• Reproductive and Developmental Biology
• /
• 제34권2호
• /
• pp.73-79
• /
• 2010

We investigated the microtubule dynamics, including the inheritance of donor centrosomes and the mitotic spindle assembly occurring during the first mitosis of somatic cell nuclear transfer (SCNT) embryos in pigs. SCNT embryos were fixed 15 min and 1 h after fusion in order to assess the inheritance pattern of the donor centrosome. The distribution and dynamic of the centrosome and microtubule during the first mitotic phase of SCNT embryos were also evaluated. The frequency of embryos evidencing $\gamma$-tubulin spots (centrosome) was 93.2% in the SCNT embryos 15 min after fusion. In the majority of the SCNT embryos (61.5%), however, no centrosome was observed 1 h after fusion. The frequency of the embryos with no or abnormal mitotic spindles 20 h after fusion was 19.6%. The $\gamma$-tubulin spots were detected near the nuclei of somatic cells regardless of cell cycle phase, whereas $\gamma$-tubulin spots in the SCNT embryos were observed only during the inter-anaphase transition. These results showed that the donor centrosome is inherited into the SCNT embryos, but failed to assemble the normal mitotic spindles during first mitotic phase in some SCNT embryos.

• 한국작물학회지
• /
• 제2권
• /
• pp.3-9
• /
• 1964

소맥, 대두, 면화 및 대맥의 근단에 대하여 저온($0^{\circ}C$~2$^{\circ}C$에 24 또는 12시간) 8-Hydroxyquinolin (0.03과 0.1%에서 2시간 또는 10시간) 및 Colchicine(0.2%와 1.0%에서 2 또는 10시간) 처리를 하고 mitotic cell의 수와 이에 대한 meitaphase의 출현빈도를 검토하였다. 1 저온처리는 모든 작물에 대하여 mitotic cell의 수와 metaphase의 출현빈도를 현저히 증가시켰다. 저온의 정도와 처리시간은 작위에 따라 달라 본시험에서는 소맥을 제외한 타작물에서는 최적온도와 시간은 시험된 시간보다도 짧은 범위 이내에 있는 것으로 생각된다. 2. 8-Hydroxyquinolin의 효과도 작물에 따라 다르나 0.03% 2시간 정도의 처리에서는 대체로 metaphase의 빈도를 증가시키나 mitotic cell의 수를 감소시키는 경향이 있다. 3. Colchicine은 mitotic cell의 증가 및 metaphase 빈도 증가에 항상 유효하였으며 이용에 있어서는 타처리에 비하여 같거나 우수하였다. 4. 본 시험에서의 각종 처리는 염색체를 수축시켰는데 그 정도가 가장 심한 것은 Colchicine이었고 그 다음이 8-Hydroxyquinolin 및 저온처리의 순이었다. 5. 재료에 따라 적당한 온도와 처리시간이 검토된다면 저온처리가 유효하게 적용될 수 있는 가능성이 논의되었다.

• Asian-Australasian Journal of Animal Sciences
• /
• 제2권1호
• /
• pp.35-38
• /
• 1989

Giemsa C-banded mitotic chromosome prepatations from White Leghorn, New Hampshire and Rhode Island Red inbred lines were compared for frequency of C-band regions on individual chromosomes. Except for autosomes 3, 6, 8 and 9 and W sex chromosomes, C-banding was extremely variable in other macrochromosomes. No divergence for C-band difference between homologous chromosomes of these lines was detected. Approximately 75% of the mitotic metaphase microchromosomes have recognizable C-band regions with the current technique.

• Fisheries and Aquatic Sciences
• /
• 제14권4호
• /
• pp.429-433
• /
• 2011

Eggs of Korean rose bitterling Rhodeus uyekii were collected and fertilized to observe temperature-related cleavage rates and mitotic intervals (${\tau}_0$). As the water temperature was increased, the slope of first cleavage frequency with elapsed time after fertilization increased, and approximately 30% of fertilized eggs reached first cleavage frequency at every 15 min. At higher temperatures, eggs developed faster and underwent further identical developmental processes. There were strong, negative correlations between ${\tau}_0$ and water temperatures at all temperatures studied (Y = -1.225X + 70.05, $r^2=0.988$, where Y is ${\tau}_0$ and X is temperature).

• 한국환경성돌연변이발암원학회지
• /
• 제15권2호
• /
• pp.100-105
• /
• 1995

The effects of DNA polymerase $\beta$ on the somatic chromosome mutations and mitotic recombinations were investigated using the transgenic Drosophila beating chimetic gene consisting of a promoter region of Drosophila actin 5C gene and rat DNA polymerase $\beta$. For detecting the somatic chromosome mutations and mitotic recombinations, the heterozygous (mwh/+) strains possessing or lacking transgene poi 13 were used. The spontaneous frequency of small mwh spots, due to deletion or nondisjunction etc., in the non-transgenic w strain and the transgenic p[pol $\beta$]-130 strain was 0.351 and 0.606, respectively. The spontaneous frequency (0.063) of large mwh spots, arises mostly from somatic recombination between the centromere and the locus mwh, in the transgenic p[pol $\beta$]-130 strain was about three times higher than that (0.021) of the non-transgenic w strain. The mutant clone frequencies of small and large mwh spots induced by N-methyl-N'-nitro-N-nitrosoguanidine and ethyl methanesulfonate in the transformant p[pol $\beta$]-130 were higher than those in the host strain w. The present results suggest that rat DNA polymerase $\beta$ participate at least in the somatic chromosome mutations and mitotic recombination processes.

• 미생물학회지
• /
• 제26권2호
• /
• pp.82-87
• /
• 1988

The genetic analysis and characterization of protoplast fusion hybrids between complementary auxotrophic mutants of Candida pseudotropicalis were carried out. Nuclear fusion appeared to occur in fusion hybrids (e.g., F15 and F33), as strongly suggested by isolation of recombinants after mitotic segregation of parental genetic markers. This was confirmed by KNA content, nuclear staining and comparison of survival rate to UV light. After keeping fusion hybrids for approximately one year, the frequency of spontaneous mitotic segregation was $3.0\times 10^{-4}$ - $8.1\times 10^{-4}$ while that of induced mitotic segregation was $1.4\times 10^{-3}$- $1.7\times 10^{-3}$. These results suggested that they maintained stable karyogamy state. It was also found that the production of $\beta$-D-galactosidase from F15, F33 and F158 was somewhat increased when compared with that from either auxotrophic parents or wild type.

• Journal of Plant Biology
• /
• 제38권1호
• /
• pp.33-38
• /
• 1995

Hydroxyurea (HU), a DNA synthesis inhibitor, was tested as synchronizing agent in root-tip meristem of Allium wakegi. Roots were treated with 2.5mM HU for 14 h to accumulate meristem root-tip cells at the G1/S interface. After release from HU block, the cells re-entered the cell cycle with a high degree of synchrony. Synchronized mitotic frequency of A. wakegi was 22.7%, which was about 3.9 times as high as that of the control. The highest metaphase index(23.0%) was obtained when, 6 h after release from the HU block, the roots were treated with 0.05% colchicine for 2 h. Modifying various Giemsa staining protocols defined for animals and a few plant species, G-bands were visualized at prometaphase and metaphase chromosomes of A. wakegi. The higher degree of chromosome condensation, the less differential bands could be resolved. This is the first demonstration introduced partial mitotic synchronization into G-banding in plant.

• Reproductive and Developmental Biology
• /
• 제33권1호
• /
• pp.13-18
• /
• 2009

The aim of this study was to examine the microtubule distributions of somatic cell nuclear transfer (SCNT) and parthenogenetic porcine embryos. Porcine SCNT embryos were produced by fusion of serum-starved fetal fibroblast cells with enucleated oocytes. Reconstituted and mature oocytes were activated by electric pulses combined with 6-dimethlyaminopurine treatment. SCNT and parthenogenetic embryos were cultured in vitro for 6 days. Microtubule assembly of embryos was examined by confocal microscopy 1 hr and 20 hr after fusion or activation, respectively. The proportions of embryos developed to the blastocyst stage were 25.7% and 30.4% in SCNT and parthenogenetic embryos, respectively. The frequency of embryos showing $\beta$-tubulins was 81.8% in parthenogenetic embryos, whereas 31.3% in SCNT embryos 1 hr after activation or fusion. The frequency of the embryos underwent normal mitotic phase was low in SCNT embryos (40.6%) compared to that of parthenogenetic ones (59.7%) 20 hr after fusion or activation (p<0.05). The rate of SCNT embryos with an abnormal mitosis pattern is about twice compared to that of parthenogenetic ones. The spindle assembly and its distribution of SCNT embryos in the first mitotic phase were not different from those of parthenogenetic ones. The result shows that although microtubule distribution of porcine SCNT embryos shortly after fusion is different from parthenogenetic embryos, and the frequency of abnormal mitosis 20 hr after fusion or activation is slightly increased in SCNT embryos, microtubule distributions at the first mitotic phase are similar in both SCNT and parthenogenetic embryos.

• 한국응용곤충학회지
• /
• 제35권4호
• /
• pp.315-320
• /
• 1996

Drosophila의 actin 5C 유전자 promoter에 쥐의 DNA polymerase $\beta$cDNA를 도입시킨 형질전환 초파리가 고감수성 환경성 변이원 검출계로 사용할 수 있는지를 조사하였다. 체세포 염색체 재조환과 체세포 염색체 돌연변이의 검출을 위해서는 geterozygous(mwh/+) 계통을 사용하였다. 염색체상의 결실이나 비분리 등에 의한 small mwh spot의 자연 발생적 빈도는 non-transgenic w 계통과 transgenic p[pol $\beta$]-130 계통에서 각각 0.351 및 0.606 정도였다. 체세포 염색체 재조환에 의한 large mwh spot의 자연 발생적 빈도의 경우는 transgenic p[pol $\beta$]-130 계통(0.063)이 non-transgenic w 계통(0.021)에 비해 약 3배 정도 높게 나타났다. IQ, Glu-P-1 및 {TEX}$AFB_{1}${/TEX} 등의 돌연변이원의 처리에 의한 경우, 두 종류의 mutant clone의 발생 빈도는 쥐의 DNA polymerase $\beta$가 도입된 transgenic p[pol $\beta$]-130 계통이 non-transgenic w 계통에 비하여 모두 약 2-3배 정도 높게 나타났다. 본 연구의 결과는 쥐의 DNA polymerase $\beta$가 최소한 체세포 염색체 돌연변이 유발이나 체세포 염색체 재조환의 생성 과정에 관여함을 의미하며, 형질전환 초파리 계통이 환경성 변이원 검출계로서 충분한 응용가능성이 있음을 보여 주었다.

• 한국동물학회지
• /
• 제14권4호
• /
• pp.175-180
• /
• 1971

5-AU (5-aminouracil)를 처리하므로써 세포분열의 동시화를 촉진시킨 사람의 신장세포의 분열활동, 염색체 이상 및 DNA복제 양상에 미치는 X-선의 영향을 조직배양 및 자기방사법(autoradiography)을 통하여 추구하였다. 5-AU처리구에서 분열활동의 최고점은 5-AU를 처리한 뒤 10시간에서 나타나며, 대조구에 비해서 6배나 높음을 보여준다. 5-AU 처리후 100R의 X-선을 조사한 실험구에서는 X-선의 영향은 주로 세포분열을 지연시키고 분열활동을 저해시킬 뿐 아니라 분열활동의 최고점을 보여주는 시간을 불규칙하게 한다. 대조구에서 세포당 염색체이상율은 0.030에 불과하나 5-AU를 처리할 경우는 0.147로 높아진다. 한편 5-AU+100R 및 5-AU+200R의 X-선 처리구에서 세포당 염색체 이상율은 각각 0.583 및 0.669로 보다 높아짐을 보겠다. 한편 세포당 1R당 평균 염색체 이상율은 0.0035가 된다. 본 실험결과를 통해 보면 5-AU가 표지된 분열상의 출\ulcorner빈도 및 표지강도를 높이고 있음을 알겠는데, 그것은 5-AU가 세포주기중 S기에 놓인 세포를 축적시키는 힘이 있기때문이라고 보겠다. 이와는 반대로 X-선은 세포의 표지강도와 표지된 분열상의 출현빈도를 저하시킨다.