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Y receptor-mediated induction of CD63 transcripts, a tetraspanin determined to be necessary for differentiation of the intestinal epithelial cell line, hBRIE 380i cells

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JOURNAL OF BIOLOGICAL CHEMISTRY 1999年第39期274卷 27914-27924页

作者： Halldén, G Hadi, M Hong, HT Aponte, GW Univ Calif Berkeley Dept Nutr Sci Berkeley CA 94720 USA

Peptide YY (PYY) and neuropeptide Y (NPY) are peptides that coordinate intestinal activities in response to luminal and neuronal signals. In this study, using the rat hybrid small intestinal epithelial cell line, hBRIE 380i cells, we demonstrated that PYY- and NPY-induced rearrangement of actin filaments may be in part through a Y1 alpha and/or a nonneuronal Y2 receptor, which were cloned from both the intestinal mucosa and the hBRIE 380i cells. A number of PYY/NPY-responsive genes were also identified by subtractive hybridization of the hBRIE 380i cells in the presence or absence of a 6-h treatment with PYY. Several of these genes coded for proteins associated with the cell cytoskeleton or extracellular matrix. One of these proteins was the transmembrane-4 superfamily protein CD63, previously shown to associate with beta(1)-integrin and implicated in cell adhesion. CD63 immunoreactivity, using antibody to the extracellular domain, was highest in the differentiated cell clusters of the hBRIE 380i cells. The hBRIE 380i cells transfected with antisense CD63 cDNA lost these differentiated clusters. These studies suggest a new role for NPY and PYY in modulating differentiation through cytoskeletal associated proteins.

关键词：抗原 CD/遗传学抗原 CD/生理学细胞黏附分子/遗传学细胞黏附分子/生理学细胞分化细胞系细胞支架蛋白质类/遗传学细胞骨架/药物作用细胞骨架/生理学细胞骨架/超微结构细胞外基质蛋白质类/遗传学基因表达调控/药物作用杂交细胞肠黏膜小肠动力学神经肽Y/药理学肽YY/药理学血小板膜糖蛋白类/遗传学血小板膜糖蛋白类/生理学 RNA 信使/遗传学受体胃肠激素/生理学受体神经肽Y/生理学转录遗传/药物作用动物大鼠

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Shank, a novel family of postsynaptic density proteins that binds to the NMDA receptor/PSD-95/GKAP complex and cortactin

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NEURON 1999年第3期23卷 569-582页

作者： Naisbitt, S Kim, E Tu, JC Xiao, B Sala, C Valtschanoff, J Weinberg, RJ Worley, PF Sheng, M Massachusetts Gen Hosp Dept Neurobiol Howard Hughes Med Inst Boston MA 02114 USA Harvard Med Sch Boston MA 02114 USA Johns Hopkins Univ Sch Med Dept Neurosci Baltimore MD 21205 USA Univ N Carolina Dept Cell Biol & Anat Chapel Hill NC 27599 USA Pusan Natl Univ Dept Pharmacol Pusan 609735 South Korea

NMDA receptors are linked to intracellular cytoskeletal and signaling molecules via the PSD-95 protein complex. We report a novel family of postsynaptic density (PSD) proteins, termed Shank, that binds via its PDZ domain to the C terminus of PSD-95-associated protein GKAP. A ternary complex of Shank/GKAP/PSD-95 assembles in heterologous cells and can be coimmunoprecipitated from rat brain. Synaptic localization of Shank in neurons is inhibited by a GKAP splice variant that lacks the Shank-binding C terminus. In addition to its PDZ domain, Shank contains a proline-rich region that binds to cortactin and a SAM domain that mediates multimerization. Shank may function as a scaffold protein in the PSD, potentially cross-linking NMDA receptor/PSD-95 complexes and coupling them to regulators of the actin cytoskeleton.

关键词：肌动蛋白类/代谢衔接蛋白质类信号转导 COS细胞载体蛋白质类/分析载体蛋白质类/化学载体蛋白质类/遗传学载体蛋白质类/代谢皮动蛋白细胞骨架/化学细胞骨架/超微结构海马/细胞学细胞内信号肽和蛋白质类膜蛋白质类微丝蛋白质类/分析微丝蛋白质类/代谢显微镜检查免疫电子分子序列数据多基因族神经组织蛋白质类/分析神经组织蛋白质类/代谢神经元/化学神经元/细胞学神经元/代谢蛋白质结构三级受体 N-甲基-D-天冬氨酸/分析受体 N-甲基-D-天冬氨酸/代谢序列同源性氨基酸突触/化学突触/代谢突触/超微结构动物兔大鼠

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Agonist-induced changes in cell shape during regulated secretion in rat pancreatic acini

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JOURNAL OF CELLULAR PHYSIOLOGY 2000年第3期182卷 438-447页

作者： Torgerson, RR McNiven, MA Mayo Clin & Mayo Fdn Dept Biochem & Mol Biol Rochester MN 55905 USA Mayo Clin & Mayo Fdn Ctr Basic Res Digest Dis Rochester MN 55905 USA

The actin cytoskeleton plays an important role in the mediation of exocytosis and the determination of cell shape. Experimentally induced changes in cell shape have been shown to affect stimulated secretion in pancreatic acini. In this study, we have examined whether physiologic agonists induce changes in acinar cell shape to modulate secretion. Computer-enhanced video microscopy, immunofluorescence confocal microscopy, and quantitative Western blotting were used to study cell shape changes and cytoskeletal dynamics in rat pancreatic acini. Amylase assays were performed to study the effect of the actin-myosin cytoskeletal antagonists latrunculin A, BDM, and ML-9 on secretion. We found that pancreatic acini underwent a prominent and reversible shape change in response to the physiologic secretory agonist cholecystokinin. This was accompanied by an apical activation of myosin II as well as a basolateral redistribution of both actin and myosin II. Cytoskeletal antagonists inhibited this shape change and attenuated stimulated amylase secretion. Therefore, in addition to acting as a barrier at the apex, the actin-myosin cytoskeleton may also function to modulate cell shape to further regulate stimulated secretion. (C) 2000 Wiley-Liss, Inc.

关键词：肌动蛋白类/分析肌动蛋白类/代谢淀粉酶类/分泌吖庚因类/药理学二环化合物杂环/药理学细胞大小/药物作用胆囊收缩素/药理学细胞松弛素D/药理学细胞骨架/药物作用细胞骨架/代谢细胞骨架/超微结构二乙酰/类似物和衍生物二乙酰/药理学酶抑制剂/药理学荧光抗体技术显微镜检查共焦显微镜检查电子扫描显微镜检查电视肌球蛋白轻链/拮抗剂和抑制剂肌球蛋白轻链/代谢肌球蛋白类/代谢核酸合成抑制剂/药理学胰腺/药物作用胰腺/分泌胰腺/超微结构大鼠 Sprague-Dawley 噻唑类/药理学噻唑烷类动物男(雄)性大鼠

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AMERICAN BURKITTS LYMPHOMA HAND-MIRROR VARIANT - DETAILED INVESTIGATION OF CYTOLOGIC, ULTRASTRUCTURAL, AND IMMUNOLOGICAL FEATURES

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AMERICAN JOURNAL OF CLINICAL PATHOLOGY 1978年第6期70卷 937-944页

作者： SCHUMACHER, HR RAINEY, T DAVIDSON, L SIMON, D STRONG, M CREEGAN, WJ HOLLOWAY, ML STASS, SA NATL NAVAL MED CTR DEPT LAB MEDHEMATOPATHOL ELECTR MICROSCOPYBETHESDAMD 20014 NATL NAVAL MED CTR DEPT MEDDIV HEMATOL ONCOLBETHESDAMD 20014

Previous studies of patients with lymphoid malignancies have suggested a relationship between survival and the presence of more than 40% hard-mirror cells in the bone marrow. This report represents the first detailed study of a patient with American Burkitt's lymphoma with numerous hand-mirror cells in the bone marrow during the course of the disease. The Burkitt cells were atypical because of their hand-mirror configuration in the bone marrow, the presence of cytoplasmic glycogen, the finding of an IgG surface marker, and a strongly positive acid phosphatase staining reaction. The presence of increased numbers of hand-mirror forms in the bone marrow during the course of the patient's illness may have clinical significance, because they were replaced by non-hand-mirror Burkitt cells in the terminal phase. More reports and investigations of patients with large numbers of hand-mirror forms in the bone marrow are needed to elucidate the full significance of this unusual cellular phenomenon.

关键词：腋骨髓/免疫学骨髓/病理学骨髓/超微结构伯基特淋巴瘤/免疫学伯基特淋巴瘤/病理学伯基特淋巴瘤/超微结构中心粒/超微结构细胞骨架/超微结构免疫球蛋白G/分析免疫球蛋白重链/分析淋巴结/病理学淋巴结/超微结构显微镜检查电子显微镜检查电子扫描显微镜检查相差微管/超微结构线粒体/超微结构多核糖体/超微结构受体抗原 B细胞/分析玫瑰花结形成空泡/超微结构成年人人类男(雄)性

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Pearling in cells: A clue to understanding cell shape

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PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 1999年第18期96卷 10140-10145页

作者： Bar-Ziv, R Tlusty, T Moses, E Safran, SA Bershadsky, A Weizmann Inst Sci Dept Phys Complex Syst IL-76100 Rehovot Israel Weizmann Inst Sci Dept Mat & Interfaces IL-76100 Rehovot Israel Weizmann Inst Sci Dept Mol Cell Biol IL-76100 Rehovot Israel

Gradual disruption of the actin cytoskeleton induces a series of structural shape changes in cells leading to a transformation of cylindrical cell extensions into a periodic chain of "pearls." Quantitative measurements of the pearling instability give a square-root behavior for the wavelength as a function of drug concentration, We present a theory that explains these observations in terms of the interplay between rigidity of the submembranous actin shell and tension that is induced by boundary conditions set by adhesion points. The theory allows estimation of the rigidity and thickness of this supporting shell. The same theoretical considerations explain the shape of nonadherent edges in the general case of untreated cells.

关键词：肌动蛋白类/生理学肌动蛋白类/超微结构细胞系细胞大小细胞骨架/超微结构显微镜检查干涉/方法模型生物学动物

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Membrane skeleton in cultured chick cardiac myocytes revealed by high resolution immunocytochemistry

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HISTOCHEMISTRY AND CELL BIOLOGY 1999年第4期112卷 307-316页

作者： Larsen, TH Dalen, H Sommer, JR Boyle, R Lieberman, M Univ Bergen Expt Cardiol Unit Dept Radiol N-5021 Bergen Norway Duke Univ Med Ctr Dept Cell Biol Div Physiol Durham NC 27710 USA Duke Univ Med Ctr Dept Pathol Durham NC 27710 USA Univ Bergen Dept Pathol Gade Inst N-5021 Bergen Norway

Distribution of cytoskeletal proteins with emphasis on the membrane-cytoskeleton interface was examined in cultured cardiac myocytes. Using specific antibodies recognizing alpha-sarcomeric actin, desmin, beta-tubulin, spectrin/alpha-fodrin and ankyrin, respectively, the cellular localization of these cytoskeletal proteins was detected by laser scanning confocal microscopy. In addition, the fine filamentous structure of these proteins was identified by combining silver-enhanced immunogold labelling with electron microscopy. The latter technique employed the sequence of quick-freezing, deep-etching and rotary shadowing of the specimens. Conventional transmission electron microscopy of the spherical cardiac myocytes revealed a filamentous submembranous layer, approximately 100 nm thick. Specific immunolabelling of alpha-sarcomeric actin and spectrin/alpha-fodrin as well as ankyrin was seen beneath the plasmalemma. A three-dimensional meshwork of spectrin/alpha-fodrin was shown. Numerous desmin filaments that exhibited a tortuous course throughout the cells were also observed running in parallel with the surface in the submembranous area, whereas beta-tubulin was infrequently detected in these areas. In conclusion, the present study shows that spherical cardiac myocytes contain a distinct and complex three-dimensional membrane skeleton. Major constituents of this distinct submembranous layer were spectrin/alpha-fodrin fibres as well as actin and desmin filaments.

关键词：细胞膜/超微结构细胞培养的细胞支架蛋白质类/代谢细胞支架蛋白质类/超微结构细胞骨架/代谢细胞骨架/超微结构细胞内膜/代谢细胞内膜/超微结构显微镜检查共焦显微镜检查免疫电子心肌/细胞学心肌/代谢动物鸡胚

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Adhesion of lymphoid cells to CD44-specific substrata: The consequences of attachment depend on the ligand

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EXPERIMENTAL CELL RESEARCH 2000年第2期256卷 445-453页

作者： Kim, JH Glant, TT Lesley, J Hyman, R Mikecz, K Rush Presbyterian St Lukes Med Ctr Rush Univ Dept Orthoped Surg Sect Biochem & Mol Biol Chicago IL 60612 USA Rush Presbyterian St Lukes Med Ctr Rush Univ Dept Biochem Chicago IL 60612 USA Salk Inst Biol Studies Dept Canc Biol San Diego CA 92186 USA

Interactions between cell-surface adhesion receptors and immobilized specific substrata can exert profound effects on cell morphology. Using phase-contrast microscopy, we show that CD44-expressing mouse lymphoid cells display a spread morphology when adhering to CD44-specific monoclonal antibody (mAb) immobilized on plastic. This spread morphology is different from that of these same cells when adhering to immobilized hyaluronan, the natural ligand of CD44. Morphometric measurements, in combination with intracellular actin staining and fluorescence microscopy, revealed that the adhesion of lymphoid cells to hyaluronan required essentially no cytoskeletal reorganization and resulted in no fundamental change in morphology. On the other hand, cells adhering to immobilized CD44-specific mAb rearranged their actin structure and established multiple membrane contact sites (spread). Cell spreading on antibody, but not attachment to hyaluronan, was inhibited by cytoskeleton-disrupting agents. Transfection of CD44-negative lymphoid cells with full-length and tailless CD44 enabled these cells to bind to both immobilized hyaluronan and mAb. However, the transfectant lacking the cytoplasmic tail of CD44 spread only transiently on the antibody-coated surface. Our results suggest that CD44 may mediate lymphocyte attachment to its carbohydrate ligand hyaluronan by mechanisms broadly similar to those used by selectins. When immobilized CD44-specific antibody is the ligand, however, CD44 may regulate the activity of the cytoskeleton by mechanisms broadly similar to those used by integrins. In the latter case, the cytoplasmic domain of CD44 contributes to cell spreading. (C) 2000 Academic Press.

关键词：肌动蛋白类/代谢抗体/生理学抗原 CD44/免疫学抗原 CD44/代谢抗原 CD44/生理学细胞黏附细胞系细胞骨架/超微结构透明质酸/生理学配体淋巴细胞/代谢淋巴细胞/生理学淋巴细胞/超微结构显微镜检查相差转染动物小鼠

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Systematics of the enigmatic kathablepharids, including EM characterization of the type species, Kathablepharis phoenikoston, and new observations on K-remigera comb. nov.

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PROTIST 1999年第1期150卷 43-59页

作者： Clay, B Kugrens, P Colorado State Univ Dept Biol Ft Collins CO 80523 USA

The colorless flagellate Kathablepharis has consisted of five species based on light microscopic studies, and the ultrastructure of the type species, Kathablepharis phoenikoston, is described for the first time. The heterotrophic, marine flagellate Leucocryptos consisted of two species, but additional ultrastructural details for one of these, Kathablepharis remigera comb, nov. (= Leucocryptos remigera Vors), indicates that it should be transferred to Kathablepharis. The cellular structure of these two species is similar to previously studied kathablepharids. However, there is variation in the feeding apparatus and cytoskeleton. The feeding apparatus of both species has a cytostome, a cytostomal ring, and cytopharyngeal rings. The cytoskeleton consists of inner microtubular arrays and outer or sub-pellicular microtubular arrays. in addition, several features of the flagellar apparatus are described for K. phoenikoston and K. remigera. The ultrastructure of these two species is compared with other kathablepharids to evaluate their taxonomy and phylogeny. We classify Kathablepharis and Leucocryptos in the family Kathablepharididae incertae sedis.

关键词：细胞骨架/超微结构进化摄食行为鞭毛/超微结构显微镜检查电子扫描原生动物/细胞学原生动物/超微结构动物

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Helicobacter pylori induces formation of stress fibers and membrane ruffles in AGS cells by rac activation

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BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 2000年第1期269卷 247-253页

作者： Palovuori, R Perttu, A Yan, Y Karttunen, R Eskelinen, S Karttunen, TJ Oulu Univ Dept Pathol FIN-90401 Oulu Finland Oulu Univ Dept Microbiol FIN-90401 Oulu Finland Oulu Univ Bioctr Oulu FIN-90401 Oulu Finland

Helicobacter pylori induces signaling cascades leading to changes in cytoskeleton and an inflammatory response. Information on the morphological changes and cytoskeletal rearrangements induced by attachment of the bacterium is contradictory and signal transduction pathways are not well known. Since rho family of small GTPases is known to mediate cytoskeletal response to various extracellular stimuli, and is also involved in several other important signal transduction pathways, we have investigated the role of rac and cdc42 in H. pylori-induced cytoskeletal changes in cultured carcinoma AGS cells. AGS cells grown with serum expressed actin filaments in the form of short stress fibers and thin network at the edges, which were depolymerized by removal of serum. In serum-starved cells both type I and type II strains of H. pylori induced formation of actin filaments and lamellipodia-like structures. Microinjection of active rac induced similar changes, but injection of inactive rac prevented the effects of H. pylori, while active or inactive cdc42 did not have any significant effect. Cytoskeletal effects of H. pylori were inhibited by actinomycin D, but not completely by cycloheximide. These results indicate that rac activation is involved in signal transduction cascade leading to cytoskeletal reorganization induced by H. pylori and that gene activation and synthesis of new proteins is necessary in this process. (C) 2000 Academic Press.

关键词：肌动蛋白类/代谢细菌黏附/生理学放线菌酮/药理学细胞骨架/药物作用细胞骨架/代谢细胞骨架/超微结构更生霉素/药理学螺杆菌幽门/致病力蛋白质合成抑制药/药理学信号传导胃肿瘤/代谢胃肿瘤/微生物学胃肿瘤/病理学肿瘤细胞培养的 cdc42GTP结合蛋白质/代谢 racGTP结合蛋白质类/代谢人类

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Tropomyosin isoform 5b is expressed in human erythrocytes: implications of tropomodulin-TM5 or tropomodulin-TM5b complexes in the protofilament and hexagonal organization of membrane skeletons

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BLOOD 2000年第4期95卷 1473-1480页

作者： Sung, LA Gao, KM Yee, LJ Temm-Grove, CJ Helfman, DM Lin, JJC Mehrpouryan, M Univ Calif San Diego Dept Bioengn La Jolla CA 92093 USA Univ Calif San Diego Ctr Genet Mol La Jolla CA 92093 USA Univ Arizona Dept Nutr Sci Tucson AZ 85721 USA Cold Spring Harbor Lab Cold Spring Harbor NY 11724 USA Univ Iowa Dept Biol Sci Iowa City IA 52242 USA

The human erythrocyte membrane skeleton consists of hexagonal lattices with junctional complexes containing F-actin protofilaments of approximately 33-37 nm in length. We hypothesize that complexes formed by tropomodulin, a globular capping protein at the pointed end of actin filaments, and tropomyosin (TM), a rod-like molecule of approximately 33-35 nm, may contribute to the formation of protofilaments, We have previously cloned the human tropomodulin complementary DNA and identified human TM isoform 5 (hTM5), a product of the gamma-TM gene, as one of the major TM isoforms in erythrocytes, We now identify TM5b, a product of the alpha-TM gene, to-be the second major TM isoform, TM5a, the alternatively spliced isoform of the (U-TM gene, which differs by 1 exon and has a weaker actin-binding affinity, however, is not present. TM4, encoded by the delta-TM gene, is not present either. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis, hTM5 comigrated with the slower TM major species in erythrocyte membranes, and hTM5b comigrated with the faster TM major species. TM5b, like TM5, binds strongly to tropomodulin, more so than other TM isoforms, The 2 major TM isoforms, therefore, share several common features: They have 248 residues, are approximately 33-35 nm long, and have high affinities toward F-actin and tropomodulin. These common features may be the key to the mechanism by which protofilaments are formed. Tropomodulin-TM5 or tropomodulin-TM5b complexes may stabilize F-actin in segments of approximately 33-37 nm during erythroid terminal differentiation and may, therefore, function as a molecular ruler. TM5 and TM5b further define the hexagonal geometry of the skeletal network and allow actin-regulatory functions of TMs to be modulated by tropomodulin, (C) 2000 by The American Society of Hematology.

关键词：氨基酸序列载体蛋白质类/血液载体蛋白质类/化学克隆分子细胞骨架/超微结构红细胞膜/超微结构红细胞/代谢红系祖细胞/代谢红系祖细胞/超微结构微丝蛋白质类/血液微丝蛋白质类/化学分子序列数据蛋白质亚型/血液蛋白质亚型/化学蛋白质亚型/遗传学重组蛋白质类/化学重组蛋白质类/代谢序列比对序列同源性氨基酸原肌球调节蛋白质原肌球蛋白/血液原肌球蛋白/化学原肌球蛋白/遗传学动物人类大鼠

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