• Title/Summary/Keyword: Metaphase analysis

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Application of G- anad C-banding Techniques for Chromosome Analysis Using Testis of Mouse (Mouse의 정소이용 염색체 조사에 G- 및 C-banding법의 적용)

  • Choi, Yung-Hyun;Kwon, Yong-Won;Yun, Hee-Sun;Yoo, Mi-Ae;Lee, Won-Ho
    • Journal of Life Science
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    • v.8 no.1
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    • pp.60-66
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    • 1998
  • The mototic and meiotic chromosomal characteristics of ICR mice were investigated with G-and C-banding techniques. For the puposes, the chromosomal preparations were made with the modified air-drying method of Imal et al. Chromosomal analysis using testis could be observed mitotic as well as meitotic chromosomal behaviors, and the centromeric regions of all chromosomes including X chromosome were strongly stained in C-banded preparations. Nineteen autosomal bivalents and a single uniequal terminally associated X-Y bivalent in normal cells were observed during the late prophase and the metaphase of the meiosis I. The mean frequencies of previously dissociated X-Y chromosomes in the primary apermatocytes of the control group were 7.45%, but the frequencies of X-Y dissociation in the alkylating agents-treated group were about 3-4 times higher than that in the control group. Application of C-banding in meiotic stages could be certainly distinguish between vibalent type and univalents type of sex chromosomes.

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Karyotype Analysis and Physical Mapping of rDNAs in Bupleurum longeradiatum (개시호 (Bupleurum longeradiatum)의 핵형분석과 rDNAs의 Physical Mapping)

  • Koo, Dal-Hoe;Seong, Nak-Sul;Seong, Jong-Suk;Bang, Kyong-Hwan;Bang, Jae-Wook
    • Korean Journal of Medicinal Crop Science
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    • v.11 no.5
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    • pp.402-407
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    • 2003
  • Karyotype analysis and chromosomal localization of 5S and 45S rDNAs using multi-color fluorescence in situ hybridization (McFISH) technique were carried out in Bupleurum longeradiatum. Somatic metaphase chromosome number was 2n=12. Karyotype was composed of three pairs of metacentrics (No.3, 4 and 6) and three pairs of submetacentrics (No. 1, 2 and 5). The length of somatic prometaphase chromosomes ranges from 2.55 to $5.05{\mu}m$ with total length of $18.15\;{\mu}m$. In FISH experiment, one pair of 5S rDNA signals was detected on the pericentromeric region of chromosome 4 and one pair of 45S rDNA signals was detected on the telomeric region of chromosome 2.

Karyotype Analysis and Physical Mapping of rDNAs Using McFISH in Jeffersonia dubia Benth (깽깽이풀의 핵형분석과 McFISH를 이용한 rDNA의 물리지도 작성)

  • Kim, Soo-Young;Choi, Hae-Woon;Koo, Dal-Hoe;Kim, Chan-Soo;Bang, Jae-Wook
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.1
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    • pp.48-51
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    • 2005
  • Karyotype analysis and chromosomal locailization of 45S and 5S rDNAs using McFISH (multi-color fluorescence in situ hybridization) were carried out in Jeffersonia dubia Benth., which is one of medicinal plants belonging to Berberidaceae. The somatic metaphase chromosome number was 2n=2x=12 and the size of chromosomes ranged $1.95{\sim}3.50\;{\mu}m$. The chromosome complement consisted of two pairs of metacentrics (chromosomes 1 and 3), two pairs of submetacentrics (chromosomes 2 and 4) and two pairs of subtelocentrics (chromosomes 5 and 6). In McFISH, one pair of 45S rDNA site was detected on the centromeric region of chromosome 2 and three pairs of 5S rDNA sites were detected on the short arm of chromosomes 4, 5 and 6, respectively.

Cytogenetic Analysis of the Triploid Pacific Abalone, Haliotis discus hannai (북방전복, Haliotis discus hannai 3배체의 세포유전학적 연구)

  • Jee, Young-Ju;Chang, Young-Jin
    • The Korean Journal of Malacology
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    • v.28 no.1
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    • pp.37-43
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    • 2012
  • In this study, we invesgated a cytogenetic analysis of the Pacific triploid abalone, Haliotis discus hannai induced by low temperature treatment. We got a lot of mitotic metaphase chromosome spreads from the triploid and diploid Pacific abalones' hatched larvae (trochophores). The chromosome number of diploid abalone was 2n = 36 and that of triploid abalone was 3n = 54, so the chromosome number of triploid abalone was 1.5 times higher than that of diploid abalone. We developed a modified flow cytometric method for Pacific abalone from the existing methods. We uesd 51 months aged triploid and diploid Pacific abalones' hemolymph to get the DNA contents by flow cytometry. The DNA content of diploid abalone was 1.743 pg/cell and the DNA content of triploid abalone was 1.49 times higher than that of diploid one. It proved that triploid abalone was consisted with two sets of maternal diploid and one set of paternal genome.

Development of Bovine Nuclear Transfer Embryos Using Life-span Extended Donor Cells Transfected with Foreign Gene

  • Hwang, Seongsoo;Choi, Eun Joo;You, Seungkwon;Choi, Yun-Jaie;Min, Kwan-Sik;Yoon, Jong-Taek
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.11
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    • pp.1574-1579
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    • 2006
  • This study was performed to determine the developmental potentials of nuclear transfer (NT) embryos using life-span extended cells transfected with a foreign gene as donor cells. A life-span extended bovine embryonic fibroblast cell line was transfected with an expression vector in which the human type II collagen (BOMAR) and ear fibroblasts were used as a donor cell. Cytogenetic analysis was performed to analyze the chromosomal abnormality of donor cells. The fusion rate of 1.8 kV/cm for $15{\mu}sec$ given twice was significantly higher than that of other groups (p<0.05) and the embryos lysed were significantly higher after 1.8 kV/cm for $20{\mu}sec$ given once compared to other groups (p<0.01). The blastocyst development in the ear cell group was statistically significant compared to both BOMAR groups (p<0.01). Both BOMAR groups cultured more than 40 passages (>40 passages) had a lower number of chromosomes; however, fresh granulosa cell (GC) and BOMAR groups cultured less than 20 passages had normal chromosome numbers. Both >40 passages BOMAR groups had numerous obscure debris in metaphase spreads. The transfected foreign gene was expressed in all BOMAR groups, but not in the GC group. Based on these results, the lower developmental potential of NT embryos using life-span extended donor cells transfected with a foreign gene might be a cause of chromosomal abnormality in donor cells.

Identification of Developmental Related Genes in the Lab Animal (실험동물 발생 유전자의 확인)

  • Lee, Jae-Dal
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.10 no.6
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    • pp.1407-1413
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    • 2009
  • The present study was conducted to gain insights into oocyte maturation and early embryo development, SELDI-TOF-MS was used to find the protein candidates that are specifically or prominently expressed in mouse oocytes at the in vitro matured metaphase II (MIl) and germinal vesicle (GV) stages. By using selected CM10 chip, found 16 candidates which were up-regulated in GV stage oocytes compared with in MIl stage oocytes, molecular weight are 8180 (2 candidates), 10226 (5 candidates), 15767 (5 candidates) and 16770 (4 candidates) Da respectively. And 29 candidates were higher in MIl than in GV stage oocytes, molecular weight are 10832 (3 candidates), 17744(8 candidates), 20122 (3 candidates), 22131 (3 candidates), 24857 (7 candidates) and 33507 (5 candidates) Da, respectively. All (45) candidate (0.2 and 1.0 % error tolerances) were performed real time RT-PCR analysis and further selected 13 more potential candidates.

Lower growth factor expression in follicular fluid undergone in-vitro fertilization

  • Han, Myoung-Seok;Park, Seung-Bin;Park, Bang-Ja
    • Clinical and Experimental Reproductive Medicine
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    • v.38 no.4
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    • pp.210-215
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    • 2011
  • Objective: This study was performed to identify whether growth and differentiation factor-9 (GDF-9) and transforming growth factor-${\beta}1$ (TGF-${\beta}1$) expressions would be lower in the follicular fluid (FF) of those over age 35 who underwent IVF than under age 35. Methods: A total of 24 IVF cycles (20 patients) were included in this study. All of patients were stimulated for IVF by the GnRH short protocol and divided into two groups for analysis, according to their age: <35 group (14 cycles, 11 patients) vs. ${\geq}35$ group (10 cycles, 9 patients). The expression levels of GDF-9 and TGF-${\beta}1$ were determined by western blotting and quantitative enzyme-linked immunosorbent assay. Results: The numbers of retrieved oocytes and metaphase II oocytes were significantly lower in the ${\geq}35$ group. Lower expression of GDF-9 and TGF-${\beta}1$ by western blotting in the ${\geq}35$ group were observed as well. The mean GDF-9 and TGF-${\beta}1$ levels by enzyme-linked immunosorbent assay were lower in the ${\geq}35$ group. The values were $6,850.5{\pm}928.4$ ng/L vs. $3,333.3{\pm}1,089.2$ ng/L of GDF-9 ($p$ <0.05) and $3,844.1{\pm}571.1$ ng/L vs. $2,187.7{\pm}754.0$ ng/L of TGF-${\beta}1$ ($p$ <0.05). A negative correlation between GDF-9 and age was observed (r=-0.546, $p$=0.006). Conclusion: GDF-9 and TGF-${\beta}1$ production from stimulated ovaries during IVF appears to decrease with age.

A cytogenetic study of Astragalus koraiensis Y. N. Lee (정선황기의 세포유전학적 연구)

  • Han, Sang Eun;Kim, Hyun-Hee;Heo, Kweon
    • Korean Journal of Plant Taxonomy
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    • v.43 no.2
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    • pp.139-145
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    • 2013
  • This study was carried out to determine the karyotype and chromosomal localizations of 45S and 5S rDNAs using FISH in Astragalus koraiensis. The somatic metaphase chromosome number of this species was 2n = 16 with basic chromosome number of x = 8. The karyotype of A. koraiensis was consisted of six pairs of median region chromosomes(chromosome 1, 3, 4, 5, 6, 8) and two pairs of submedian chromosomes(chromosome 2, 7). Based on the FISH, one pair of 45S rDNA site was detected on the centromeric region of chromosome 5. Whereas, two pair of 5S sites were detected on the short arm of chromosome 4 and centromeric region of chromosome 7, respectively. These are quite different patterns from A. membranaceus, A. membranaceus var. alpinus, and A. mongholicus. Although A. koraiensis is considered as Korean endemic species, therefore, it should be conducted out comparative FISH study with A. sikokianus and A. bhotanensis which are very similar to A. koraiensis morphologically.

The Effect of 5-FU and Radiation on A549 Cells In Vitro (시험관내의 인체폐암 세포 A549의 방사선 및 5-Fluorouracil에 대한 효과)

  • Lee, Myung-Za;Chun, Ha-Chong;Lee, Won-Young
    • Radiation Oncology Journal
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    • v.7 no.1
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    • pp.1-13
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    • 1989
  • Effects of ionizing radiation alone and combined with chemotherapy on tumor growth and it's clonal specificity monitored by changes in distribution of chromosome number were studies in A549 ceil line originated from human adenocarcinoma of the lung. Radiation (300 rad, 600 rad and 900 rad) were delivered with or without 5-FU. Forty eight hours later, 57.5% of growth inhibition of cell w8s seen in cells treated with 5-FU concentration of $0.4{\mu}g/ml$ for 24hr exposure. Cell survival curves after radiation with and without 5-FU were made. Chromosomal analysis of cells in metaphase in control, and in cells treated with 300 rad of radiation, or $0.4{\mu}g/ml$ of 5-FU treatment, and combined treatment of both were done to examine the changes in ploidy and number of chromosome. Radiation combined with S-FU enhanced growth inhibition of A549 cells. However, no evidence of synergegic effects in growth. inhibition was observed in the cells treated with the combination therapy. Pattern of chromosomal distribution of survived cells were shifted from hyperploidy to hypoploidy by single dose of radiation (300 rad). As radiation dose increased a large number of hypoploidy cells were observed. Following treatment of cells with 5-FU, chomosomal distribution of survived cells were also shifted to hypodiploidy which were seen in cells treated with radiation, The ceil treated with 5-FU and fellowed by radiation within 24 hrs had cell with increased number of hypodiploidy cells. Almost same type of chromosomal changes were reproduced in cells treated with combined treatment with radiation and 5-FU. Minor differences were that cells with fewer number of chromosome were more frequent in cells treated with combined therapy. Further increase in cells of hypoploidy (93%) having 1-10 chromosome were induced by additional radiation. Therefore, the enhanced therapeutic effect of 5-FU combined with radiation of A549 cells appeared to be additive rather than synergistic.

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Downstream Networking of $Zap70$ in Meiotic Cell Cycle of the Mouse Oocytes

  • Kim, Hyun-Jung;Lee, Hyun-Seo;Kim, Eun-Young;Lee, Kyung-Ah
    • Development and Reproduction
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    • v.16 no.1
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    • pp.59-67
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    • 2012
  • Previously, we found that $Zap70$ (Zeta-chain-associated protein kinase) expressed in the mouse oocytes and played significant role in completion of meiosis specifically at MI-MII (metaphase I-II) transition. Microinjection of $Zap70$ dsRNA into the cytoplasm of germinal vesicle oocyte resulted in MI arrest, and exhibited abnormalities in their spindles and chromosome configurations. The purpose of this study was to determine the mechanisms of action of $Zap70$ in oocyte maturation by evaluating downstream signal networking after $Zap70$ RNAi (RNA interference). The probe hybridization and data analysis were used by Affymetrix Gene Chip Mouse Genome 430 2.0 array and GenPlex 3.0 (ISTECH, Korea) software, respectively. Total 1,152 genes were up (n=366) and down (n=786) regulated after $Zap70$ RNAi. Among those genes changed, we confirmed the expressional changes of the genes involved in the regulation of actin cytoskeleton and MAPK (mitogen-activated protein kinase) signaling pathway, since the phenotypes of $Zap70$ RNAi in oocytes were found in the changes in the chromosome separation and spindle structures. We confirmed the changes in gene expression in the actin skeletal system as well as in the MAPK signaling pathway, and concluded that these changes are main cause of the aberrant chromosome arrangement and abnormal spindles after $Zap70$ RNAi.