巻号一覧
17 巻 (1992)
- 6 号 p. 335-
- 5 号 p. 257-
- 4 号 p. 209-
- 3 号 p. 151-
- 2 号 p. 87-
- 1 号 p. 1-
17 巻, 2 号
選択された号の論文の9件中1～9を表示しています
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Junji Yamaguchi, Poh-Yam Lim, Kazumi Aratani, Takashi Akazawa1992 年 17 巻 2 号 p. 87-92
発行日: 1992年
公開日: 2006/03/27
ジャーナル フリーA method has been developed to isolate pure preparations of nuclei in high yield from commercially available viable rice embryos (germ), employing extraction with buffer solution containing glycerol (without detergent) and polyamine, followed by centrifugation on a 30% Percoll cushion. The intactness of the isolated nuclei was confirmed by light microscopy as well as electron microscopy. The protein profiles of both whole nuclei and nuclear extracts obtained by SDS-PAGE, organellar marker enzyme activities, DNA and RNA analyses, and in vitro RNA synthesis, all indicate that the highly purified nuclei are isolated from rice embryos.抄録全体を表示PDF形式でダウンロード (1309K) -
Hiroyuki Horiuchi, Masao Hayashi, Haruo Matsuda, Masayoshi Murata1992 年 17 巻 2 号 p. 93-98
発行日: 1992年
公開日: 2006/03/27
ジャーナル フリーCell attachment and spreading of chicken embryo fibroblasts and three mammalian cell lines were promoted by using thrombocyte secretion products (TSP) prepared from chicken thrombocytes which are analogous to mammalian platelets. The cell attachment activity following TSP treatment was interfered with by the addition of a synthetic peptide containing the RGD sequence from the cell attachment region of fibronectin (FN). Cell attachment on TSP-coated plates was inhibited by the addition of anti-chicken FN rabbit antiserum, but not by anti-chicken vitronectin (VN) rabbit antiserum. By immunoblotting, immunofluorescent test and sensitive
Enzyme linked immunosorbent assay using an anti-chicken FN antiserum, the presence of FN in thrombocytes and the release of FN from the cells were demonstrated. The release of FN from thrombocytes was partially induced by the in vitro cultivation of the cells and was further promoted by treatment of the cells with thrombin.抄録全体を表示PDF形式でダウンロード (808K) -
Masaharu Mori, Junjirou Tsuchiyama, Shigeru Okada1992 年 17 巻 2 号 p. 99-104
発行日: 1992年
公開日: 2006/03/27
ジャーナル フリーWe examined the long-term maintenance of multilineal hemopoiesis in a collagen gel culture of mouse bone marrow cells. When cells were inoculated into the gel, stromal cells formed foci that were composed of sinusoidlike capillary structures, fibroblastic cells, adipocytes and macrophages. Many small hemopoietic
foci similar to granulocyte-macrophage colonies (CFU-GM) appeared within a week and disappeared after two weeks. Several large hemopoietic foci appeared after two to three weeks of culture, without a second challenge of marrow cells. These large hemopoietic foci were composed mainly of myeloid cells. Megakaryocytes and mast cells were also observed. When erythropoietin (EPO) was added to the culture at the beginning, the erythroid focus appeared after 3 weeks and the number of megakaryocytes was greater than that in the culture without EPO. However, when EPO was added to the cultures after 6 or 12 weeks, erythroid cells appeared after 1 week and the number of megakaryocytes increased. This hemopoiesis lasted more than 6 months.抄録全体を表示PDF形式でダウンロード (2104K) -
Masamitsu Yamaguchi, Kazufumi Kuroda, Fumiko Hirose, Akio Matsukage1992 年 17 巻 2 号 p. 105-112
発行日: 1992年
公開日: 2006/03/27
ジャーナル フリーAn immunocytochemical method using a specific monoclonal antibody was employed to detect DNA polymerase α in Drosophila melanogaster embryos during the first 13 nuclear division cycles after fertilization. The anti-DNA polymerase a antibody stained the ooplasm of the unfertilized egg, indicating that DNA polymerase a is maternally stored. Strong nuclear staining with the antibody over the weaker staining of the cyto-plasm was observed at interphase throughout the 13 nuclear division cycles. The staining of the cytoplasmic regions surrounding the nucleus was much stronger than the other region of the syncytial cytoplasm until cycle 10. Although prophase nuclei were stained with the antibody, metaphase chromosomes were never stained through-out the 13 cycles. The chromosomal (nuclear) staining reappeared at anaphase until cycle 11 and at telophase in later cycles. The staining of the syncytial cytoplasm except for the cortical region became faint by cycle 13, suggesting the consumption of the maternal storage by this cycle. These results suggest that DNA polymerase a dissociates from chromosomes at the beginning of metaphase; then in later mitotic phases, it is transported from the syncytial cytoplasm into nuclei to participate in formation of the active DNA replication enzyme complex.抄録全体を表示PDF形式でダウンロード (2949K) -
Dai Ayusawa, Sumiko Kaneda, Yoshiyasu Itoh, Hideyo Yasuda, Yasuko Mura ...1992 年 17 巻 2 号 p. 113-122
発行日: 1992年
公開日: 2006/03/27
ジャーナル フリーA temperature-sensitive growth mutant tsFS20 isolated from mouse FM3A cells was identified as a mutant with thermolabile ubiquitin-activating enzyme E1 by transfection with a full-length cDNA encoding the human E1 enzyme and cell-cell hybridization with an authentic E1 mutant ts85 previously isolated from FM3A cells. The resulting transformants produced thermoresistant E1 activity. Upon shift-up of temperature, asynchronously growing tsFS20 cells showed multiple points of cell-cycle arrest. At the nonpermissive temperature, tsFS20 cells that had been synchronized at the G1-S-phase progressed and accumulated in the mid-S-phase,
as evidenced by the absence of G2-specific cdc2 kinase activity, while ts85 mutant cells, the widely used E1 mutant, reached the G2-phase and were arrested. Thus, the E1 mutation seemed to be involved in progression in the S-phase as well as in the G2-phase in the cell cycle. Degradation of short-lived abnormal proteins in tsFS20 cells was decreased to about 50% at the nonpermissive temperature, while the block was fully restored to the wildtype level in the transformant cells. Relevance of the unusually high incidence of the temperature-sensitive E1 mutation was discussed in terms of the E1 as a determinant of heat tolerance of cells.抄録全体を表示PDF形式でダウンロード (1005K) -
Hiroyuki Okamoto, Kazuo Sukegawa, Shunji Tomatsu, Yasuyuki Suzuki, Tad ...1992 年 17 巻 2 号 p. 123-128
発行日: 1992年
公開日: 2006/03/27
ジャーナル フリーTo simplify the process of transfection of human fibroblasts and to acquire a suitable number of transformants, we investigated experimental conditions of electric pulse-induced transfection of human fibroblasts using origin-defective simian virus 40 DNA(SV40 (ori-) DNA). Voltage, pulse duration, number of pulses
and the concentration of SV40 (ori-) DNA led to the formation of 10 to 30 foci/25 cm2 6 weeks after transfection, using 2 to 3 x 106 cells and a square wave pulse generator. Optimal condition was determined to be 2 or 3 pulses at a voltage of 1500 to 2000 V/0.4 cm with 30 μsec pulse width, using 2 μg of linearized SV40(ori-) DNA. With this approach we developed human transformed fibroblasts cell lines with all types of mucopolysaccharidoses. The transformed fibroblasts grew rapidly and the saturation density exceeded that of the parental cells. All the transformed cell clones expressed T antigen, and deficiency in specific enzymes was conserved. A point
mutation which occurred in the human β-glucuronidase gene in a patient with mucopolysaccharidosis type VII was also conserved.抄録全体を表示PDF形式でダウンロード (465K) -
Kenji Sugimoto, Hideyuki Migita, Yoshimasa Hagishita, Hiroaki Yata, Mi ...1992 年 17 巻 2 号 p. 129-138
発行日: 1992年
公開日: 2006/03/27
ジャーナル フリーCentromere protein B (CENP-B) is one of the centromere DNA binding proteins constituting centromere heterochromatin throughout the cell cycles. Some components of mammalian centromeres including CENP-B are target antigens for autoimmune disease patients, often those with scleroderma. Recent isolations of CENP-B genes from human and mouse suggested that CENP-B was highly conserved among mammals. From the previous analysis of the reactivity of patient anticentromere sera, two autoepitopes have been located on the DNA binding domain at the amino-terminal region. The amino acid sequences for both the epitopes are perfectly conserved in the two species, human and mouse. In this study, to identify a human-specific antigenic determinant, the remaining two epitopes were further located in separate carboxyl-terminal regions of human CENP-B. Although the amino acid sequence of one epitope is identical to that of the corresponding region in mouse CENP-B, the other has a less homologous sequence. To confirm that the latter epitope was available for production of human-specific anticentromere antibodies, mice were immunized with the recombinant human CENP-B product. One serum that exclusively stained human centromere structure, but not that of other mammals, was identified in the immunofluorescence microscopic observation. The epitope analysis showed that the less conserved one was recognized by this serum. These results suggested that the corresponding region defines the antigenic determinants for the species specificity.抄録全体を表示PDF形式でダウンロード (1647K) -
Kazue Tsuji, Akemi Ueno, Toshinori Ide1992 年 17 巻 2 号 p. 139-144
発行日: 1992年
公開日: 2006/03/27
ジャーナル フリーGrowth-arrested rat fibroblasts, 3Y1, and human diploid fibroblasts, TIG-1, were induced to synthesize DNA by stimulation with various agents such as fetal bovine serum (FBS), epidermal growth factor (EGF), colcemid, or colchicine. Taxol, a microtubule-stabilizing agent, blocked the induction of DNA synthesis after stimulation with colcemid or colchicine in both cell lines. Taxol inhibited the induction of DNA synthesis after stimulation with FBS or EGF in TIG-1, but did not in 3Y1.12-0-tetradecanoylphorbol-13-acetate (TPA) induced DNA synthesis in TIG-1, which was reduced only partly by taxol. Taxol stabilized or polymerized microtubules in both cell lines. These results indicate that the inhibitory effect of taxol on the induction of DNA syn-thesis varied among cell lines and among growth factors, and suggest that signal transduction processes may be differentiated by taxol sensitivity. In TIG-1 cells, when taxol was added within 6 h, about halfway into the initiation of DNA synthesis after the addition of FBS or EGF, the inhibition of DNA synthesis still occurred. Taxol did not inhibit the induction of c-fos and c-myc genes by FBS or EGF stimulation. Colchicine itself did not in-duce these genes in TIG-1. Thus, taxol appeared to inhibit the induction of DNA synthesis not by blockage in the early transduction process of the growth signal from the cell surface to nuclei but by blockage in processes operating in the mid- or late-prereplicative phase.抄録全体を表示PDF形式でダウンロード (621K) -
Gerhard Czihak, Manabu K. Kojima1992 年 17 巻 2 号 p. 145-150
発行日: 1992年
公開日: 2006/03/27
ジャーナル フリーMitosis of sea urchin eggs was inhibited when exposed to 3μg/ml aphidicolin from the 2-cell stage onwards. Nevertheless the nuclei migrated to the vegetal pole at the time of the fourth unequal division in control eggs. Two or four equal or unequal asters developed. Asters in proximity to the vegetal pole were always considerably smaller than those close to the center of the two blastomeres. In contrast to colchicine, cytokinesis but not migration of the nuclei in the vegetal half was prevented by treatments with 5 μM cytochalasin B or D. Various mitotic figures were formed in the vegetal blastomeres of eggs treated with 0.4 mM colchicine or 3μM griseofulvin after the third cleavage. In some eggs a centrally localized monaster with chromosomes in sphere-like arrangement was formed in others a monopolar mitotic figure pushed the chromosomes in bowl-like arrangements to the most vegetal cortex. In anaphase one set of chromatids migrated to the monopole leaving the scattered sister-chromatids behind. The mechanism of migration of the nuclei and of chromosome arrangement in the metaphase plate is discussed.抄録全体を表示PDF形式でダウンロード (974K)
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