun-Ho;Kim, Ho-Hyun;Lee, Hai-Poong
    • The Journal of Dong Guk Oriental Medicine
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    • v.7 no.2
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    • pp.107-120
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    • 1999
  • Synovial joint of BALB/C mice were injeced with Lipopolysaccharide(LPS) were observed to investigate the morphological changes of synovial capsule caused by rheumatoid arthritis(RA). The RA on female Balb/c mice were induced by LPS injection, as dose of $300{\mu}{\ell}/kg$, into synovial cavity of knee joint. And then these specimen were fixed in 10% neutral buffered formalin and were decalcificated in EDTA solution for 4 weeks. The hyperplasia of synovium were appeared in synovial membrane. The filopodia of phagocytic like synoviocyte(type I synoviocyte) projected into synovial cavity and the number of fibroblast like synoviocyte(type II synoviocyte) with well-developed endoplasmic reticulum were increased in synovium. In fibrous membrane, the fibrosis induced by synthesis of collagen fiber were enlarged to all fibrous membrane, and the number of fibroblast were increased. A great number of inflammation component cell as lymphocyte and neutrophil leukocyte were infiltrated around capillary and the degranulate typed mast cell were increased. As results indicated that the hyperplasia of synovium induced by LPS, subsequently to cause the fibrosis, infiltration of imflammation component cell, and increase of degranulated type mast cell as same as symptoms of RA.

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Studies on the Visceral Ganglion and Right Parietal Ganglion in the African Giant Snail, Achatina fulica II. Ultrastructural Method (아프리카 왕달팽이 (Achatina fulica) 내장신경절 및 우 체벽신경절에 관한 연구 II. 미세구조적 방법)

  • Chang, Nam-Sub
    • Applied Microscopy
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    • v.31 no.1
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    • pp.101-108
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    • 2001
  • Five kinds of neurosecretory cells (type-A, B, C, D and E) and neuropiles surrounding them were observed in the visceral ganglion and the right parietal ganglion of the African giant snail, Achatina fulica, by transmission electron microscopy. Type-A cells (diameter, $35{\mu}m$) are the most popular cells in the cortex of the two ganglions, which are of triangular or irregular forms. In their cytoplasm, there are found large granules of 1 fm in diameters and small round granules of about $0.1{\mu}m$ in diameters. Small granules are classified into the ones of high electron density and the others of middle electron density. Type-B cells (diameter, $19\times12{\mu}m$) are evenly distributed over various portions of cortex and medulla of the two ganglions. They are similar to type-A cells in shapes. The cytoplasm of type-B cells is crowded with high electron dense granules of about $0.1{\mu}m$. Round granules of about $0.7{\mu}m$ in diameters are also found but rarely. Type-C cells are the smallest cells whose sizes are about $8\times6{\mu}m$. Each of them contains a large nucleus of about $6\times5{\mu}m$. Its cytoplasm is full of electron dense granules of about $0.23{\mu}m$, each of which is artually an assembly of tiny granules of about $0.03{\mu}m$. Type-D cells are middle-size cells of about $28\times20{\mu}m$, which take ellipsoidal or irregular forms. They are found in the cortex more than in the medulla. Their cytoplasm looks dark due to the high electron density and, in it, two kinds of round granules whose sizes are $1.6{\mu}m$fm and $0.6{\mu}m$, respectively, are observed. Type-E cells are large cells of about $100\times50{\mu}m$, which are rarely found in the upper and middle portions of the two ganglions. The nucleus of the cell, which is very large $(70\times30{\mu}m)$ for the cytoplasm, contains electron dense round granules of diverse sizes (diameters, $1\sim0.2{\mu}m$). The surface of the cell protrudes filopodia of various forms and phagocytizes decrepit cells. Neuropiles are surrounding the neurosecretory cells. In nerve fibers, synaptic vesicles are observed, which are classified into six classes according to their electron densities , sizes and shapes.

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An Experimental Studies on the alleviation effects of Daebangpoongtang(大防風湯) in LPS-induced arthritis (대방풍탕(大防風湯)의 LPS유발(誘發) 관절염(關節炎) 완화효과(緩和效果)에 관한 실험적(實驗的) 연구(硏究))

  • Jeon, Won-June;Shin, Gil-Jo;Lee, Won-Chul
    • The Journal of Dong Guk Oriental Medicine
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    • v.9
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    • pp.35-49
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    • 2000
  • Purpose: This study is to investigate the alleviation effects of Daebangpoongtang in LPS induced arthritis in mice knee joint. Methods : Daebangpoongtang was chosen to treat the arthritis caused by injecting $300{\mu}g/kg$ LPS to mice knee joint. The control group had no treatment, while the LPS group was injected $300{\mu}g/kg$ LPS to mice knee joint and the DBP group was oral administrated of Daebangpoongtang. After injection of $300{\mu}g/kg$ LPS to mice knee joint, the alteration of synovial lining cell, vessel, fibrosis, distribution of collagen fiber, fibroblast, mast cell, infiltration of inflammation component cell and distribution of ICAM and VCAM was observed by light microscope(BX50). Results : In the DBP extract treatment group, the distribution of vessel, the enlargement of synovial lining cell layer, the synovial lining cells with filopodia, the fibrosis, the distribution of fibroblast in synovial membrane, the distribution of TCAM and VCAM on the knee joint was less than that of LPS group. Infiltrated lymphocyte into the apical surface had not observed in the DBP extract treatment group. The distribution of mast cell was as same as control group(no treatment group) and it showed granulated type. Conclusion: According to the above results, it might be considered that the administration of Daebangpoongtang has a curative effect on synovial membrane injury in arthritis by inhibiting increase of vessel, cell adhesion molecule(ICAM and VCAM) in LPS induced arthritis.

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Comparative Studies on the Ultrastructure of Salivary Ducts between the Two Species of Snails, Achatina fulica and Incilaria fruhstorferi (두 종의 달팽이류 (Achatina fulica and Incilaria fruhstorferi) 사이의 타액관의 미세구조에 관한 비교연구)

  • Chang, Nam-Sub;Han, Jong-Min;Kim, Sang-Won;Lee, Kwang-Ju;Hwang, Sun-Jong
    • Applied Microscopy
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    • v.30 no.1
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    • pp.89-100
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    • 2000
  • We observed the salivary ducts of two species of snails, Achatina fulica and Incilaria fruhstoferi with an electron microscope, and obtained the following results. The intralobular and interlobular ducts of Achatina fulica assume the forms of round or ellipsoidal doughnuts. The boundaries between the endothelial cells are not clear. It is also found that the cytoplasm of the endothelial cells consists of the membrane infolded in interdigital form, and there are well -developed microvilli at the apical portion of the cytoplasm. On the other hand, the intralobular and interlobular ducts of Incilaria fruhtoferi consist of the irregular simple columnar epithelia. The high electron dense cytoplasm is filled with the irregular round granules. The microvilli at the apical portion of the cytoplasm are not so well-developed as those in Achatina fulica. In the salivary duct of Achatina fulica, the lumen has narrow and long tubular structure. The boundaries between the endothelial cells are not clear. The cytoplasm is full of many vacuoles and electron lucent granules. At the apical portion of the cytoplasm, lots of short and thin microvilli are found. The salivary duct of Incilaria fruhstorferi is wider ($65\times250{\mu}m$ in diameter) than that of Achatina fulica, and consists of endothelial cells of the same structures. At the apical portion of those endothelial cells, a lot of junction apparatus such as desmosomes are observed. The vessels in the salivary ducts of Achatina fulica and Incilaria fruhstoferi are observed mainly in the connective tissues between the salivary glands. The endothelial cell of the vessel has the irregular structure and looks dark due to the high electron density. These cells protrude their filopodia and phagocytosize foreign bodies.

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The Morphological Study on the Effects of Ginseng under Stress - Histochemical and Electron Microscopic Observations of the Rat Adrenal Cortex - (스트레스에 대한 인삼효과의 형태학적 연구 - 흰쥐 부신피질의 조직화학적 및 전자현미 경석 관찰 -)

  • Lee, Jae-Hyun
    • Current Research on Agriculture and Life Sciences
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    • v.3
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    • pp.190-196
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    • 1985
  • In order to know the effects of ginseng on the stress, histochemical and electron microsbopic observations were carried out of the adrenal cortex on the stressed rats. The experiments were carried out as follow : non-stressed after saline injection (S-NS-AD), stressed after saline injection (S-S-AD), non-stressed after ginseng injection (G-NS-AD), stressed after ginseng injection (G-S-AD). Saline and ginseng extract were treated by peritoneal injection for 30 days. The results obtained are summarized as follows : Histologically a number of small lipid droplets are found more in S-S-AD group than in S-NS-AD group, whereas in ginseng treated groups numerous large lipid droplets are found. Histochemically cholesterol and its ester, glycolipid, phospholipid and glycogen are highly found in ginseng-treated groups. On the other hand, glycogen and cholesterol are also observed in S-S-AD group. Electron microscopically a numbers of lipid droplets are observed in S-S-AD, G-NS-AD and G-S-AD group, especially large lipid droplets and moderate glycogen are found in G-NS-AD and G-S-AD group. In ginseng-treated groups and S-S-AD group, dilated SER, extended intercellular space and dense bodies are observed, and the filopodiae are found in G-S-AD group. These data suggest that ginseng was activate the metabolism and synthesis of the adrenal cortical cells on the stress.

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