간행물
Reproductive & developmental biology
- 발행기관 한국동물번식학회
- 자료유형 학술지
- 간기 계간
- ISSN 1738-2432 (Print)2288-0151 (Online)
- 수록기간 1977 ~ 2018
- 주제분류 농수해양 > 축산학 농수해양 분류의 다른 간행물
- 십진분류KDC 527DDC 636
권호리스트/논문검색
Volume 30 No 2 (2006년 6월) 11건
1.
2006.06
구독 인증기관 무료, 개인회원 유료
Hyun Gi Lee, Jin-Ki Park, Sung-Woo Kim, Eun-Mi Ko, Byoung-Ju Kim, Su-Jin Jo, Sung-June Byun, Boh-Suk Yang, Won-Kyong Chang, Hoon Taek Lee, Poongyeon Lee
This study was conducted to examine the effect of IRES controlled reporter gene on screening and production of recombinant human erythropoietin (EPO) proteins from cultured CHO cells. The cDNA was cloned for EPO from human liver cDNA. Using site-directed mutagenesis, we generated recombinant human EPO (rhEPO) with two additional N-glycosylations (Novel erythropoiesis-stimulating protein: NESP). Wild type hEPO and NESP were cloned into expression vectors with GFP reporter gene under regulatory control of CMV promoter and IRES so that the vectors could express both rhEPO and GFP. The expression vectors were transfected to cultured CHO-K1 cells. Under microscopy, expression of GFP was visible. Using supernatant of the culture, ELISA assay, immunocytochemistry and in vitro assay using EPO dependant cell line were performed to estimate biological activity to compare the production characteristics (secretion levels, etc.) between rhEPO and NESP. The activity of NESP protein, obtained by mutagenesis, was described and compared with its rhEPO counterpart produced under same conditions. Although NESP had less secretion level in CHO cell line, the biological activity of NESP was greater than that of rhEPO. These results are consistent with previous researches. We also demonstrated that rhEPO and GFP proteins expressed simultaneously from transfected CHO cell line. Therefore we conclude that use of GFP reporter gene under IRES control could be used to screen and produce rhEPO in cultured CHO cells.
4,000원
2.
2006.06
구독 인증기관 무료, 개인회원 유료
Joo-Hee Han, Sungwoo Kim, Poongyeon Lee, Chun-Gyu Park, Hyun-Gi Lee, Boh-Suk Yang, Ki-Hyeong Rhee, Chang-Hyung Lee, Hoon-Taek Lee, Won-Kyong Chang, Jin-Ki Park
The possibility of producing transgenic embryos expressing the green fluorescence protein (GFP) gene have been evaluated after transfer of exogenous gene into the porcine zygote cytoplasm using the intracytoplasm sperm injection (ICSI) as gene delivery method. For DNA binding to sperm heads, 0.05% Triton X-100 or Lipofectin was used. After injection of the sperm bound to DNA by means of Lipofectin or Triton X-100 triturate, the blastocyst formation rates on day 6 were not significantly different from that of ICSI only group (18.8, 19.2 and 25.3%). In terms of GFP expression, more embryos were in GFP form in Triton X-100 group than in Lipofectin group (40.6 vs 36.4%), while percentage of non-mosaic embryos expressing the GFP gene in all blastomere was higher (P<0.05) in Lipofectin group than in Triton X-100 group (4.2 vs 0.9%). ICSI embryos derived from sperm treated with Lipofectin/DNA complex was transferred into 3 recipients and were collected by uterine flushing on days 5, 7 and 15 after embryo transfer, and then GFP expression was observed by a fluorescence microscopy. Over 26% of the collected embryos were normally expressed GFP gene. These results suggest that foreign gene transfer method with DNA bound sperm caused minimal damage to structure of oocytes that can result to full development of porcine embryos. This was confirmed in this study when the embryos that were transferred after ISCI of DNA bound sperm had a normal development and gene expression until preimplantation.
4,000원
3.
2006.06
구독 인증기관 무료, 개인회원 유료
To study the signaling effect of insulin-like growth factor-Ⅰ(IGF-1), transgenic mice containing IGF-1 Receptor (IGF-1R) cDNA fused to metallothionein promoter were produced by DNA microinjection into the pronucleus of mouse zygote. Three founders were produced with transgenic mice containing IGF-1R gene. Transgenic mice lines contained approximately 4～20 copies of transgenes per cell and transmission of this gene into the progeny with Mendelian manner were determined. The founder mice were mated with normal mice to produce F1 mice and then F2 mice. Transmission rates of IGF-1R transgene in the progeny mice were 25～60% in F1 generation and 40～50% in F2 generation. The mRNA expression of IGF-1R transgene in liver was analyzed using RT-PCR for IGF-1R gene in liver. When body weights of transgenic pups were measured during 4, 10 and 14 weeks after birth, IGF-1R transgenic mice grew faster than non transgenic littermates. This study indicated that growth regulation by IGF-1 signaling through IGF-1R can be elucidated using IGF-1R transgenic mice.
4,000원
4.
2006.06
구독 인증기관 무료, 개인회원 유료
Mohammad-Shamim Hossein, Min-Kyu Kim, Goo Jang, Yuda-Heru Fibrianto, Huyn-Joo Oh, Hye-Jin Kim, Joung-Joo Kim, Sung-Keun Kang, Byeong-Chun Lee
The growing oocytes become progressively capable of resuming meiosis, and full meiotic competence appear when they are about 80% of the size of fully grown oocytes. As hormonal influences vary at different stages of reproductive cycle, the size of oocytes may vary according to the reproductive stages. The present study was designed to compare the diameter between the ovulated and freshly collected immature canine oocytes. The ovulated oocytes were collected 72 hr after ovulation by oviductal tube flushing by laparotomy under general anesthesia. Immature oocytes were collected by ovarian slicing method. Diameter of all oocytes was measured directly using epiflurescence microscope with a calibrated micro-eyepiece micrometer at ×200 magnification. The thickness of zona pellucida and diameter of cytoplasm were measured separately and recorded. A total of 2209 zona intact oocytes were collected, among them 628 from anestrus, 675 from follicular, 838 from luteal and 68 by fallopian tubes flushing methods. The average number of oocytes was 104.7, 168.8, 119.7 and 11.3 for anestrus, follicular, luteal and fallopian tubes flushing methods, respectively. The average diameters of the ooplasm and oocyte were significantly varied in different reproductive stages as well as with ovulated oocytes (P<0.05). The average diameter of ooplasm and oocyte was 115.6 and 127.7, 143.0 and 162.0, 134.6 and 150.6, 159.6 and 185.6 for anestrus, follicular, luteal and ovulated oocytes, respectively. Highest number of oocytes with larger diameter could be collected from the follicular and luteal stages. In conclusion, the follicular and luteal ovaries are the best sources of oocytes for canine IVM.
4,000원
5.
2006.06
구독 인증기관 무료, 개인회원 유료
Development of placenta is a complex process that is critical for the pregnancy and controlled by many factors including cytokines, hormones, growth factors and apoptotic molecules. Recently, it has been shown that progranulin (PGRN) functions in growth of embryo and trophectoderm as well as cell migration. To initiate understanding the role of PGRN in human placental development, we investigated the expression of PGRN mRNA and protein in early and late gestation human placentas, term cytotrophoblast cells and two choriocarcinoma cell lines, JEG-3 and Jar. Reverse transcriptase polymerase chain reaction identified mRNAs derived from the PGRN gene in all samples. Immunoblot analysis showed that PGRN proteins are present in early and late gestation human placentas with decreasing levels over gestation and that PGRN proteins are present in normal and transformed trophoblast cells. Immunohistochemical analysis using paraformaldehyde-fixed tissue sections taken from early and late stages of pregnancy showed that PGRN proteins are present in cytotrophoblast cells, syncytiotrophoblast and extravillous cytotrophoblast cells and that expression pattern of PGRN differed according to the stage of cell differentiation. The results of this study are consistent with the hypothesis that PGRN proteins have critical roles in placental development and suggest that PGRN may function in trophoblast cell growth and differentiation.
4,000원
6.
2006.06
구독 인증기관 무료, 개인회원 유료
Cyclin B1 is known to reflect the M-phase promoting factor (MPF), a universal regulator of G2/M-phase transition, activity during the process of oocytes maturation. To investigate whether culture condition affects the maturation rate and the expression of cyclin B1 protein, bovine immature oocytes are stimulated and cultured according to the following protocols: Experiment 1: denuded oocytes (denude) only, COC only, denuded oocytes + granulosa cells (denude + GCs) and COC + GCs; Experiment 2: no-activation (control), 7% ethanol for 5 min and 10 l/ml ionomycin for 5 min at immediately before maturation. The maturation rates of denude and no-activation group were significantly lower in both experiments (P<0.05), respectively. Co-culture or stimulation method in bovine immature oocytes culture increases the cyclin B1 expression significantly in both experiments (P<0.05). Based on these results, culture condition affects the maturation rate and the expression of cyclin B1 protein during the first meiotic maturation in bovine immature oocytes.
3,000원
7.
2006.06
구독 인증기관 무료, 개인회원 유료
Hai-Feng Jin, B. Mohana Kumar, Sung-Keun Cho, Sun-A Ock, Byeong-Gyun Jeon, S. Balasubramanian, Sang-Yong Choe, Gyu-Jin Rho
4,000원
8.
2006.06
구독 인증기관 무료, 개인회원 유료
Jung-Gon Kim, B. Mohana Kumar, Sung-Keun Cho, Sun-A Ock, Byeong-Gyun Jeon, S. Balasubramanian, Gyu-Jin Rho, Sang-Yong Choe
This study was conducted to detect the apoptosis incidence in blastocysts and to compare the abundance of Bax, Bcl2L1, VEGF and FGFR2 in in vitro fertilized (IVF), parthenogenetic (PAT) and nuclear transfer (NT) embryos. Oocytes matured for 40 hr were enucleated and reconstructed with confluenced fetal fibroblasts (FFs) derived from a ～45 day fetus. Reconstructed eggs were then fused with 2 DC pulses (2.0 kV/cm, 30 μsec) and cultured with 7.5 μg/ml cytochalasin B for 3 hr. Parthenotes (PAT) were produced with the same electric strength and culture for NT eggs. The embryos were cultured in NCSU-23 medium at 39℃, 5% CO2, 5% O2 in air. In 3 runs, set of 10 embryos at the 4-cell to blastocyst stages were used to extract total RNA for analyzing the gene expression patterns of pro-apoptotic (Bax), anti-apoptotic (Bcl2L1), vasculogenesis (VEGF), implantation (FGFR2III) using real-time quantitative PCR. Cleavage and blastocyst rates were significantly higher (P<0.05) in IVF and PAT (79.3±8.5 and 25.5±6.1, and 85.0±6.4 and 38.6±5.5, respectively) than NT counterparts (65.1±5.2 and 15.6±3.0, respectively). Significantly higher (P<0.05) total cells were observed in IVF controls and PAT (34.7±5.8 and 38.1±4.1) than NT embryos (24.8±3.2). Apoptosis index was significantly lower (P<0.05) in IVF than NT embryos. The Relative abundances (RA) of Bax and VEGF were significantly higher (P<0.05) at blastocyst stage in NT than IVF control. The RA of Bcl2L1 and FGFR2III were significantly higher (P<0.05) at blastocyst stage in IVF than NT. The present study observed the abnormal gene expressions in NT embryos at various developmental stages, suggesting certain clues to find out the cause of the low efficiency of NT to term.
4,000원
9.
2006.06
구독 인증기관 무료, 개인회원 유료
4,000원
10.
2006.06
구독 인증기관 무료, 개인회원 유료
Major characteristics of embryonic stem cells (ESCs) are sustaining of stemness and pluripotency by self-renewal. In this report, transcriptional profiles of the molecules in the developmentally important signaling pathways including Wnt, BMP4, TGF-β, RTK, Hh, Notch, and JAK/STAT signaling pathways were investigated to understand the self- renewal of mouse ESCs (mESCs), J1 line, and compared with the NIH3T3 cell line and mouse embryonic fibroblast (MEF) cells as controls. In the Wnt signaling pathway, the expression of Wnt3 was seen widely in mESCs, suggesting that the ligand may be an important regulator for self-renewal in mESCs. In the Hh signaling pathway, the expression of Gli and N-myc were observed extensively in mESCs, whereas the expression levels of in a Shh was low, suggesting that intracellular molecules may be essential for the self-renewal of mESCs. IGF-I, IGF-II, IGF-IR and IGF-IIR of RTK signaling showed a lower expression in mESCs, these molecules related to embryo development may be restrained in mESCs. The expression levels of the Delta and HES5 in Notch signaling were enriched in mESCs. The expression of the molecules related to BMP and JAK-STAT signaling pathways were similar or at a slightly lower level in mESCs compared to those in MEF and NIH3T3 cells. It is suggested that the observed differences in gene expression profiles among the signaling pathways may contribute to the self-renewal and differentiation of mESCs in a signaling-specific manner.
4,600원
11.
2006.06
구독 인증기관·개인회원 무료
