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Rat basophilic leukemia cells express syntaxin-3 and VAMP-7 in granule membranes

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 2000年第1期271卷 36-41页

作者： Hibi, T Hirashima, N Nakanishi, M Nagoya City Univ Fac Pharmaceut Sci Mizuho Ku Nagoya Aichi 4678603 Japan

In neuronal cells, it is generally agreed that SNARE proteins underlie the release of neurotransmitter. It is controversial, however, whether they also work functionally in the degranulation of RBL-2H3 cells because the expression of SNARE proteins has not been confirmed and the degranulation is not inhibited by tetanus toxin which cleaves one of SNARE proteins, VAMP-2. We investigated the expression and the localization of SNARE proteins including VAMP-7 which is insensitive to tetanus toxin. RT-PCR analysis showed the existence of SNARE proteins, including syntaxin-2, -3, -4, SNAP-23, VAMP-2, and VAMP-7. Experiments using GFP-conjugated proteins revealed that VAMP-7 was localized only in granule membranes, whereas syntaxin-3 was in both the plasma and granule membranes. Upon antigen stimulation, these proteins in granule membranes moved to the cell surface due to the fusion of granules with the plasma membrane. The results suggest the involvement of SNARE proteins in the degranulation of RBL-2H3 cells. (C) 2000 Academic Press.

关键词：氨基酸序列细菌蛋白质类/代谢碱基序列嗜碱粒细胞/代谢细胞膜/代谢细胞质颗粒/代谢细胞质颗粒/超微结构电穿孔绿色荧光蛋白质类细胞内膜/代谢白血病嗜碱粒细胞急性/代谢发光蛋白质类/代谢膜蛋白质类/生物合成膜蛋白质类/代谢显微镜检查共焦分子序列数据质粒 Qa-SNARE蛋白质类 R-SNARE蛋白质类逆转录聚合酶链反应 SNARE蛋白质类肿瘤细胞培养的膜泡运输蛋白质类动物大鼠

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DIFFERENTIAL EFFECTS OF VASOPRESSIN ON WATER, CALCIUM AND LYSOSOMAL ENZYME CONTENTS OF MITOCHONDRIA-RICH AND LYSOSOME-RICH (GRANULAR) EPITHELIAL-CELLS ISOLATED FROM BULLFROG URINARY-BLADDER

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MOLECULAR AND CELLULAR ENDOCRINOLOGY 1976年第2期4卷 89-106页

作者： PIETRAS, RJ NAUJOKAITIS, PJ SZEGO, CM UNIV CALIF DEPT BIOL LOS ANGELES CA 90024 USA UNIV CALIF MOLEC BIOL INST LOS ANGELES CA 90024 USA

Treatment of bullfrog urinary bladder with arginine vasopressin (AVP) elicited a dose-dependent increase in the basal movement of H2O and Na across isolated tissues. Epithelial cells from the mucosal surface and incubated with 10 mU [milliunits] AVP/ml for 30 min retained a greater amount of intracellular H2O and Ca than cells not treated th hormone. Epithelial cells were further separated into 2 major fractions by density gradient centrifugation;cells damaged during these manipulations were separated from viable cells and discarded. Morphological examination of the 2 respective fractions indicated that they largely consisted of mitochondria-rich (MR) and granular (G) cell types which line the lumen of bullfrog bladder. The Ca content of MR cells averaged 25% greater than that of G-type cells. G cells had a higher content of the characteristic lysosomal hydrolases, acid phosphatase and cathepsin B1 than that found in MR cells. Incubation of G cells with AVP elicited significant increments in H2O and Ca contents and extracellular release of lysosomal enzymes as compared to untreated cells. Among MR cells treated with AVP, Cell Ca declines slightly, but no significant increase in H2O content of extracellular hydrolase activity was detected in comparison with paired control cells. The physiological significance of acid proteinase release from G cells treated with AVP was evaluated in experiments with intact bladder. Proteinase inhibitors which suppress the activity of cathepsin B1 selectively antagonized the action of hormone on H2O permeation. Alterations in the Ca and lysosomal hydrolase activity associated with G cells exposed to AVP may contribute to the hormone-induced H2O flow observed across the intact epithelium.

关键词：酸性磷酸酶/代谢无尾目精氨酸加压素/药理学身体水份/代谢钙/代谢组织蛋白酶类/代谢细胞质颗粒/代谢细胞质颗粒/超微结构上皮细胞上皮/药物作用上皮/代谢上皮/超微结构溶酶体/药物作用溶酶体/代谢显微镜检查电子线粒体/药物作用线粒体/代谢线粒体/超微结构牛蛙膀胱/药物作用膀胱/代谢血管升压素类/类似物和衍生物动物

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UNUSUAL CELL STRUCTURES IN TUMOR-LIKE FORMATIONS OF GRACILARIA (RHODOPHYTA)

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ARCHIVES OF MICROBIOLOGY 1976年第2期108卷 167-174页

作者： TRIPODI, G BETH, K UNIV NAPOLI FAC AGRARIA IST BOT I-80055 PORTICI ITALY STAZ ZOOL ANTON DOHRN I-80122 Naples ITALY

This paper deals with EM observations on cultivated plants of the marine red alga G. verrucosa which developed simple galls;also sea collected material, without galls, was studied. The galls showed unusual but characteristic cell structures, caterpillar-like bodies, containing rows of fusiform bodies. These were found mostly in the cytoplasm near the plastids, in one case connected with the endoplasmic reticulum, occasionally even inside the nucleus, and are probably described here for the first time. It does not seem probable that the caterpillar-like bodies represent mitochondria or bacteria, but the hypothesis that fusiform bodies are related to virus-like structures is discussed. The normal tissues as well as the gall tissue of the laboratory plants contained, besides plastids typical for the red algae, another type of plastids characterized by tubular thylakoids.

关键词：藻类红/超微结构细胞质颗粒/超微结构内质网/超微结构显微镜检查电子类器官/超微结构植物病毒

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THROMBOCYTES IN ATHEROSCLEROSIS-RESISTANT AND ATHEROSCLEROSIS-SUSCEPTIBLE BREEDS OF PIGEON - COMPARATIVE ULTRASTRUCTURAL-STUDY

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EXPERIMENTAL AND MOLECULAR PATHOLOGY 1976年第3期25卷 332-343页

作者： LEWIS, JC FUSTER, V KOTTKE, BA MAYO CLIN & MAYO FDN DEPT PATHOL ANAT ROCHESTER MN 55901 USA

Thrombocytes from atherosclerosis-susceptible White Carneau (WC) and atherosclerosis-resistant Show Racer (SR) pigeons were studied using transmission and scanning electron microscopy [SEM]. When fixed in phosphate-buffered glutaraldehyde, thrombocytes from WC and SR are oval to fusiform in shape and have eccentrically placed electron-dense nuclei, large electron-opaque organelles analogous to dense bodies and extensively vacuolated cytoplasm. Ultrastructural studies following latex particle uptake and Ru red staining delineate 2 distinct cytoplasmic vacuole systems. One of the systems incorporates particles from the extracellular medium, and the other is composed of vacuoles containing the electron-dense granules. SEM reveals that SR thrombocytes adhere to and form aggregates on glass. In contrast, thrombocytes from WC pigeons with aortic lesions adhere less and do not form aggregates. Circulating thrombocytes from WC are altered during aortic lesion development.

关键词：动脉硬化/血液动脉硬化/遗传学动脉硬化/病理学血小板/超微结构鸠鸽科细胞质/超微结构细胞质颗粒/超微结构玻璃显微镜检查电子吞噬作用血小板黏附血小板聚集动物

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POLE CELLS OF DROSOPHILA-MELANOGASTER IN CULTURE - NORMAL METABOLISM, ULTRASTRUCTURE, AND FUNCTIONAL CAPABILITIES

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DEVELOPMENTAL BIOLOGY 1979年第2期69卷 451-465页

作者： ALLIS, CD UNDERWOOD, EM CAULTON, JH MAHOWALD, AP INDIANA UNIV DEPT BIOL PROGRAM MOLEC CELLULAR & DEV BIOL BLOOMINGTON IN 47401 USA

The metabolism, ultrastructure and function of mass-isolated pole cells were examined during short-term culture in vitro. In addition to demonstrating that these cells functioned normally in culture, a number of new features of embryonic pole cells were discovered. Cell populations isolated from Renografin density gradients were incubated in medium containing 3H-valine, uridine or thymidine. Although pole cells incorporated similar amounts of valine into protein as other embryonic cells throughout the 1st 6 h in culture, they began to synthesize RNA only after 2 h in culture. Approximately 30% of the pole cells synthesized DNA in vitro and this synthetic activity occurred largely during the 1st h of culture. An ultrastructural analysis of colcemid-treated cells showed that 10% of the pole cells divide shortly after placement in culture. During pole cell culture in vitro, polar granules and nuclear bodies fragment and disperse so that they are eventually not detected in these cells. These changes also occur during pole cell development in vivo. Among the fertile adults which were derived from embryos receiving transplantation of isolated pole cells before and after culture in vitro, 25-30% germ line mosaicism was obtained. These cells are able to follow their normal developmental program in vitro and are able to give rise to functional germ cells in vivo.

关键词：细胞核/超微结构细胞培养的细胞质颗粒/超微结构 DNA/生物合成黑腹果蝇/细胞学黑腹果蝇/胚胎学黑腹果蝇/代谢性腺/胚胎学镶嵌现象蛋白质生物合成 RNA/生物合成动物

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Molecular mechanisms of platelet exocytosis: role of SNAP-23 and syntaxin 2 and 4 in lysosome release

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BLOOD 2000年第5期96卷 1782-1788页

作者： Chen, D Lemons, PP Schraw, T Whiteheart, SW Univ Kentucky Coll Med Dept Biochem Lexington KY 40536 USA

On stimulation by strong agonists, platelets release the contents of 3 storage compartments in 2 apparent waves of exocytosis. The first wave is the release of alpha- and dense core granule contents and the second is the release of lysosomal contents. Using a streptolysin O-permeabilized platelet exocytosis assay, we show that hexosaminidase release is stimulated by either Ca++ or by GTP-gamma-S, This release step retains the same temporal separation from serotonin release as seen in intact platelets. This assay system was also used to dissect the molecular mechanisms of lysosome exocytosis, Lysosome release requires adenosine triphosphate and the general membrane fusion protein, N-ethylmaleimide sensitive factor. Uniquely, 2 syntaxin t-SNAREs, syntaxin 2 and 4, which localize to granules and open canalicular membranes, together with the general target membrane SNAP receptor (t-SNARE) protein SNAP-23 appear to make up the heterodimeric t-SNAREs required for lysosome exocytosis, These studies further show that regardless of stimuli (Ca++ or GTP-gamma-S) serotonin and hexosaminidase release requires the same membrane fusion machinery. (C) 2000 by The American Society of Hematology.

关键词：抗原表面/生理学血小板/药物作用血小板/代谢血小板/超微结构钙/药理学载体蛋白质类/生理学细胞质颗粒/化学细胞质颗粒/超微结构胞吐作用/药物作用胞吐作用/生理学鸟苷5'-O-(3-硫代三磷酸)/药理学溶酶体/药物作用溶酶体/代谢膜蛋白质类/生理学显微镜检查免疫电子神经组织蛋白质类/生理学 Qa-SNARE蛋白质类 Qb-SNARE蛋白质类 Qc-SNARE蛋白质类 SNARE蛋白质类血清素/分泌突触融合蛋白质1 氚/诊断应用膜泡运输蛋白质类人类

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PURIFICATION AND FRACTIONATION OF POTATO AMYLOPLASTS

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ANALYTICAL BIOCHEMISTRY 1975年第1-2期65卷 11-18页

作者： DUFFUS, CM ROSIE, R UNIV EDINBURGH SCH AGRDEPT AGR BIOCHEMEDINBURGH EH9 3JFSCOTLAND

A method is described for the isolation and purification of amyloplasts from mature potato tubers. The low-viscosity, high-density materials Urografin and Ludox TM were used in density gradients to obtain an additional fractionation on the grounds of size. Two main populations were identified. The properties of these materials and the purity of the amyloplast fractions are discussed.

关键词：直链淀粉/分析细胞分级分离/方法离心法梯密度细胞质颗粒/分析细胞质颗粒/超微结构 DNA/分析植物/分析植物/超微结构 RNA/分析

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MAST-CELLS AND MYELOFIBROSIS

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AMERICAN JOURNAL OF CLINICAL PATHOLOGY 1975年第2期63卷 203-209页

作者： UDOJI, WC RAZAVI, SA UNIV CINCINNATI MED CTRDEPT PATHOL234 GOODMAN STCINCINNATIOH 45229

Autopsy of a patient with well-documented myelofibrosis revealed marked proliferation of mast cells associated with areas of bone-marrow and splenic fibrosis. The findings suggest that the local fibrosis represents the healed phase of an inflammatory reaction mediated by mast cells via the release of histamine into the tissue spaces. Tissue mastocytosis may be the pathogenetic mechanism in some cases of myelofibrosis.

关键词：骨髓/病理学细胞质颗粒/超微结构内质网/超微结构肥大细胞/病理学肥大细胞/超微结构原发性骨髓纤维化/血液原发性骨髓纤维化/诊断原发性骨髓纤维化/病理学脾/病理学老年人人类男(雄)性

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BIOCHEMICAL AND MORPHOLOGICAL STUDIES OF RAT SUBMANDIBULAR-GLAND .1. CENTRIFUGAL FRACTIONATION OF GRANULE-RICH FRACTION

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JOURNAL OF DENTAL RESEARCH 1975年第5期54卷 938-947页

作者： HANKS, CT CHAKRABARTI, SG UNIV MICHIGAN SCH DENTINST DENT RESDEPT ORAL PATHOLANN ARBORMI 48104 UNIV MICHIGAN SCH DENTINST DENT RESDEPT ORAL BIOLANN ARBORMI 48104

Submandibular glands from male rats were homogenized in 0.34 M sucrose and 0.5 mM ethylenediaminetetraacetic acid in 10 mM HEPES buffer at a pH of 7.4. The extract was centrifuged and filtered through nuclepore filters to prepare a granule-rich fraction. Electron dense zymogen granules constituted approximately 85% of the particles in this fraction which also contained about a third of the total alkaline esterase activity in the gland.

关键词：酸性磷酸酶/代谢碱类离心法细胞质颗粒/分析细胞质颗粒/酶学细胞质颗粒/超微结构酯酶类/代谢酯类显微镜检查电子肽水解酶类/代谢亚细胞部分/酶学颌下腺/酶学颌下腺/超微结构动物男(雄)性大鼠

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CHARACTERIZATION OF RAT MAST-CELL GRANULE PROTEINS

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ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS 1976年第2期173卷 554-563页

作者： LAGUNOFF, D PRITZL, P UNIV WASHINGTON SCH MED DEPT PATHOL SEATTLE WA 98195 USA

Mast cell granules free of membranes were isolated by differential centrifugation of water-lysed cells. The granules were extracted sequentially with 0.5, 1.0, and 2.0 M KCl. The 0.5 M fraction contained 95% of the N-acetyl-.beta.-glucosaminidase activity;this enzyme probably accounts for no more than 1% of the total granule protein. The 1.0 M fraction contained more than 80% of the granule chymotrypsin-like activity;the chymotrypsin-like enzyme was calculated to represent at least 15% of total granule protein. Heparin was found largely in the 1.0 M extract and in the residue after 2.0 M extraction. The heparin in both fractions had a molecular weight by gel exclusion chromatography considerably in excess of commercial porcine heparin. Acrylamide-gel electrophoresis of granules dissolved in 1% sodium dodecyl sulfate and reduced with dithiothreitol demonstrated 4 major protein bands. The 1.0 M fraction contained the most prominent, rapidly migrating band. The more slowly migrating, higher molecular weight bands appeared in grearer proportion in the 2.0 M and residue fractions. Autodigestion of the 1.0 M extract permitted purification of the mast cell chymotrypsin-like enzyme to specific activities as high as that of crystallized bovine pancreatic .alpha.-chymotrypsin. The mast cell chymotrypsin-like enzyme purified in this way migrated on dodecyl sulfate-gel electrophoresis as a single major band with an estimated molecular weight of 29,000.

关键词：细胞质颗粒/分析细胞质颗粒/超微结构电泳聚丙烯酰氨凝胶肥大细胞/分析肥大细胞/超微结构显微镜检查电子蛋白质类/分析动物男(雄)性大鼠

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