• Title/Summary/Keyword: cytokinesis

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Inadequacy of application of cytokinesis-blocked cells in fish (Rock fish, Sebastes schlegeli) and fowl(chicken) as biological dosimeter for radiation exposure (방사선 피폭의 생물학적 선량측정에 어류(조피볼락, Sebastes schlegeli) 및 조류(닭)의 세포질분열 차단 세포 적용의 부적절성)

  • Kim, Se-Ra;Kim, Tae-Hwan;Ryu, Si-Yun;Jang, Jong-Sik;An, Mi-Young;Kim, Sung-Ho
    • Korean Journal of Veterinary Research
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    • v.42 no.4
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    • pp.451-457
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    • 2002
  • The purpose of the present experiment was to investigate the micronuclei (MN) frequency in cytokinesis-blocked (CB) cells after various doses of gamma-rays in two species (fish and fowl) and so to contribute to the clarification of the question whether these species are suitable as a target organism in the test system. The frequencies of binucleated cells, and gamma-ray-induced MN in CB cells at several doses were measured in three donors of two species. No binucleated cell was noted in erythrocyte. The peaks of binucleated lymphocyte formation were found at a concentration of 2% phytohaemagglutinin (PHA) and $3{\mu}g/m{\ell}$ cytochalasin B (Cyt-B) in fish at 144 hours after incubation and 2% PHA and $6{\mu}g/m{\ell}$ Cyt-B in fowl at 72 hours after incubation. But the micronucleus counts failed to show any evidence of radiation damage. Measurements performed after irradiation showed a dose-related decrease in the formation of binucleated cells in each of the donors studied. Results indicated that the assays were not suitable for this due to blastization inhibition (binucleation failure) after irradiation. We concluded that the use of CB cell from fish and fowl for detecting the results of mdiation exposure was highly questionable.

The cytokinesis-block micronucleus assay as a biological dosimeter in irradiated lymphocyte : Comparison of the response of mouse and human (임파구의 미세핵 분석법을 이용한 생물학적 방사선 피폭선량 측정 : 마우스와 인체에서의 반응비교)

  • Kim, Sung-ho;Cho, Chul-koo;Kim, Tae-hwan;Yoo, Seong-yul;Koh, Kyoung-hwan;Yun, Hyong-geun;Koh, Joo-hwan;Choi, Soo-yong
    • Korean Journal of Veterinary Research
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    • v.33 no.3
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    • pp.487-492
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    • 1993
  • 정상인 말초혈액임파구 및 C57BL/6마우스 비장임파구에 $^{60}Co{\gamma}-rays$를 in vitro상태에서 조사한 후 500개 또는 1000개의 cytokinesis-blocked(CB) lymphocytes의 미세핵(micronuclei)의 발생빈도를 측정하였다. 방사선조사량에 따라 미세핵의 발생빈도는 증가하였으며 linear-quadratic model로 측정한 결과 선량반응곡선의 식은 인체의 경우 $Y=(0.31{\pm}0.049)D+(0.0022{\pm}0.0002)D^2+13.19$($r^2=1.000$)이었으며, 마우스의 경우 $Y=(1.31{\pm}0.264)D+(0.0015{\pm}0.0006)+8.7$($r^2=0.988$)이었다(Y는 1000개의 CB cell 당 미세핵발생빈도, D는 cGy로 표시되는 조사선량). 인체 말초혈액임파구에 대한 마우스 비장임파구의 상대적 생물학적 효과(relative biological effectiveness)는 미세핵의 발생율이 세포당 0.05~0.8의 범위에서 $1.84{\pm}0.48$이었다. 미세핵분석법은 인체 및 동물의 방사선 피폭시 간편하고 빠른 생물학적 선량측정법으로 사용될 수 있을 것이다.

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Radiation exposure dose in human blood lymphocytes as assessed by the CBMN assay

  • Ryu, Tae Ho;Kim, Jin-Hong;Kim, Jin Kyu
    • Journal of Ecology and Environment
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    • v.37 no.4
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    • pp.195-200
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    • 2014
  • The chances of accidental exposure are augmented as the application of ionizing radiation increases in various fields. Such accidental exposures may occur at nuclear power plants, laboratories, and hospitals. Cytogenetic assays have been used for estimating radiation dose in the situation of the accidents. The micronucleus assay has several advantages over the other cytogenetic methods as it is simple and fast. The present study aimed at investigation of the micronuclei frequencies in cytokinesis-block cells in human blood lymphocytes after ${\gamma}$-irradiation and at establishment of a standard dose response relationship. The samples of peripheral blood were obtained from 6 different donors aged between 24 and 30 years old. The bloods were irradiated in vitro with 0-5 Gy. A linear quadratic dose-response equation was obtained by scoring the micronuclei in binucleated cells; $y=27.87x^2+46.13x+2.08$ ($r^2=0.99$). Irradiation caused a significant decrease in the nuclear division index. Necrotic and apoptotic cells increased in number after irradiation in a dose-dependent manner. In conclusion, the conventional cytokinesis-block micronucleus assay has proven to be the great technique in biological dosimetry. Dose-response calibration curve derived from CMBN assay could be used to estimate the exposure dose during a radiological emergency.

Studies on the Organization of 10-nm Filament Ring in Saccharomyces cerevisiae (Saccharomyces cerevisiae 의 10-nm Filament Ring 의 생성기작에 대한 연구)

  • 김성철;정재욱;김형배
    • Korean Journal of Microbiology
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    • v.30 no.5
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    • pp.333-338
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    • 1992
  • Saccharomyces cerevisiae contains 10-nm tilament ring which lies just under the inner surface of the plasma membrane within the mother-bud neck. Although H)-nm filaments may he involved in cellular morphogenesis. their role and organization are not clear. Here we report the production of antihodies specific for the CDel2 protein hy use of gene fusion techniques. and studies on the organization and function of IO-nm filaments using these antibodies. The CDCl2 protein arc translated through the whtlle cell cycle and present in the cytosol. 'They are polymerized just before bud emergence and unpolymerized alier cytokinesis. and do not have organizational relationship with actin. Thc possible role of 10-nm filaments is the determination of bud emergence site and completion of cytokinesis.

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Mad1p, a Component of the Spindle Assembly Checkpoint in Fission Yeast, Suppresses a Novel Septation-defective Mutant, sun1, in a Cell Division Cycle

  • Kim In G.;Rhee Dong K.;Jeong Jae W.;Kim Seong C.;Won Mi S.;Song Ki W.;Kim Hyong B.
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2002.10a
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    • pp.162-172
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    • 2002
  • Schizosaccharomyces pombe is suited for the study of cytokinesis as it divides by forming a septum in the middle of the cell at the end of mitosis. To enhance our understanding of the cytokinesis, we have carried out a genetic screen for temperature-sensitive S. pombe mutants that show defects in septum formation and cell division. Here we present the isolation and characterization of a new temperature-sensitive mutant, sun1(septum uncontrolled), which undergoes uncontrolled septation during cell division cycle at restrictive temperature $(37^{\circ}C)$. In sun1 mutant, actin ring and septum are positioned at random locations and angles, and nuclear division cycle continues. These observations suggest that the sun] gene product is required for the proper placement of the actin ring as well as precise septation. The sun] mutant is monogenic recessive mutation unlinked to previously known various cdc genes of S. pombe. In a screen for $sunl^+$ gene to complement the sun] mutant, we have cloned a gene, $susl^+$(suppressor of sun1 mutant), that encodes a protein of 689 amino acids. The predicted amino acid sequence of $susl^+$ gene is similar to the human hMadlp and Saccharomyces cerevisiae Mad1p, a component of the spindle checkpoint in eukaryotic cells. The null mutant of $susl^+$ gene grows normally at various temperatures and has the increased sensitivity to anti-microtubule drug, while $susl^+$ mutant shows no sensitivity to microtubule destabilizing drugs. The putative S. pombe Sus1p directly interacts with S. pombe Mad2p in yeast two-hybrid assays. These data suggest that the newly isolated susr gene encodes S. pombe Mad1p and suppresses sun] mutant defective in controlled septation in a cell division cycle.

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Analysis of radiation-induced micronuclei and aneuploidy involving chromosome 1 and 4 by FISH technique (FISH 기법을 이용한 방사선에 의한 소핵과 이수성 분석)

  • Chung, Hai-Won;Kim, Tae-Yon;Cho, Yoon-Hee;Kim, Su-Young;Kang, Chang-Mo;Ha, Sung-Whan
    • Journal of Radiation Protection and Research
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    • v.29 no.4
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    • pp.243-249
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    • 2004
  • The cytokinesis-block micronucleus (CBMN) assay in combination with FISH technique using chromosome-specific centromeric probes for chromosome 1 and 4 was performed in mitogen stimulated human lymphocytes which were exposed to x-radiation to identify different sensitivity of chromosomes to the induction of micronuclei(MN) and aneuploidy by radiation. The frequencies of micronucleated cytokinesis-blocked(MNCB) cells and MN in binucleated lymphocytes(BN) increased with the increase in radiation dose. A significant induction of aneuploidy of chromosome 1 and 4 were found. The frequency of aneuploidy of chromosome 1 and 4 in the control were 9 per 2,000 BN cells and this increased to 47 and 71 following irradiation at a dose of 1 and 2 Gy, respectively. The induction of aneuploidy of chromosome 1 was higher than that of chromosome 4. The frequency of aneuploid BN cells with MN exhibiting positive centromere signal for either chromosome 1 and/or 4 increased in a dose dependent manner, and that for chromosome 1 is higher than that for chromosome 4. Among the total induced MN in irradiated lymphocytes, smaller proportion of MN exhibit centromeric signal of chromosome indicating that radiation-induced MN are mainly originated from chromosomal breakage rather than chromosomal non-disjunction. These results suggest that x-radiation can induce aneuploidy and supports the finding that chromosome vary in their sensitivity to aneuploidy induction by x-irradiation.

THE COMBINATION OF MICRONUCLEUS ASSAY AND FISH TECHNIQUE FOR THE EVALUATION OF GENOTOXICITY OF 1, 2, 4-BENZENETRIOL

  • Chung, Hai-Won;Kang, Su-Jin;Kim, Su-Young
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2001.05a
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    • pp.111-111
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    • 2001
  • The cytokinesis-block micronucleus assay have been emerged as one of the preferred method for assessing chromosome damage. Micronucleus are small, extranuclear bodies that are formed in mitosis from acentric chromosomal fragments or chromosome that are not included in each daughter nuclei.(omitted)

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