• Title/Summary/Keyword: Transgenic animal

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Effects of fermented ginseng on memory impairment and β-amyloid reduction in Alzheimer's disease experimental models

  • Kim, Joonki;Kim, Sung Hun;Lee, Deuk-Sik;Lee, Dong-Jin;Kim, Soo-Hyun;Chung, Sungkwon;Yang, Hyun Ok
    • Journal of Ginseng Research
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    • v.37 no.1
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    • pp.100-107
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    • 2013
  • This study examined the effect of fermented ginseng (FG) on memory impairment and ${\beta}$-amyloid ($A{\beta}$) reduction in models of Alzheimer's disease (AD) in vitro and in vivo. FG extract was prepared by steaming and fermenting ginseng. In vitro assessment measured soluble $A{\beta}42$ levels in HeLa cells, which stably express the Swedish mutant form of amyloid precursor protein. After 8 h incubation with the FG extract, the level of soluble $A{\beta}42$ was reduced. For behavioral assessments, the passive avoidance test was used for the scopolamine-injected ICR mouse model, and the Morris water maze was used for a transgenic (TG) mouse model, which exhibits impaired memory function and increased $A{\beta}42$ level in the brain. FG extract was treated for 2 wk or 4 mo on ICR and TG mice, respectively. FG extract treatment resulted in a significant recovery of memory function in both animal models. Brain soluble $A{\beta}42$ levels measured from the cerebral cortex of TG mice were significantly reduced by the FG extract treatment. These findings suggest that FG extract can protect the brain from increased levels of $A{\beta}42$ protein, which results in enhanced behavioral memory function, thus, suggesting that FG extract may be an effective preventive or treatment for AD.

Intestinal Strangulation Associated with Pubic Fracture in a Dog (개에서 치골골절과 연관된 장폐색)

  • Lee, Jae-Il;Kim, Young-Suk;Jeong, Seong-Mok;Kim, Myung-Jin;Park, Chang-Sik;Kim, Myung-Cheol
    • Journal of Veterinary Clinics
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    • v.23 no.4
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    • pp.458-460
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    • 2006
  • A 2-year-old female, 3.8kg weight, mixed-breed dog was presented for a 10-day history of vomiting and anorexia. The dog was diagnosed with pelvic fracture by vehicular accident at a local animal hospital before 2 months, but fracture did not reconstructed. After 2 months, the dog suffered from intermittent vomiting and anorexia. On barium contrast, we diagnosed the dog as intestinal obstruction. Consequently, we were found obstruction by strangulation and adhesion of intestine at the fracture site through the operation. At present after 18 months of surgery, general physical condition of the dog is good.

Comparative study of wound healing in porcine urinary bladder with CO2 laser and scalpel incisions (CO2 Laser와 Scalpel을 이용한 절개 시 돼지 방광에서의 창상치유 평가)

  • Lee, Jae-Yeon;Jeong, Seong-Mok;Cho, Sung-Whan;Park, Chang-Sik;Kim, Myung-Cheol
    • Korean Journal of Veterinary Research
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    • v.50 no.3
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    • pp.247-251
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    • 2010
  • This study compared the instrument performance and tissue healing of a steel scalpel with a $CO_2$ laser in an animal urinary bladder surgery model. Landrace and Yorkshire mixed breed pigs were used. Two symmetrical incisions were made in urinary bladder of each pig. One incision was made on the left side of ventral aspect on urinary bladder using a steel scalpel, while the other incision was performed on the right side using a $CO_2$ laser with an 8W output power. Each instrument was evaluated clinically for speed, ease of incision, and extent of bleeding. At 7 and 21 days after initial wounding, each wound was taken for histological observations. The scalpel was an easier instrument to use in the confines of the urinary bladder tissue, compared with the laser. However, there was no significant difference between the two groups. The amount of bleeding was less in the laser group but the time of the incisions was shorter with the scalpel. Scalpel incisions showed complete restoration of the epithelium and muscularis. On the other hand, the laser incisions showed incomplete restoration of the epithelium and muscularis. However, most of wound healing in the laser incisions was accomplished according to the time lapse. Although the scalpel produced less damage to the urinary bladder tissue and was easier to handle than the $CO_2$ laser, it did not provide hemostasis that was helpful for use on highly vascular tissue. The $CO_2$ laser provided good hemostasis, but delayed wound healing. In conclusion, the $CO_2$ laser provided better hemostasis and better surgical field than the scalpel. The $CO_2$ laser was used effectively in urinary bladder incision.

Migration Activity of Chicken Gonadal Primordial Germ Cells (gPGCs) and Post-transfer Localization of LacZ-transfected gPGCs in the Embryonic Gonads

  • Jeong, D.K.;Han, J.Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.9
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    • pp.1227-1231
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    • 2002
  • A powerful tool for chicken transgenesis could be established by employing a germline chimera production through primordial germ cell transplantation. This study was conducted to examine whether foreign gene-transfected gonadal primordial germ cells (gPGCs) have a migration activity into the gonad after transfer to recipient embryos. In Experiment 1, gPGCs of Korean Ogol Chicken were retrieved from 5.5-day-old embryos and subsequently transferred to the dorsal aorta of 2.5-day-old White Leghorn embryos after being labeled with PKH26 fluorescent dye. To confirm migration activity after transplantation, recipient embryos were sacrificed and examined on 3 days after transfer. Sex determination was concomitantly undertaken to examine whether sex of recipient embryos could affect the migration activity of gPGCs. All of embryonic gonads examined showed positive signals with PKH26 fluorescence and W-chromosome specific band by polymerase chain reaction (PCR) was detected in male embryos when gPGCs with ZW chromosome were transferred to recipient embryos. In Experiment 2, retrieved gPGCs were transfected with LacZ gene-containing cytomegalovirus promoter ($pCMV{\beta}$) by electroporation and subsequently transferred to recipient embryos. LacZ gene expression was identified in the gonads of 6 or 10-day-old recipient embryos and hatched-chicks. A total of 20 embryos and 12 hatched-chicks were examined and 11 of them (10 embryos and one hatched chicken; 11/32=34.4%) expressed $\beta$-galactosidase, a marker substance of LacZ gene. The results of this study demonstrated that foreign gene-transfected gPGCs can migrate and settle down into the gonad after being transferred into the blood vessel of the recipient embryos. This established technique will contribute to developing a peer biotechnology for transgenic chicken.

Establishment and characterization of porcine mammary gland epithelial cell line using three dimensional culture system (3차원 배양 시스템을 이용한 돼지 유선 상피 세포 주 특성과 설정)

  • Chung, Hak-Jae
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.18 no.10
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    • pp.551-558
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    • 2017
  • To study and validate tissue-specific promoters and vectors, it is important to develop cell culture systems that retain the tissue and species specificity. Such systems are attractive alternatives to transgenic animal models. This study established a line of porcine mammary gland epithelial cells (PMECs) from a primary culture based on the cellular morphology and mRNA levels of porcine beta-casein (CSN2). The selected PMECs were stained with the cytokeratin antibody, and were shown to express milk protein genes (CSN2, lactoferrin, and whey acidic protein). In addition, to confirm the acini structure of PMEC932-7 in 3D culture, live cells were stained with SYTO-13 dye, which binds to nucleic acid. The acini of these PMECs on matrigel were formed by the aggregation of peripheral cells and featured a hollow lumens. The system was demonstrated by testing the effects of the culture conditions to cell culture including cell density and matrigel methods of the PMECs. These results suggest that PMECs possess the genetic and structural features of mammary epithelial cells.

Development of Transgenic Fish for the Production of Human EGF Protein (내재적 유전자에 의한 어류난자에서의 hEGE 단백질 생산을 위한 기술개발)

  • 황창남;송기철;이재현;윤종만;김기동;이상호;박홍양
    • Korean Journal of Animal Reproduction
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    • v.25 no.3
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    • pp.277-286
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    • 2001
  • Improvement and possible commercialization of a home-made electroporation apparatus(home-made) were further tried to establish a simple and effective introduction of foreign gene into sperm followed by in vitro fertilization. Expressions of introduced pJJ9 and pNT plasmids were shown in all fertilized eggs with electroporated spermatozoa. In particular, with this gene transfer system all the fry showed a consistently transient expression in the syncytium of the yolk sac. This fact is important since some required, minute quantity of human proteins can be produced from the established transient expression on the yolk sac of all fry derived from in vitro fertilization with electroporated spermatozoa. To explore tissue-specific expression in fish, which we will use a similar system later, we targeted the nerve tissue to see whether tissue-specific promoter is working in fish properly. pNT plasmid containing a nerve cell-specific tubulin promoter gene demonstrated consistently exact targeted expressions among the developing nerve cells in later stages of embryos and hatched fry. Finally, liver-specific genes are now being cloned by using already selected primers for useful human protein gene fusion.

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Acupuncture analgesia for operation in Korean native goats

  • Lee, Soo-jin;Park, Chang-sik;Jun, Moo-hyung;Kim, Nam-joong;Lee, Jae-il;Kim, Young-suk;Kim, Myung-jin;Lee, Jae-yon;Jeong, Seong-mok;Kim, Duck-hwan;Kim, Myung-cheol
    • Korean Journal of Veterinary Research
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    • v.45 no.4
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    • pp.581-585
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    • 2005
  • The purpose of this study is to establish the analgesic effects of electroacupuncture for Korean native goat. Electroacupuncture was applied to the 6 Korean native goats. In 3 of them, rumenotomy was performed, and in the other 3, laparotomy was done. The analgesic induction time was 15 to 30 minutes. The acupoints used were Tian-ping (Celestial Peace, GV-5), Bai-hui (Hundred Meetings, GV-20), left 13th thoracic nerve and left 3rd lumbar nerve. Electroacupuncture was performed in lateral recumbency. Needles were inserted 1-2 cm deep, and connected to the electroacupuncture apparatus. The electrical stimulation condition was 30 Hz and 2-6 volts. Initially, the voltage of analgesia mode was maximized in each channel. And, the output was slowly reduced to the critical point that goats could tolerate without obvious discomfort or pain. Surgical operation was done successfully under electroacupuncture analgesia in 6 Korean native goats. In addition, the changes of temperature, heart rate and respiratory rate were studied during acupuncture analgesia. For 3 months after surgery, no experimental animals showed clinical problem in 6 Korean native goats.

Gene Transfer into Chicken Embryos using Defective Retroviral Vectors Packaged with Vesicular Stomatitis Virus G Glycoprotein Envelopes (Vesicular Stomatitis Virus G Glycoprotein Envelope으로 포장된 Defective Retroviral Vector를 이용한 닭의 배로의 유전자 전이)

  • 권모선;임은정;허영태;이훈택;이영만;김태완
    • Korean Journal of Animal Reproduction
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    • v.25 no.2
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    • pp.171-180
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    • 2001
  • Compared to other gene transfer system, the advantages of retrovirus-mediated gene transfer are technical ease, efficient expression and genetic stability. Despite the high potency of the retrovirus vector system in gene transfer, one of the drawbacks is a difficulty in concentration of virus stock. To overcome this problem, we tested a new retrovirus vector system producing the progeny retrovirus particles encapsidated with VSV-G (vesicular stomatitis virus G glycoprotein). The infectivity of this virus was not sacrificed by ultracentrifugal concentration and the host cell range extended from all mammalian to fish embryos. Virus titer after 1,000 x concentration was more than 10$^{8}$ CFU/ $m\ell$ on most of the target cell lines. We applied this pantropic viruses in transgenic chicken production by injecting the concentrated (100$\times$) stock into subgerminal cavity of stage X chicken embryos. The survival rate of chicken embryos after injection was about 20% and gene integration rate in surviving embryos was scored almost 100%. Analyses of RT-PCR and fluorescence microscopy, however, showed no evidence of the transgene expression.

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Impaired Hippocampal Synaptic Plasticity and Enhanced Excitatory Transmission in a Novel Animal Model of Autism Spectrum Disorders with Telomerase Reverse Transcriptase Overexpression

  • Rhee, Jeehae;Park, Kwanghoon;Kim, Ki Chan;Shin, Chan Young;Chung, ChiHye
    • Molecules and Cells
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    • v.41 no.5
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    • pp.486-494
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    • 2018
  • Recently, we have reported that animals with telomerase reverse transcriptase (TERT) overexpression exhibit reduced social interaction, decreased preference for novel social interaction and poor nest-building behaviors-symptoms that mirror those observed in human autism spectrum disorders (ASD). Overexpression of TERT also alters the excitatory/inhibitory (E/I) ratio in the medial prefrontal cortex. However, the effects of TERT overexpression on hippocampal-dependent learning and synaptic efficacy have not been investigated. In the present study, we employed electrophysiological approaches in combination with behavioral analysis to examine hippocampal function of TERT transgenic (TERT-tg) mice and FVB controls. We found that TERT overexpression results in enhanced hippocampal excitation with no changes in inhibition and significantly impairs long-term synaptic plasticity. Interestingly, the expression levels of phosphorylated CREB and phosphorylated $CaMKII{\alpha}$ were significantly decreased while the expression level of $CaMKII{\alpha}$ was slightly increased in the hippocampus of TERT-overexpressing mice. Our observations highlight the importance of TERT in normal synaptic function and behavior and provide additional information on a novel animal model of ASD associated with TERT overexpression.

Factors Affecting Primary Culture of Nuclear Transfer Blastocysts for Isolation of Embryonic Stem Cells in Miniature Pigs

  • Kim, Min-Jeong;Ahn, Kwang-Sung;Kim, Young-June;Shim, Ho-Sup
    • Reproductive and Developmental Biology
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    • v.33 no.3
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    • pp.133-137
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    • 2009
  • Pluripotent embryonic stem (ES) cells isolated from inner cell mass (ICM) of blastocyst-stage embryos are capable of differentiating into various cell lineages and demonstrate germ-line transmission in experimentally produced chimeras. These cells have a great potential as tools for transgenic animal production, screening of newly-developed drugs, and cell therapy. Miniature pigs, selectively bred pigs for small size, offer several advantages over large breed pigs in biomedical research including human disease model and xenotransplantation. In the present study, factors affecting primary culture of somatic cell nuclear transfer blastocysts from miniature pigs for isolation of ES cells were investigated. Formation of primary colonies occurred only on STO cells in human ES medium. In contrast, no ICM outgrowth was observed on mouse embryonic fibroblasts (MEF) in porcine ES medium. Plating intact blastocysts and isolated ICM resulted in comparable attachment on feeder layer and primary colony formation. After subculture of ES-like colonies, two putative ES cell lines were isolated. Colonies of putative ES cells morphologically resembled murine ES cells. These cells were maintained in culture up to three passages, but lost by spontaneous differentiation. The present study demonstrates factors involved in the early stage of nuclear transfer ES cell isolation in miniature pigs. However, long-term maintenance and characterization of nuclear transfer ES cells in miniature pigs are remained to be done in further studies.