• 한국발생생물학회:학술대회논문집
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• 한국발생생물학회 2003년도 제3회 국제심포지움 및 학술대회
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• pp.92-92
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• 2003

Accurate analysis of nuclear status is needed when biopsied-blastomeres are used for embryo sexing. In this study, the nuclear status of blastomeres derived from 8- to 16-cell stage IVF bovine embryos was analyzed to evaluate the representative of single blastomere for embryo sexing. When 55 embryos were analyzed by PCR following biopsy, the coincident rate of sex determination between biopsied-single blastomere and matched blastocyst by PCR was 80 %. Karyotyping of biastomeres in 8- 16-cell stage bovine embryos was conducted to assess chromosome status of IVF embryos. To establish karyotyping of blastomeres, concentrations of vinblastine sulfate and duration of exposure time for metaphase plate induction with 8- to 16-cell stage bovine embryos were tested. The most effective condition for induction of metaphase plate (>45%) was 1.0 ug/ml vinblastine sulfate treatment for 15 h. In 22 embryos under the condition, only 8 embryos out of ten that had a normal diploid chromosome complement showed a sex-chromosomal composition of XX or XY (36.4%) and 2 diploid embryos showed mosaicism of the opposite sex of XX and XY in blastomeres of embryo (9.1%). One haploid embryo contained only one X-chromosome (4.5%). Four out of the other 11 embryos having a mixoploid chromosomal complement contained haploid blastomere with wrong sex chromosome (18.2%). These results suggested that morphologically normal bovine embryos derived from IVF had considerable proportion of mixoploid and sex-chromosomal mosaicism which could be the cause of discrepancies of the sex between biopsied-single blastomere and matched blastocyst by PCR analysis.

• 한국약용작물학회지
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• 제18권3호
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• pp.186-190
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• 2010

The chromosome number was identified and fluorescence in situ hybridization(FISH) mapping of 5S and 45S rDNAs were conducted for C. minima and C. lanceolata in the genus Codonopsis from Jeju island. In this study, we have confirmed that the somatic metaphase chromosome number determined as 2n=2x=16 was the same as the findings from the previous studies. While the conventional staining method makes it rather difficult to distinguish satellite chromosomes due to high degree of variability, FISH analysis produced the exact number and location of 5S and 45S rDNAs. Both species in the genus Codonopsis have a pair of 5S rDNA and their gene loci were observed on chromosome 3. Although two pairs of 45S rDNAs (one on chromosome 1 and the other on chromosome 8) were identified in both species, the 45S rDNA signals on chromosome 8 in C. minima were significantly weaker than those on chromosome 1. In addition, the 45S rDNA signals on chromosome 1 in C. lanceolata showed that the chromosome is non-homologus. In this study, we have determined cytogenetic characteristics of C. minima and C. lanceolata according to their gene replication patterns.

• 대한수의학회지
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• 제30권3호
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• pp.317-331
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• 1990

Chromosomes of gonadal tissues from Fasciola spp, Eurytrema pancreaticum and Calicophoron calicophorum occurred Korean cattle were egamined using modified air-drying method. To compare their phenotype with three different genotypes among Fasciola spp, the adult and egg si2e were measured since they have been known as important taxonomical characters. The results obtained were as followed; Cattle liver fluke, Fasciola spp were classified into three types based on their chromosomal complements such as individual with 2o chromosome(diploid), 30 chromosome(triploid) and 20/30 mosaic constitution(mixoploid). The propotions of appearance of three types were 40.00%, 54.29% and 5.71%, respectively. The frequency of three types in type I which was regarded as F gigantica were 58.82% for diploid, 35.29% for triploid and 5.88% for mixoploid, but in type II which was regarded as F hepatica were 72.2% for triploid, 22.22% for diploid and 5.56% for mixoploid. Egg length of triploid forms was significantly larger than that of diploid forms and egg size of mixoploid forms was similiar to that of triploid forms. Worm size of triploid forms was larger than that of diploid forms and was more similar to that of mixoploid forms, but the statistical data were not significant. Diploid chromosome consisted of one pair of metacentric chromosome(m), four pairs of submetacentric chromosomes(sm), five pairs of subtelocentric chromosomes(st), while triploid chromosome consisted of one pair of metacentric chromosome, seven pairs of submetacen.tric chromosomes, one pair of subtelocentric chromosome and telocentric chromosome(t), respectively. In mixoploid chromosome, constitution of the chromosomes of diploid or triploid cell was consistent with that of diploid or triploid. Chromosomes of gonadal tissues from pancreatic fluke, Eurytrema pancreaticum consisted of 13 pairs of homologs(2n=26, n=13). The mitotic and meiotic divisions were observed frequently. In the mitotic metaphase, Karyotype consisted of five pairs of metacentric chromosomes, four pairs of submetacentric chromosomes, three pairs of subtelocentric chromosomes and one pair of telocentric chromosome. Chromosomes of gonadal tissues from stomach fluke, Calicvphoron calicophorum consisted of 9 pairs of homologs(2n=18, n=9). The meiotic divisions was frequently observed, but mitotic divisions was rare. In the mitotic metaphase, Karyotype consisted of two pairs of metacentric chromosomes, three pairs of submetacentric chromosomes and four pairs of subtelocentric chromosomes. Karyotype of Calicophoron calicophorum differed from that of Japanese C calicophorum which was similar to that of Paramphistomum cervi of Korean cattle. Though that of Calicophoron calicophorum of Korean cattle was similar to that of Paramphistomum explanatum of Korean cattle, that have been recognized to be a different species of fluke.

• 한국수정란이식학회지
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• 제22권2호
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• pp.137-141
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• 2007

항산화제는 산소의 저장고로서 무혈청 배양액에서 주요한 작용을 하며, 복합배지에서 유용한 첨가제로 알려져 있다. 따라서 본 연구는 한우 체외 수정란의 배양에 있어서 항산화제인 L-cysteine의 작용과 수정란의 발달 단계별 염색체의 분석을 통하여 체외 수정란의 배양 체계를 수립하고자 실시하였다. 한우 난포란의 체외 성숙은 0.1% PVA, 0.1 mM L-cysteine 첨가 시 체외 성숙율은 73.4%, 94.6%으로 각각 나타냈으며, 처리간에 유의적인 차이를 보였다(p<0.05). 체외 발달율은 20.3%, 10.0%로 5% FBS+TCM199, 0.1 mM L-cysteine+1% BSA 첨가구에서 각각 나타났으며, 처리간에 유의적인 차이는 보이지 않았다. 배양액의 종류에 따른 염색체 분석 결과는 중기상의 수정란은 18.3%, 12.0%를 보였으며, 분석 가능 수정란 수는 6.1%, 4.0%로 5% FBS+TCM199, 0.1mM L-cysteine 첨가구에서 각각 나타났고, 60, XX 2개, 60XY 1개가 5% FBS+TCM199 처리구에서, 60, XX 2개가 0.1 mM L-cysteine 처리구에서 확인되었고, 처리간에 유의적인 차이가 없었다. 수정란의 발달 단계별 염색체 분석 결과는 $4{\sim}16$세포기는 5% FBS-TCM199 배양액과 0.1 mM L-cysteine을 첨가한 배양액에서 18.3, 12.0%의 염색체 중기상을 확인할 수 있었고, 상실기에서는 43.1, 13.0%의 염색체 중기상을 보였으며, 배반포기의 경우는 94.8, 100.0%의 염색체 중기상을 보여 발달 단계가 진행될수록 염색체 중기 상이 많이 나타났다. 이상의 결과로 항산화제인 L-cysteine은 한우 난포란의 체외 성숙 및 발달에 중요한 인자임을 확인하였다.

• 제어로봇시스템학회논문지
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• 제9권10호
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• pp.844-851
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• 2003

We introduces a development result of semi-automatic karyotyping system using image processing method to improve a long time working of the manual method and 5% error of traditional automatic karyotying system for analyzing karyotying. The karyotyping procedures have many routine tasks such as searching metaphases, taking pictures, developing, editing, etc. There are several automatic karyotyping systems in order to reduce the task in advanced countries. However, they are very expensive, applicable to only human chromosome, and have too many functions to use easily. This paper takes aim at high quality image resolution and development of interface that can adjust brightness and contrast of image on-line. The system can be applied to animal and plants as well as human's chromosome. The system developed in this paper is applied to pig and human. The effectiveness of the system is proved by hospitals in Korea.

• 대한본초학회지
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• 제25권1호
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• pp.1-7
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• 2010

Objectives : We investigated genetic toxicity of HMCO5 in relation to chromosome aberration test on Cultured Chinese Hamster Lung (CHL) in the presence and absence of S-9 mix. Methods : Experimental groups were divided into two groups: with S-9mix (+S) or without S-9 mix (-S). -S group was also divided 2 series by treatment hours (6 hr: 6-S; or 24 hr; 24-S). Each group treated with vehicle only (complete culture medium), HMCO5 (1,250, 2,500, $5,000\;{\mu}g/ml$), and cyclophosphamide monohydrate (CPA) and ethylmethanesulfonate (EMS), respectively. Results : HMC05 did not show any aberrant metaphase. However, there were significant (p < 0.01) aberrant metaphases with CPA in S+ and with EMS in S-. Conclusions : These results indicate that HMC05 formula does not show any toxicity in chromosome aberration test.

• Reproductive and Developmental Biology
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• 제30권4호
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• pp.263-270
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• 2006

The Chinese Hamster Ovarian cells CHO-K1 are one of the most extensively used cells for the evaluation of gene expression and toxicology. However, these cells are frequently used for biomedical research without consideration of their cytogenetic characteristics. Therefore, we carried out to investigate the karyologic profiles, the frequency and type of chromosome aberration, and the distribution of telomeric DNA on chromosomes of the CHO-K1 cells. The GTG banding and fluorescence in situ hybridization on CHO-K1 cells were performed to characterize the karyotype and the distribution of telomeric DNA The present study revealed that the chromosome modal number of CHO-K1 cells was 2n=20; eight chromosomes appeared to be identical with those of the normal Chinese hamster, whereas the remaining 12 chromosomes were shown to be translocated, deleted, inversed, or rearranged from Chinese hamster chromosomes. The telomeric DNA on CHO-K1 chromosomes was intensively distributed at the centromeres rather than the ends of chromosomes. In addition, three chromosomes had interstitial telomeres and one marker chromosome entirely consisted of telomeric DNAs. The frequency and type of chromosome aberrations in CHO-K1 cells were examined. Of the 822 metaphase spreads, 68 (8.3%) cells resulted in chromosome aberrations of which the chromosome breakage was the most frequently occurred.

• 한국약용작물학회지
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• 제12권6호
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• pp.515-518
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• 2004

약용식물로 재배되고 있는 지모를 대상으로 McFISH 기법을 이용하여 45S와 5S rDNA 유전자의 염색체상의 위치를 확인하여 물리지도를 확립하였다. 2쌍의 45S rDNA는 1번 염색체의 단완 말단과 3번 염색체의 동원체 부위에서 관찰되었고, 1번 염색체의 signal이 3번 염색체에서의 signal보다 더 강하게 나타났다. 한 쌍의 5S rDNA signal은 45S rDNA signal과 함께 3번 염색체의 동원체 부위에서 관찰되었다.

• Reproductive and Developmental Biology
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• 제28권2호
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• pp.133-137
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• 2004

Sexing from bovine embryos which were fertilized in vitro implicate a possibility of the sex-controlled cattle production. This study was carried out to investigate the possibility of determining of embryo sex by fluorescence in situ hybridization (FISH) technique. FISH was achieved in in vitro fertilized bovine embryos using a bovine Y-specific DNA probe which constructed from the btDYZ-1 sequences. To evaluate Y-chromosome specificity of the FISH probe, metaphase spreads of whole embryos and lymphocytes were prepared and tested. A male-specific signal was detected on 100% of Y chromosome bearing metaphase specimens. Using the FISH technique with a bovine Y-specific probe, 232 whole embryos of 8 cell- to blastocyst-stage were analyzed. Observing the presence of the Y-probe signal on blastomeres, 102 embryos were predicted as male, and 130 embryos as female. The determining rate of embryo sex by FISH technique was about 93% regardless of embryonic stages. In conclusion, the FISH using a bovine Y-specific DNA probe is an accurate, reliable and quick method for determining the sex of bovine embryos.

• 한국약용작물학회지
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• 제10권2호
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• pp.144-146
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• 2002

약용으로 재배되고 있는 지모의 세포유전학적인 연구인 핵형 분석 결과는 다음과 같다. 지모의 체세포 염색체 수는 2n=22였으며. 외형으로 비교해 보았을 때, 3쌍의 상대적으로 길이가 긴 염색체와 길이가 짧은 8쌍의 염색체로 구별이 되었다. 염색체의 평균 길이는 염색체의 길이는 $1.27-3.80\;{\mu}m$로 관찰되었다. Armatio 비교를 통한 핵형 분석에서는 8쌍의 중부 염색체 (염색체 2, 3, 6, 7, 8, 9, 10 및 11), 2쌍의 차중부 염색체 (염색체 4와 5) 그리고 1쌍의 차단부 염색체 (염색체 1)로 구분되었다.