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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.
Chi Sun Yun,Dong Eon Kim,Chanuka Kulatunga,Eun Ji Lee,Ji Hye Lee,Eun Young Kim,Ju Lan Chun,Min Kyu Kim 한국동물생명공학회(구 한국동물번식학회) 2018 발생공학 국제심포지엄 및 학술대회 Vol.2018 No.06
The endemic species of sika deer in Northeast Asia is endangered animals in South Korea. In the 1970s, some subspecies (C.n.taiouanus and C.n.yesoensis) were introduced from other countries for restoration, and were not the endemic species (C.n.hortulorum) of Northeast Asia. However, there has not been a sufficient study about endemic and reintroduced sika deer. The purpose of this study was to identify the genetic diversity and relationship of South Korean sika deer compared to those in other countries at Northeast Asia. Total 153 DNA samples (69 South Korean, 38 Chinese and 29 North Korean samples of captive bred, and 17 Russian of wild) were used in the analysis. The number of alleles per locus varied from 2~14 with a mean of 6. The expected heterozygosity and observed heterozygosity values were 0.3863 and 0.2740, respectively. The mean polymorphism information content value was 0.3528, ranging from 0.026~0.826. The genetic distance analysis showed genetic difference between South Korean and other populations, and Russian and North Korean populations were genetically similar. The maximum value of ΔK was observed when K was set to 2. When K=2, the genetic structure analysis revealed hybridization among four populations. Based on the information from structure results, we distinguished individuals with dominant genetic composition of a particular subspecies (>0.90), and the result showed relatively ‘pure’ individuals in South Korean population formed a distinct cluster that contradict other countries. As a result, all the sika deer populations in the four countries were genetically vulnerable. In addition, it is confirmed that relatively ‘pure’ individuals in South Korea were showed genetically different characteristics compared to other countries. However, the rate of hybridization was high in all populations of four countries, and it was also difficult to grasp the hybridization with other deer subspecies or other species. Moreover, STRUCTURE analysis results depending on the number and type of markers used for analysis. Therefore, additional samples and markers need to be acquired and analyzed.
L-carnitine supplementation improves the cryosurvival and subsequent development of bovine embryos
Kyeong Yeob Kim,Youn Bae Park,Byeong Ho Kim,Jin Hee Lee,Ji Hye Lee,Chanuka Kulatunga,Dong Eon Kim,Kyu Hyun Kim,Ryeong Eun Kim,Yoon Seok Nam,Min Kyu Kim 한국수정란이식학회 2018 한국수정란이식학회 학술대회 Vol.2018 No.11
Cryopreservation of bovine embryos is used to efficiently implant surrogate mothers. It has been widely accepted that high lipid content in the oocyte interrupts its survival during freeze-thaw cycles. Serum component in the culture medium is thought to increase the embryo`s lipid contents. Conversely, L-carnitine stimulates lipid metabolism by transporting long chain fatty acids into the mitochondria. Objective of this study was to analyze the effect of L-carnitine supplementation in IVM medium and defined IVC medium on the development, lipid contents and the cryosurvival of bovine IVF embryos. 0.0, 1.5, 3.0 and 6.0 mM L-carnitine was supplemented in IVM medium, respectively (IVM-LC 0.0, LC 1.5, LC 3.0 and LC 6.0). Development rate from the 2cell to the morula stages was higher in IVM-LC 3.0 groups than those of IVM-LC 6.0 (p<0.05). But there were no significant differences among the other groups in the blastocyst rates and lipid content results. When 0.0, 1.5, 3.0 and 6.0 mM L-carnitine were supplemented in IVC medium (IVC-LC 0.0, LC 1.5, LC 3.0 and LC 6.0), development competence was not significantly different between those embryos. Lipid contents of embryos treated L-carnitine (IVC-LC 1.5, 3.0 and 6.0) were significantly lower than embryos of non-treated group. L-carnitine was supplemented 0.0, 1.5, 3.0, 6.0 mM during IVM and 3.0 mM during IVC (LC 0.0 - 3.0, LC 1.5 – 3.0, LC 3.0 – 3.0, LC 6.0 – 3.0) and cryosurvival of blastocysts confirmed after freezing-thawing. There were no significant differences on development, but LC 3.0 – 3.0 was significantly lower lipid contents than other groups. And LC 3.0 – 3.0 had better survival rates and hatched rates of blastocysts than LC 0.0 – 0.0. In conclusion, supplementation of L-carnitine in defined IVC medium decreases lipid contents. And L-carnitine supplementation improves cryosurvival and developmental ability of bovine IVF embryos.