• Title/Summary/Keyword: Cytoplasmic actin

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A Proteomic Screen for Presynaptic Terminal N-type Calcium Channel (CaV2.2) Binding Partners

  • Khanna, Rajesh;Zougman, Alexandre;Stanley, Elise F.
    • BMB Reports
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    • v.40 no.3
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    • pp.302-314
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    • 2007
  • N type calcium channels (CaV2.2) play a key role in the gating of transmitter release at presynaptic nerve terminals. These channels are generally regarded as parts of a multimolecular complex that can modulate their open probability and ensure their location near the vesicle docking and fusion sites. However, the proteins that comprise this component remain poorly characterized. We have carried out the first open screen of presynaptic CaV2.2 complex members by an antibody-mediated capture of the channel from purified rat brain synaptosome lysate followed by mass spectroscopy. 589 unique peptides resulted in a high confidence match of 104 total proteins and 40 synaptosome proteome proteins. This screen identified several known CaV2.2 interacting proteins including syntaxin 1, VAMP, protein phosphatase 2A, $G_{o\alpha}$, G$\beta$ and spectrin and also a number of novel proteins, including clathrin, adaptin, dynamin, dynein, NSF and actin. The unexpected proteins were classified within a number of functional classes that include exocytosis, endocytosis, cytoplasmic matrix, modulators, chaperones, and cell-signaling molecules and this list was contrasted to previous reports that catalogue the synaptosome proteome. The failure to detect any postsynaptic density proteins suggests that the channel itself does not exhibit stable trans-synaptic attachments. Our results suggest that the channel is anchored to a cytoplasmic matrix related to the previously described particle web.

Myoepithelioma Arising From Trachea; A case report (기관의 근상피종 수술치험 1례)

  • 이현석
    • Journal of Chest Surgery
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    • v.26 no.12
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    • pp.962-964
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    • 1993
  • Myoepithelioma of the trachea is an extremely rare disease. The neoplasm shows histologic features identical to those described in myoepithelioma of salivary glands. The myoepithelioma cells demonstrate the numerous myofilament, desmosomal cellular attachment, cytoplasmic glycogen and pinocytotic vesicles. In immunohistochemical study, myoepithelioma cells show the positive antibody reactions to actin, keratin, vimentin and S-100 protein. A case of myoepithelioma arising from trachea in the seventy-two-year old male diagnosed by microscopic finding and immunohistochemistry is presented.

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Ginseng total saponin modulates podocyte p130Cas in diabetic condition

  • Ha, Tae-Sun;Lee, Jin-Seok;Choi, Ji-Young;Park, Hye-Young
    • Journal of Ginseng Research
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    • v.37 no.1
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    • pp.94-99
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    • 2013
  • Proteinuric conditions demonstrate structural and compositional changes of the foot processes and slit diaphragms between podocytes. p130Cas in podocytes serves as an adapter protein anchoring glomerular basement membrane to actin filaments of podocyte cytoskeleton. To investigate the effect of ginseng total saponin (GTS) on the pathologic changes of podocyte p130Cas induced by diabetic conditions, we cultured mouse podocytes under: 1) normal glucose (5 mM, control); 2) high glucose (HG, 30 mM); 3) advanced glycosylation endproducts (AGE)-added; or 4) HG plus AGE-added conditions and treated with GTS. In confocal imaging, p130Cas colocalized with zonula occludens-1 and synaptopodin connecting to F-actin. However, diabetic conditions relocalized p130Cas molecules at perinuclear cytoplasmic area and reduced the intensity of p130Cas. In Western blotting, diabetic conditions, especially HG plus AGE-added condition, decreased cellular p130Cas protein levels at 24 and 48 h. GTS improved such quantitative and qualitative changes. These findings imply that HG and AGE have an influence on the redistribution and amount of p130Cas of podocytes, which can be reversed by GTS.

Regulation of Protein Expression in Mouse Liver by Inorganic Arsenic: Proteomic Analysis (무기비소에 의한 마우스 간의 단백질 발현 조절 : 단백체 분석)

  • Jin Bo-Hwan;Seong Je-Kyung;Ryu Doug-Young
    • Environmental Mutagens and Carcinogens
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    • v.26 no.2
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    • pp.35-40
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    • 2006
  • Background: Inorganic arsenic is a human carcinogen that can target the liver, but its carcinogenic mechanisms are still unknown. Inorganic arsenic induces a spectrum of tumors including hepatocellular carcinoma in mice. Methods: Pregnant C3H mice were supplied with drinking water containing 50 ppm sodium arsenite during their pregnancy. The protein expression profile in the liver of 0.5-day-old. male offsprings exposed transplacentally to sodium arsenite was analyzed using protein 2D gel electrophoresis followed by mass spectrometry (MALDI-TOF). Results: Expression of proteins such as hydroxymethylglutaryl-CoA synthase mitochondrial precursor (HMG-CoA synthase), ${\beta}$-actin (cytoplasmic 1) and apolipoprotein A-IV precursor (Apo-AIV) were induced in mouse liver by sodium arsenite, while uricase (urate oxidase), guanine nucleotidebinding protein beta subunit 2-like 1 (RACK1) and fructose-bisphosphate aldolase B (Aldolase 2) were down-regulated. Summary: Expression of proteins that have been implicated in carcinogenesis, such as HMG-CoA, ${\beta}$-actin, and RACK1, was regulated in the liver of mice transplacentally exposed to inorganic arsenic.

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Puromycin aminonucleoside modulates p130Cas of podocytes

  • Ha, Tae-Sun;Choi, Ji-Young;Park, Hye-Young
    • Clinical and Experimental Pediatrics
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    • v.55 no.10
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    • pp.371-376
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    • 2012
  • Purpose: Puromycin aminonucleoside (PAN) specifically injures podocytes, leading to foot process effacement, actin cytoskeleton disorganization, and abnormal distribution of slit diaphragm proteins. p130Cas is a docking protein connecting F-actin fibers to the glomerular basement membrane (GBM) and adapter proteins in glomerular epithelial cells (GEpCs; podocytes). We investigated the changes in the p130Cas expression level in the PAN-induced pathological changes of podocytes in vitro. Methods: We observed changes in the p130Cas expression in cultured rat GEpCs and mouse podocytes treated with various concentrations of PAN and antioxidants, including probucol, epigallocatechin gallate (EGCG), and vitamin C. The changes in the p130Cas expression level were analyzed using confocal immunofluorescence imaging, Western blotting, and polymerase chain reaction. Results: In the immunofluorescence study, p130Cas showed a diffuse cytoplasmic distribution with accumulation at distinct sites visible as short stripes and colocalized with P-cadherin. The fluorescences of the p130Cas protein were internalized and became granular by PAN administration in a dose-dependent manner, which had been restored by antioxidants, EGCG and vitamin C. PAN also decreased the protein and mRNA expression levels of p130Cas at high doses and in a longer exposed duration, which had been also reversed by antioxidants. Conclusion: These findings suggest that PAN modulates the quantitative and distributional changes of podocyte p130Cas through oxidative stress resulting in podocyte dysfunction.

Effect of Clonorchis sinensis Excretory-secretory Product on the Cultured SD Rat Bile Duct Fibroblast (배양된 흰쥐 담관 섬유모세포에 대한 간흡충 분비배설 물질의 영향)

  • Kwon, Jung-Nam;Min, Byoung-Hoon;Lee, Haeng-Sook;Kim, Soo-Jin;Joo, Kyoung-Hwan
    • Applied Microscopy
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    • v.39 no.2
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    • pp.117-124
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    • 2009
  • Clonorchis sinensis is the most important widely distributed parasite of the human bile duct in East Asia and the most prevalent parasitic helminth in Korea. The prevalence rate of human clonorchiasis has remained at about 2.9% in Korea. C. sinensis induces dilatation of the duct, hyperplasia of the mucosa, metaplasia or neoplasia of the mucosal epithelium, periductal inflammation and fibrosis, and thickening of the ductal wall. Fibroblast are the most common cells in connective tissue and are responsible for the synthesis of extracellular matrix components. The fibrosis associated with chronic inflammation and injury may also contribute to cholangiocarcinoma pathogenesis, particularly through an increase in extracellular matrix components, which participate in the regulation of bile duct differentiation during development. In this study, ultrastructural changes, the distribution of lectin receptors and actin protein in cultured SD rat bile duct fibroblast after infection of C. sinensis were observed. Experimental group had been divided into four groups: normal bile duct fibroblast cultured in basal media (G1); C. sinensis infected bile duct fibroblast cultured in basal media (G2); normal bile duct fibroblast cultured in basal media containing excretory-secretory product (ESP) (G1-1); C. sinensis infected bile duct fibroblast cultured in basal media containing ESP (G2-1). Overall, once a host is infected by C. sinensis, it affects the host to the extent that sialic acid of ductal fibroblast is increased. Number of cytoplasmic process of SD rat bile duct fibroblast was increased. Actin protein and sialic acid were located in cell surface. Fibroblast induced by C. sinensis was not recovered to normal fibroblast. The cytoplasm bulk and cytoplasmic process were increased whereas the growth rate of the fibroblast of infected SD rat was reduced rather than that of normal fibroblast. In result, it inhibits fibroblast proliferation and increases actin protein on fibroblast cytoplasm, and so causes fibroblast metamorphosis and cellular mutation.

Secondary Structure, 1H, 13C and 15N Resonance Assignments and Molecular Interactions of the Dishevelled DIX Domain

  • Capelluto, Daniel G.S.;Overduin, Michael
    • BMB Reports
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    • v.38 no.2
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    • pp.243-247
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    • 2005
  • Dishevelled (Dvl) is a positive regulator of the canonical Wnt signaling pathway, which regulates the levels of $\beta$-catenin. The $\beta$-catenin oncoprotein depends upon the association of Dvl and Axin proteins through their DIX domains, and its accumulation directs the expression of specific developmental-related genes at the nucleus. Here, the $^1H$, $^{13}C$, and $^{15}N$ resonances of the human Dishevelled 2 DIX domain are assigned using heteronuclear nuclear magnetic resonance (NMR) spectroscopy. In addition, helical and extended elements are identified based on the NMR data. The results establish a structural context for characterizing the actin and phospholipid interactions and binding sites of this novel domain, and provide insights into its role in protein localization to stress fibers and cytoplasmic vesicles during Wnt signaling.

Liposarcoma in the lung of a poodle dog

  • Cho, Ho-Seong;Kim, Ok-Jun;Park, Nam-Yong
    • Korean Journal of Veterinary Research
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    • v.46 no.3
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    • pp.263-265
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    • 2006
  • A 15-year-old intact female poodle dog was referred to a local animal clinic showing signs of dyspnea. A radiographic examination revealed multiple nodules in the lung. The following day, the animal died and a necropsy examination revealed multiple nodular masses of varying sizes in the lung. Microscopically, the tumor cells were composed of round to polygonal cells resembling adipocytes with little or no collagenous stroma. Most of the cells contained clear cytoplasmic vacuoles with the nucleus at the periphery while the other cells contained varying numbers of smaller vacuoles. The immunohisto-chemical evaluation yielded a positive reaction to S-100 and vimentin. Negative results were obtained for pancytokeratin, smooth muscle actin, desmin, epithelial membrane antigen and CD68. This case was diagnosed as a well-differentiated liposarcoma.

Imprint Cytology of Hepatic Angiomyolipoma -A Case Report - (간에 발생한 혈관근육지방종의 압착도말 세포 소견 -1예 보고-)

  • Kim, Ae-Ri;Kim, Hong-Jin;Choi, Joon-Hyuk
    • The Korean Journal of Cytopathology
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    • v.19 no.2
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    • pp.188-193
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    • 2008
  • Angiomyolipoma of the liver is a rare benign tumor that's composed of variable mixtures of adipose tissue, smooth muscle and thick-walled blood vessels. We report here on the imprint cytologic features of a hepatic angiomyolipoma in a 47-year-old man. The smears showed spindle and epithelioid tumor cells in clusters, trabeculae and single cells. The spindle cells had elongated, cigar-shaped nuclei with finely granular chromatin and fibrillary cytoplasm. The epithelioid cells had round nuclei with a moderate amount of cytoplasm. Any adipose tissue was not found. Immunohistochemically, both the spindle and epithelioid cells revealed cytoplasmic positivity for smooth muscle actin and HMB-45.

The innate immune response transcription factor Bombyx mori Relish1 induces high-level antimicrobial peptides in silkworm

  • Kim, Seong-Wan;Kim, Seong-Ryul;Goo, Tae-Won;Choi, Kwang-Ho
    • International Journal of Industrial Entomology and Biomaterials
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    • v.37 no.2
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    • pp.49-54
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    • 2018
  • To artificially enhance antimicrobial peptide expression in Bombyx mori, we constructed genetically engineered silkworms overexpressing Rel family transcription factor. The truncated BmRelish1 (BmRelish1t) gene contained a Rel homolog domain (RHD), nuclear localization signal (NLS), acidic and hydrophobic amino acid (AHAA)-rich region, and death domain (DD), but no ankyrin-repeat (ANK) domain. The BmRelish1t gene was controlled by B. mori cytoplasmic actin 3 promoter in the PiggyBac transposon vector. Chromosome analysis of G1 generations of a transgenic silkworm with EGFP expression confirmed stable insertion of BmRelish1t. BmRelish1t gene overexpression in transgenic silkworms resulted in higher mRNA expression levels of B. mori antimicrobial peptides such as lebocin(~20.5-fold), moricin(~8.7-fold), and nuecin(~17.4-fold) than those in normal silkworms.