http://chineseinput.net/에서 pinyin(병음)방식으로 중국어를 변환할 수 있습니다.
변환된 중국어를 복사하여 사용하시면 됩니다.
Application of the modified handmade cloning technique to pigs
( Eun Ji Lee ),( Kuk Bin Ji ),( Ji Hye Lee ),( Hyun Ju Oh ),( Tae Young Kil ),( Min Kyu Kim ) 한국축산학회(구 한국동물자원과학회) 2021 한국축산학회지 Vol.63 No.2
Although somatic cell nuclear transfer (SCNT) is frequently employed to produce cloned animals in laboratories, this technique is expensive and inefficient. Therefore, the handmade cloning (HMC) technique has been suggested to simplify and advance the cloning process, however, HMC wastes many oocytes and leads to mitochondrial heteroplasmy. To solve these problems, we propose a modified handmade cloning (mHMC) technique that uses simple laboratory equipment, i.e., a Pasteur pipette and an alcohol lamp, applying it to porcine embryo cloning. To validate the application of mHMC to pig cloning, embryos produced through SCNT and mHMC are compared using multiple methods, such as enucleation efficiency, oxidative stress, embryo developmental competence, and gene expression. The results show no significant differences between techniques except in the enucleation efficiency. The 8-cell and 16-cell embryo developmental competence and Oct4 expression levels exhibit significant differences. However, the blastocyst rate is not significantly different between mHMC and SCNT. This study verifies that cloned embryos derived from the two techniques exhibit similar generation and developmental competence. Thus, we suggest that mHMC could replace SCNT for simpler and cheaper porcine cloning.
Eun Ji Lee,Kuk Bin Ji,Kang Sun Park,Kyeong Yeob Kim,Beom Sik Kim,Kyu hyun Kim,Ryeong Eun Kim,Eun Young Kim,Ji Hye Lee,Ju Lan Chun,Min Kyu Kim 한국동물생명공학회(구 한국동물번식학회) 2018 발생공학 국제심포지엄 및 학술대회 Vol.2018 No.06
Somatic cell nuclear transfer(SCNT) using micromanipulator have been used as a traditional cloning technique and applied various researches during the last decades. However the micromanipulator is expensive and relatively longer training period is required to operate it efficiently. Handmade cloning(HMC) is an alternative cloning method in a simplified way compared to the traditional cloning. Here, we suggested modified porcine handmade cloning(mHMC) as a new approach to clone porcine embryos as a substitute of the traditional SCNT. In mHMC, a nucleus was removed by an aspiration method by using a glass pipette, instead of bisection method. In this study, we investigated the efficiency of the enucleation method in mHMC by assessing the developmental competence of embryos in comparison with the traditional SCNT. The efficiency of enucleation was evaluated based on the rate of the accuracy and oocyte survivability. The accuracy of enucleation was lower in mHMC compared to those in SCNT(98.01±0.57 vs. 83.83±2.47), and the rate of survived oocytes was also lower in mHMC(96.50±0.84 vs. 90.10±2.11, respectively). And the developmental competence was assessed. The blastocyst rate was significantly higher in mHMC group(13.53±2.08 vs. 20.48±0.99). The levels of apoptosis and ROS were investigated to evaluate embryo quality. The expression of ROS and apoptosis-related genes showed no difference between groups. And the relative expressions of mRNA of pluripotency genes and reprogramming genes were evaluated. Although DNMT1 and DNMT3α were not differently expressed in two groups, the expression of the one of pluripotent gene, Oct4 was significantly higher in mHMC. In conclusion, based on the comparable results of mMHC and SCNT, the mHMC could be a suitable alternative technique to clone embryos in cost effective way compared to traditional SCNT.
김현국,윤지열,이은영,이창근,박중열,유빈,문희범 대한내과학회 2002 대한내과학회지 Vol.63 No.4
Glycogen storage disease type Ia (GSD-Ia) is an autosomal recessive disorder that has defects in glucose-6-phosphatase (G6Pase) in liver, kidney and intestinal mucosa. The defect leads to inadequate conversion of glucose-6-phospate to glucose in the liver and thus makes affected individuals susceptible to fasting hypoglycemia, hyperuricemia, lactic acidemia and hyperlipidemia. Hyperuricemia has been observed in a considerable number of patients and in some of those, clinical gout has occurred. Inhibited tubular secretion of uric acid due to hyperlacticacidemia and ketonemia, and overproduction of uric acid have been postulated as a mechanism for hyperuricemia in patients with GSD-Ia. A 30-year-old male was admitted with fatigue, foot pain and multiple gouty tophi on knee, ankle, and elbow. GSD-Ia and gout were confirmed by analysis of the G6Pase gene and tophi aspiration respectively. He was treated with allopurinol and uncooked cornstarch. After treatment, foot pain improved and the number and size of tophi were decreased.
Neuronal over-expression of human Alzheimer's disease related genes in canines
Chanuka Kulatunga,Dong Eon Kim,JI Hye Lee,Kuk Bin Ji,Eun Ji Lee,Kyeong Yeob Kim,Beom Sik Kim,Kyu Hyun Kim,Ryeong Eun Kim,Yoon Seok Nam,Min Kyu Kim 한국수정란이식학회 2018 한국수정란이식학회 학술대회 Vol.2018 No.11
The early-onset familial Alzheimer's disease (EOFAD/ FAD), the less common type of Alzheimer's disease (AD) currently affects a vast number of individuals worldwide. This type is being inherited as an autosomal dominant fashion. Missense mutations on Amyloid precursor protein (APP) and Presenilins 1 and 2 (PSEN1 & PSEN2) are known as major genetic factors in FAD. Conversely, missense mutations on microtubule-associated protein tau (MAPT) are also thought to involve. Up to date, several triple-transgenic animal models with muted forms of the human APP, PSENs and MAPT have been reported. Compared to other animals, canines are more emotional and their disease signs can be easily diagnosed. This attempt was to develop a triple transgenic canine model for the AD. We have obtained the coding sequences of APP, PSEN1 and MAPT from Dana-Farber/Harvard Cancer Center DNA resource core at HMS and incorporated several common AD mutations. The transgenic construct is composed of hNSE (ENO2) promoter-driven three AD genes fused together with modified 2A sequences. It was transfected into the canine fetal fibroblasts which were then used to perform somatic cell nuclear transfer (SCNT). The viable transgenic embryos were obtained after in vitro culture and the GFP was detected. In this study, we have successfully produced viable triple transgenic canine cloned embryos using SCNT technique. These transgenic canine embryos will be further developed into canines with FAD. The transgenic canines will be a good candidate in the AD research field.
Regulatory Roles of Oocyte Secreted Factors in Canine Cumulus-oocyte Complexes and Granulosa Cells
Lili Zhuang,Dong Eon Kim,Chi Sun Yun,Kuk bin Ji,Eun Ji Lee,Gyeong Yeob Kim,Ji Hye Lee,Min Kyu Kim 한국수정란이식학회 2017 한국수정란이식학회 학술대회 Vol.2017 No.05
Oocyte is the central factor in the bi-directional communication axis in the ovarian follicles. It controls the cumulus or granulosa cells to perform functions which are beneficial for its own development via secreting paracrine growth factors, including GDF9 and BMP15. The aim of this study was to investigate whether the recombinant GDF9 and BMP15 are able to promote meiotic resumption and cumulus expansion of canine COCs during IVM, as well as to demonstrate the actions of GDF9 and BMP15 in regulating the expression of connexin transcripts in the ovarian granulosa cells. As results, GDF9 and BMP15 significantly improved the meiotic resumption rate and cumulus expansion by activating ERK1/2 signaling. Treatments with GDF9 significantly improved the expression of CyclinB1 but inhibited the expression of Cx43 transcripts. In addition, cumulus expansion genes (MAPK1, Ptgs2, Tnfaip6 and Ptx3) were differentially improved by GDF9 and BMP15. In the ovarian granulosa cells, GDF9 suppressed the expression of Cx43 transcripts by binding ALK4/5/7 receptors and activation Smad2/3 signaling, whereas, BMP15 stimulated the expression of Cx43 transcripts by binding ALK2/3/6 receptors and activating Smad1/5/8 signaling. In conclusion, by regulating functions of granulosa/cumulus cells, oocyte has the potential to enhance the growth and maturation of itself.
Lili Zhuang,Bo Myeong Lee,Junbo Wu,Eun Do Lee,Chi Sun Yun,Kuk Bin Ji,Ji Hye Lee,Ju Lan Chun,Min Kyu Kim 한국수정란이식학회 2016 한국수정란이식학회 학술대회 Vol.2016 No.10
Prolonged communication between oocytes and the surrounding somatic cells is one of the unique reproductive physiology in canine. Paracrine Kit ligand (KITL) signaling is a well-known communication between granulosa cells and the oocyte. KITL is a cytokine growth factor secreted by granulosa cells that signals via the c-kit receptor expressed by oocytes. Paracrine factors, including growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15), exert their effects by binding with the kinase receptors expressed on the granulosa cells. However, the regulations of GDF9 and BMP15 in the canine KITL expression are currently poorly understood. Therefore, we investigated the effects of GDF9 and BMP15 on the expression of KITL in canine ovarian granulosa cells in vitro. In Annexin V assay recombinant GDF9 and BMP15 did not induce apoptosis in the cultured ovarian granulosa cells. When treated, FSH significantly increased KITL expression, and hCG suppressed its expression. When both FSH and hCG were treated, the expression of KITL was affected by GDF9 and BMP15 in dose and time dependent manner in the luteal granulosa cells. GDF9 (10 ng/mL) significantly decreased KITL expression after12 h. BMP15 (10 ng/mL) significantly also decreased KITL expression after 24 h. Western blot and immunochemistry results indicate that GDF9 activated Smad2/3. After blocking ALK 4/5/7 receptors by SB, GDF9 failed to activate Smad2/3, also BMP15 did not activate Smad1/5/8 after blocking ALK 2/3/6 receptors by DM. So GDF9 exerts its effects via using ALK 4/5/7 receptors to activate SMAD2/3 signaling, and BMP15 binds ALK 2/3/6 receptors to activate SMAD1/5/8 signaling. The expression of KITL was not changed by SB or DM treatment. However, the effect of GDF9 and BMP15, which decreased the expression of KITL, was suppressed by SB or DM treatment. In conclusion, this study provides the first evidence that recombinant GDF9 and BMP15 decrease KITL expression in canine ovarian granulosa cells.