• Title/Summary/Keyword: microtubules

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Tegumental Ultrastructures of Spirometra erinacei by Developmental stages (만손열두조충의 발육단계별 표피 미세구조)

  • Sohn, Woon-Mok;Lee, Jin-Ha
    • Applied Microscopy
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    • v.35 no.1
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    • pp.41-56
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    • 2005
  • Present study was performed to observe the tegumental ultrastructures by the developmental stages which derived from the experimental life cycle of Spirometra erinacei in laboratory conditions. In SEM view, coracidium was spherical in shape with numerous cilia, and its surface was covered with long cilia, tuberclelike projections with millet-like processes, and small holes. The body surface of procercoid was covered with numerous pointed microtriches except that of frontal pit with stout spine-like ones. However that of cercomer was covered with somewhat sparse blunt-tiped microtriches. Plerocercoids of 3 days old resembled the mature procercoid in shape, and their frontal pits were covered with numerous stout spine-like microtriches. However frontal pit and body surface in more than 5 days old ones were covered with conoid microtriches. On the surface of adult scolex, hairly long filamentous and stout short microtriches were mixedly distributed. Filamentous microtriches were more densely distributed in the anterior portion than in the posterior of scolex. The neck and immature proglottid were covered with only stout short conoid microtriches. In TEM view of coracidia, embryophore and oncosphere were obviously distinguished. The embryophore contained numerous glycogen particles, mitochondria and lipid granules. The cilia on the surface of embryophore rooted in the coracidial sheath, and consisted of 9 pairs of microtubules and 2 core complex. The oncosphere was covered with a thin and unarmed tegument, and was multi-nucleated. The protoplasmic layer of procercoid and plerocercoid consisted of disc-shaped bodies, vacuoles and mitochondria. Their tegumental cells commonly retained a nucleus, granular endoplasmic reticulums and secretory granules. The protoplasmic layer of plerocercoid was more compacted than that of procercoid. From the above results, it was confirmed that the tegumental ultrastructures are something different according to the developmental stages of S. erinacei.

THE ANTICANCER EFFECT OF PACLITAXEL($Taxol^{(R)}$) IN ORAL SQUAMOUS CELL CARCINOMA XENOGRAFT (이종 이식된 구강편평세포 암종에서 Paclitaxel ($Taxol^{(R)}$)의 항암 효과)

  • Kim, Ki-Hwan;Kim, Chul-Hwan;Han, Se-Jin;Lee, Jae-Hoon
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.28 no.2
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    • pp.95-110
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    • 2006
  • The treatment for oral and maxillofacial carcinoma with chemotherapeutic agents is evaluated by many effective methods to reduce the tumor mass and cancer cell proliferation. However these chemotherapy have many serious side effects, such as bone marrow suppression, renal toxicity, G-I troubles. Therefore a possible approach to develop a clinically applicable chemotherapeutic agent is to screen anticancer activity of Taxol which is known to have very little side effect and have been used to breast cancer and ovarian carcinoma. Taxol is a new anti-microtubular anti-cancer agent extracted from the bark of the Pacific yew, Taxus brevifolia. Paclitaxel(Taxol) acts by promoting tubulin polymerization and over stabilizing microtubules agianst depolymerization. Despite the constant improvements of methods of the cancer treatment especially chemotherapy, the rate of cancer metastasis and recurrent are not decreased. Thus the investigation of new drug which have very little side effect and a possible clinically application continues to be a high priority. Considering that the Taxol have shown very effective chemotherapeutic agent with relatively low toxicity in many solid tumors, it deserves to evaluate its efficacy in oral squamous cell carcinoma. In this study, to investigate the in-vivo and in-vitro anti-cancer efficacy of Taxol in oral squamous cell carcinoma and lastly, the potency of Paclitaxel in the clinical application for oral cancer was evaluated. In vivo study, after HN22 cell line were xenografted in nude mice, the growth of tumor mass was observed, 3 mg/Kg taxol was injected intraperitoneally into nude mice containing tumor mass. The methods of these study were measurement of total volume of tumor mass, histopathologic study, immunohistochemical study, drug resistance assay, growth curve, MTT assay, flow cytometry, cDNA microarray in vivo and in vitro. The results were obtained as following. 1. The visual inspection of the experimental group showed that the volume of the tumor mass was slightly decreased but no significant difference with control group. 2. Ki-67 index was decreased at weeks 4 in experimental group. 3. Microscopic view of the xenografted tumor mass showed well differentiated squamous cell carcinoma and after Taxol injection, some necrotic tissue was seen weeks 4. 4. The growth curve of the tumor cells were decreased after 1day Taxol treatment. 5. According to the MTT assay, HN22 cell line showed relative drug resistancy above $5\;{\mu}g/ml$ concentrations of Taxol. 6. In drug resistance assay, the decrease of cell counts was seen relatively according to concentration. 7. In Flow cytometry, G2M phase cell arrests were seen in low concentration of the Taxol, while S phase cell arrests were seen in high concentration of the Taxol. 8. Using cDNA microarray technique, variable gene expression of ANGPTL4, TXNRD1, FAS, RRAGA, CTGF, CYCLINEA, P19, DUSP5, CEBPG, BTG1 were detacted in the oral squamous cell carcinoma cell after taxol treatment. In this study paclitaxel is effective against oral squamous cell carcinoma cell lines in vitro, but week effect was observed in vivo. So we need continuous study about anticancer effect of taxol in vivo in oral squamous cell carcinoma.

Turion as Dormant Structure in Spirodela polyrhiza (개구리밥 휴면구조 잠아의 구조적 특성)

  • Kwak, Mi-Young;Kim, In-Sun
    • Applied Microscopy
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    • v.38 no.4
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    • pp.307-314
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    • 2008
  • Hydrophytes such as Spirodela polyrhiza form dormant turions to withstand cold winters. The turion is an anatomically distinct structure from which a vegetative frond arises later during germination. The turions sink to the bottom of the pond when temperatures drop and remain there throughout the winter. In the spring, they float to the surface and germinate into a new frond from the turion primordium. Unlike fronds, turions are known to possess small aerenchyma, starch grains, and relatively dense cytoplasm. These features allow the turions to survive the cold winter season at the bottom of the pond. Spirodela polyrhiza has been investigated previously to a great extent, especially in its physiological, biochemical and ecological attributes. However, a little is known about the structural features of the frond and turion during turion development. Thus, the aim of the present study was to reveal the structural characteristics of the frond and turion with regard to tissue differentiation, aerenchyma development, starch distribution, and ultrastructure, with the use of electron microscopy. A moderate degree of mesophyll tissue differentiation was found in the frond, whereas the turion did not exhibit such differentiation. Within the frond tissue, approximately $37{\sim}45%$ of the cellular volume was occupied by a large aerenchyma, but only $9{\sim}15%$ was taken up by the aerenchyma in the turion. The turion cells, especially those of the turion primordium, were derived from frond cells, and contained cytoplasm. Their cytoplasm was densely packed with plastids, mitochondria, endoplasmic reticulum, Golgi bodies, and microtubules. Plasmodesmata were also well developed within these cells. The most striking feature observed was the distribution of starch grains within the plastids of turion cells. Before the turion sank to the bottom of the pond, a considerable amount of starch accumulated in the plastid stroma. The starch grains dissolved when temperatures rose in the spring, and this promptly provided the nutrients which the primordium needed for turion germination. The turion therefore, was an appropriate dormant structure for free-floating, reduced hydrophytes like Spirodela polyhriza due to its small aerenchyma and large starch grains that aided in the purpose of sinking below the surface of the water to survive cold winters. The new fronds that arose from such turions grew rapidly in the spring, beginning the new life cycle.

JSAP1 Interacts with Kinesin Light Chain 1 through Conserved Binding Segments (JSAP1과 Kinesin Light Chain 1의 결합 및 결합부위 규명)

  • Kim, Sang-Jin;Lee, Chul-Hee;Park, Hye-Young;Yea, Sung-Su;Jang, Won-Hee;Lee, Sang-Kyeong;Park, Yeong-Hong;Cha, Ok-Soo;Moon, Il-Soo;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.17 no.7 s.87
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    • pp.889-895
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    • 2007
  • A conventional kinesin, KIF5/kinesin-I, is composed of two kinesin heavy chains (KHCs) and two kinesin light chains (KLCs) and binds directly to microtubules. KIF5 motor mediates the transport of various membranous organelles, but the mechanism how they recognize and bind to a specific cargo has not yet been completely elucidated. Here, we used the yeast two-hybrid system to identify the neuronal protein(s) that interacts with the tetratricopeptide repeats (TRP) of KLCI and found a specific interaction with JNK/stress-activated protein kinase-associated protein 1 (JSAP1/JIPP3). The yeast two-hybrid assay demonstrated that the TRP 1,2 domain-containing region of KLCI mediated binding to the leucine zipper domain of JSAP1. JSAP1 also bound to the TRP region of lac2 but not to neuronal KIF5A, KIF5C and ubiquitous KIF5B in the yeast two-hybrid assay. In addition, these proteins showed specific interactions in the GST pull-down assay and by co-immunoprecipitation. KLCI and KIF5B interacted with GST-ISAP1 fusion proteins, but not with GST alone. An antibody to JSAPI specifically co-immunoprecipitated KIF5s associated with JSAP1 from mouse brain extracts. These results suggest that JSAP1, as KLC1 receptor, is involved in the KIF5 mediated transport.

Ultrastructural Changes of the Vas Deferens Epithelium by Season in a Slug Incilaria fruhstorferi (산민달팽이(Incilaria fruhstorferi) 수정관 내강 상피조직의 계절에 따른 미세구조적 변화)

  • 장남섭;정계현;한종민
    • The Korean Journal of Zoology
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    • v.39 no.4
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    • pp.400-409
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    • 1996
  • A study on the ultrastructural changes In the epithellum of the vas deferens by season was conducted for the spdng and summer specimens of a slug Incilarfa fruhstoiferl. The vas deferens of the spdng spedmen was muscular tube about 0.4mm in diameter. Its lumen was divided into three flat grooves and the each groove was subdivided into two subbranches. The luminal epithellal celis of the Vas deferens which were irregular In shape showed strong methylenophilla in a double stain of methylene blue and basic fuchsln. The lumen of the vas deferens was filled with components strongly stained by methylene blue. The circular muscle layers surrounding the luminal epithellum of the vas deferens contained numerous granules arranged at regular intervals. The vas deferens of the summer specimen also was a thick muscular tube showing 0.4 mm in diameter. Its lumen was divided into four grooves but, the each of the grooves was not subdivided to form certain branclees unlikely to the spdng spedmen. The lining epfthelium of the lumen was consisted of simple ciliated columnar cells, irregular columnar cells and conical cells. The histological features were quiet different from those of the spring spedmen which showed irregular cell arrangement. According to electron microscopy the epithelium of the vas deferens in the spring specimen was composed of irregular columnar cells which had irregular shaped nuclei. The nuclei of the epitheilal cells were relatively large in comparison to their cytoplasm. The overall electric density of the cytoplasm was relatively high. The lumen of the vas deferens in the summer specimen was lined by a epfthelium with tail ciliated columnar cells and irregular cells. The unclei of the epithellal cells were long ellipsoid or irregular in shape. Both of the cytoplasm and the nuclei were showed low electric density. in consideration with the observable cell organelles were only ndoplasmic reticulum, lysosomes and microtutules, the cell organelles were poorly developed. The apical surfaces of the epithelial cells possessed brush borders with numerous microvilli and cilia with 9+2 arrangement of microtubules. The circular muscle layers surrounding the epithelium are usually thick and the degree of development of the circular muscle layers seems to be even in the both of the spring and summer specimens.

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APOPTOTIC EFFECT IN COMBINATION OF CYCLOSPORIN A AND TAXOL ON ORAL SQUAMOUS CELL CARCINOMA CELL LINE THROUGH THE PI-3 KINASE/AKT1 PATHWAY (구강 편평세포암종 세포주에서 Cyclosporin A와 Taxol 투여시 PI-3 kinase/Akt1 Pathway에 의한 세포사멸 병용효과)

  • Kim, Kyu-Young;Lee, Jae-Hoon
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.33 no.5
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    • pp.426-436
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    • 2007
  • Oral cancer take up 2-6% of all carcinomas and squamous cell carcinoma, which is the most common type in oral cancer, has a poor prognosis due to its high metastasis and recurrence rates. In treating oral cancer, chemotherapy to the primary, metastasized and recurrent lesion is a very important and useful treatment, even though its widespread usage is limited due to high general toxicity and local toxicity to other organs. Taxol, a microtubule stabilizing agent, is an anticancer drug that induces cell apoptosis by inhibiting depolymerization of microtubules in between the metaphase and anaphase of the cell mitosis. Recently, its effectiveness and mechanism on various tumor has been reported. However, not much research has been done on the application of Taxol to oral squamous cell carcinoma. Cyclosporin A, which is an immunosuppressant, is being used on cancers and when co-administered with Taxol, effectiveness of Taxol is enhanced by inhibition of Taxol induced multidrug resistance. In this study, Cyclosporin A with different concentration of Taxol was co-administered to HN22, the oral squamous cell carcinomacell line. To observe the cell apoptosis and the mechanisms that take part in this process, mortality evaluation of tumor cell using wortmannin, c-DNA microarray, RT-PCR analysis, cytometry analysis and western blotting were used, and based upon the observation on the effect and mechanism of the agent, the following results were obtained: 1. The HN22 cell line viability was lowest when $100{\mu}M$ of Wortmannin and $5{\mu}g/ml$ of Taxol were co-administered, showing that Taxol participates in P13K-AKT1 pathway. 2. In c-DNA microarray, where $1{\mu}g/ml$ of cyclosporine A and 3mg/ml of Taxol were co-administered, no up regulation of AKT1, PTEN and BAD c-DNA that participate in cell apoptosis was observed. 3. When $1{\mu}g/ml$ of Cyclosporin A was applied alone to HN22 cell line, no difference was found in AKT1, PTEN and BAD mRNA expression. 4. Increased AKT1, mRNA expression was observed when $3{\mu}g/ml$ of Taxol was applied alone to HN22 cell line. 5. When $1{\mu}g/ml$ of Cyclosporin A and Taxol($3{\mu}g/ml\;and\;5{\mu}g/ml$) were co-administered to HN22 cell line, PTEN mRNA expression increased, whereas AKT1 and BAD mRNA decreased. 6. As a result of cytometry analysis, in the group of Cyclosporin A($1{\mu}g/ml$) and Taxol($3{\mu}g/ml$) co-administration, increased Annxin V was observed, which shows that apoptosis occurred by deformation of plasma membrane. However, no significant difference was observed with vary ing concentration. 7. In western blot analysis, no caspase 3 was observed in the group of Cyclosporin A($1{\mu}g/ml$) and Taxol($3{\mu}g/ml$) co-administration. From the results of this study, it can be concluded that synergistic effect can be observed in combination therapy of Taxol and Cyclosporin A on oral squamous cell carcinoma cell line, where decreased activity of the cell line was observed. This resulted in decreased AKT1 and BAD mRNA and increased PTEN mRNA expression and when wortmannin and Taxol were co-administered, the viability decreased which confirms that Taxol decreases the viability of tumor cell line. Hence, when Taxol and cyclosporine A are co-administered, it can be assumed that cell apoptosis occurs through AKt1 pathway.

Phospholipase C-γ Activation by Direct Interaction with β-Tubulin Isotypes (베타 튜불린에 의한 포스포리파제 C-감마1의 활성화)

  • Lee, In-Bum;Kim, Sung-Kuk;Choi, Jang-Hyun;Suh, Pann-Ghill;Chang, Jong-Soo
    • Journal of Life Science
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    • v.16 no.4
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    • pp.612-617
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    • 2006
  • Phosphoinositide-specific phospholipase $C-{\gamma}\;1\; (PLC-{\gamma}\;1)$ has pivotal roles in cellular signaling by producing second messengers, inositol 1,4,5-trisphosphate $(IP_3)$ and diacylglycerol (DG). Tubulin is a main component of microtubules and mitotic spindle fibers, which are composed of ${\alpha}-$ and ${\beta}-tubulin$ heterodimers in all eukaryotic cells. In humans, six ${\beta}-tubulin$ isotypes have been identified which display a distinct pattern of tissue expression. Previously we found that $PLC-{\gamma}\;1$ and one of four ${\beta}-tubulin$ isotypes including ${\beta}1$, ${\beta}2$, ${\beta}3$ and ${\beta}6$, colocalized in COS-7 cells and cotranslocated to the plasma membrane to activate $PLC-{\gamma}\;1$ upon agonist stimulation. In the present study, we demonstrate that the remaining two, tubulin ${\beta}4$ and ${\beta}5$, also showed a potential to activate $PLC-{\gamma}\;1$. The phosphatidylinositol 4,5-bisphosphate $(PIP_2)$ hydrolyzing activity of $PLC-{\gamma}\;1$ was substantially increased in the presence of purified ${\beta}4$ and ${\beta}5$ tubulin in vitro, whereas the activity was not promoted by bovine serum albumin, suggesting that tubulin ${\beta}4$ and ${\beta}5$ also activate $PLC-{\gamma}\;1$. Taken together, our results suggest that all the ${\beta}-tubulin$ isotype activates $PLC-{\gamma}\;1$ activity to regulate cellular signaling.

Does the Gut Microbiota Regulate a Cognitive Function? (장내미생물과 인지기능은 서로 연관되어 있는가?)

  • Choi, Jeonghyun;Jin, Yunho;Kim, Joo-Heon;Hong, Yonggeun
    • Journal of Life Science
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    • v.29 no.6
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    • pp.747-753
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    • 2019
  • Cognitive decline is characterized by reduced long-/short-term memory and attention span, and increased depression and anxiety. Such decline is associated with various degenerative brain disorders, especially Alzheimer's disease (AD) and Parkinson's disease (PD). The increases in elderly populations suffering from cognitive decline create social problems and impose economic burdens, and also pose safety threats; all of these problems have been extensively researched over the past several decades. Possible causes of cognitive decline include metabolic and hormone imbalance, infection, medication abuse, and neuronal changes associated with aging. However, no treatment for cognitive decline is available. In neurodegenerative diseases, changes in the gut microbiota and gut metabolites can alter molecular expression and neurobehavioral symptoms. Changes in the gut microbiota affect memory loss in AD via the downregulation of NMDA receptor expression and increased glutamate levels. Furthermore, the use of probiotics resulted in neurological improvement in an AD model. PD and gut microbiota dysbiosis are linked directly. This interrelationship affected the development of constipation, a secondary symptom in PD. In a PD model, the administration of probiotics prevented neuron death by increasing butyrate levels. Dysfunction of the blood-brain barrier (BBB) has been identified in AD and PD. Increased BBB permeability is also associated with gut microbiota dysbiosis, which led to the destruction of microtubules via systemic inflammation. Notably, metabolites of the gut microbiota may trigger either the development or attenuation of neurodegenerative disease. Here, we discuss the correlation between cognitive decline and the gut microbiota.

Brain-expressed X-linked 2 Binds to Kinesin Superfamily Protein 3A (Brain-expressed X-linked (Bex) 2와 heterotrimeric kinesin-2의 KIF3A와의 결합)

  • Kim, Mooseong;Jeong, Young Joo;Park, Sung Woo;Seo, Mi Kyoung;Kim, Sang Jin;Lee, Won Hee;Urm, Sang-Hwa;Lee, Jung Goo;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.32 no.3
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    • pp.189-195
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    • 2022
  • Kinesin-2 comprises two subfamilies of the heterotrimeric or homodimeric motors found in mammalian cells. Heterotrimeric kinesin-2 consists of kinesin superfamily proteins (KIFs) 3A and 3B and kinesin-associated protein 3 (KAP3), which is a molecular motor protein that moves along microtubules. It plays diverse roles in cargo transport, including anterograde trafficking in cilia, and interacts with many different cargoes and proteins, but their binding proteins have not yet been fully identified. In this study, the yeast two-hybrid assay was used to identify the proteins that interact with the cargo-binding domain (CBD) of KIF3A, and an interaction between KIF3A and brain expressed X-linked 2 (Bex2) was found. Bex2 bound to the CBD-containing C-terminal tail region of KIF3A but did not interact with the same region of KIF3B or KIF5A (a motor protein of kinesin-1). KIF3A interacted with another isoform, Bex1, but did not interact with Bex3. In addition, glutathione S-transferase (GST) pull-downs showed that KIF3A specifically interacts with GST-Bex1 and GST-Bex2 but not with GST alone. When co-expressed in HEK-293T cells, Bex2 co-localized with KIF3A and co-immunoprecipitated with KIF3A and KIF3B but not KIF5B. In combination, these results suggest that Bex2 is capable of binding to heterotrimeric kinesin-2 and may serve as an adaptor protein that links heterotrimeric kinesin-2 with cargo.