• Title/Summary/Keyword: Microinjection

Search Result 202, Processing Time 0.053 seconds

성장관련 유전자를 이용한 형질전화토끼의 생산

  • 진동일
    • Proceedings of the Korean Society of Embryo Transfer Conference
    • /
    • 2000.11a
    • /
    • pp.46-54
    • /
    • 2000
  • Transgenic rabbits were produced by DNA microinjection using growth hormone receptor (GHR) and IGF-1 receptor (IGF-1R) genes. Overall efficiencies for production of transgenic rabbits were 3.2% and 3.1% in GHR and IGF-1R genes, respectively. Founder rabbits transmitted transgenes to their progenies through medelian fashion. Growth rate in GHR and ICF-1R transgenic rabbits was faster than non-transgenic rabbits. Transgenic rabbits grew larger (25% and 15% increase in body weight of GHR and IGF-1R transgenic rabbits, respectively) than non-transgenic rabbits and organ weight of transgenic rabbits increased, suggesting that GHR and IGF-1 genes affects growth rates in transgenic rabbits.

  • PDF

Time-dependent Modulation of Cerebral Ischemic Injury by Activated Macrophages/microglia after Lipopolysaccharide Microinjection into Rat Corpus Callosum

  • Lee, Jae-Chul;Kim, Won-Ki
    • Proceedings of the PSK Conference
    • /
    • 2003.10a
    • /
    • pp.106-107
    • /
    • 2003
  • tIn brain ischemic insult, inflammatory cells such as macrophages and lymphocytes are chemo-attracted into the brain lesion and release cytokines, resulting in an activation of microglia that are functionally equivalent to peripheral macrophages in the central nervous system. In cerebral ischemic insults, activated inflammatory cells such as microglia and macrophages may be implicated in the pattern and degree of ischemic injury by producing various bioactive mediators. (omitted)

  • PDF

Electroacupuncture Analgesia Is Improved by Adenoviral Gene Transfer of Dopamine Beta-hydroxylase into the Hypothalamus of Rats

  • Kim, Soo-Jeong;Chung, Eun Sook;Lee, Jun-Ho;Lee, Chang Hoon;Kim, Sun Kwang;Lee, Hye-Jung;Bae, Hyunsu
    • The Korean Journal of Physiology and Pharmacology
    • /
    • v.17 no.6
    • /
    • pp.505-510
    • /
    • 2013
  • Electroacupuncture (EA) is a modified form of acupuncture that utilizes electrical stimulation. We previously showed that EA stimulated rats were divided into responders that were sensitive to EA and non-responders that were insensitive to EA based on the tail flick latency (TFL) test. The dopamine beta-hydroxylase (DBH) gene was more abundantly expressed in the hypothalamus of responder rats than non-responder rats. To determine whether overexpression of DBH gene expression in the hypothalamus modulate EA analgesia, we constructed a DBH encoding adenovirus and which was then injected into the hypothalamus of SD rats. Microinjection of DBH or control GFP virus into the hypothalamus had no changes on the basal pain threshold measured by a TFL test without EA treatment. However, the analgesic effect of EA was significantly enhanced from seven days after microinjection of the DBH virus, but not after injection of the control GFP virus. DBH expression was significantly higher in the hypothalamus of DBH virus injected rat than control GFP virus or PBS injected rats. Moreover, expression of the DBH gene did not affect the body core temperature, body weight, motor function or learning and memory ability. Although the functional role of DBH in the hypothalamus in the analgesic effect of EA remains unclear, our findings suggest that expression of the DBH gene in the hypothalamus promotes EA analgesia without obvious side-effects.

Fabrication and Modeling of Microlens Array by a Modified LIGA Process

  • Kim Dong Sung;Lee Hyun Sup;Yang Sang Sik;Lee Bong-Kee;Lee Sung-Keun;Kwon Tai Hun;Lee Seung S.
    • Proceedings of the Korean Society for Technology of Plasticity Conference
    • /
    • 2003.04a
    • /
    • pp.7-13
    • /
    • 2003
  • Microlens arrays were fabricated using a novel fabrication technology based on the exposure of a PMMA (Polymethylmethacrylate) sheet to deep X-rays and subsequent thermal treatment. X-ray irradiation causes the decrease of molecular weight of PMMA, which in turn decreases the glass transition temperature and consequently causes a net volume increase during the thermal cycle resulting in a swollen microlens. A new physical modeling and analyses for micro lens formation were presented according to experimental procedure. A simple analysis based on the new model is found to be capable of predicting the shapes of micro lens which depend on the thermal treatment. For the replication of micro lens arrays having various diameters with different foci on the same surface, the hot embossing and the microinjection molding processes has been successfully utilized with a mold insert that is fabricated by Ni-electroplating based on a PMMA microstructure of micro lenses. Fabricated microlenses showed good surface roughness with the order of 1nm.

  • PDF

Microinjection of Glutamate into the Amygdala Modulates Nociceptive and Cardiovascular Response in Freely Moving Rats

  • Ahn, Dong-Kuk;Kim, Yun-Sook;Park, Jae-Sik
    • The Korean Journal of Physiology and Pharmacology
    • /
    • v.2 no.6
    • /
    • pp.687-693
    • /
    • 1998
  • This study was performed to examine the mean arterial pressure and nociceptive jaw opening reflex after microinjection of glutamate into the amygdala in freely moving rats, and to investigate the mechanisms of antinociceptive action of amygdala. Animals were anesthetized with pentobarbital sodium (40 mg/kg, ip). A stainless steel guide cannula (26 gauge) was implanted in the amygdala and lateral ventricle. Stimulating and recording electrodes were implanted into each of the incisor pulp and anterior digastric muscle. Electrodes were led subcutaneously to the miniature cranial connector sealed on the top of the skull with acrylic resin. After 48 hours of recovery from surgery, mean arterial pressure and digastric electromyogram (dEMG) were monitored in freely moving rats. Electrical shocks (200 ${\mu}sec$ duration, $0.5{\sim}2$ mA intensity) were delivered at 0.5 Hz to the dental pulp every 2 minutes. After injection of 0.35 M glutamate into the amygdala, mean arterial pressure was increased by $8{\pm}2$ mmHg and dEMG was suppressed to $71{\pm}5%$ of the control. Injection of 0.7 M glutamate elevated mean arterial pressure by $25{\pm}5$ mmHg and suppressed dEMG to $20{\pm}7%$ of the control. The suppression of dEMG were maintained for 30 minutes. Naloxone, an opioid receptor antagonist, inhibited the suppression of dEMG elicited by amygdaloid injection of glutamate from $28{\pm}4\;to\;68{\pm}5%$ of the control. Methysergide, a serotonin receptor antagonist, also inhibited the suppression of dEMG from $33{\pm}5\;to\;79{\pm}4%$ of the control. However, phentolamine, an ${\alpha}-adrenergic$ receptor antagonist, did not affect the suppression of dEMG. These results suggest that the amygdala can modulate both cardiovascular and nociceptive responses and that the antinociception of amygdala seems to be attributed to an augmentation of descending inhibitory influences on nociceptive pathways via serotonergic and opioid pathways.

  • PDF

Comparison of In Vitro Development of Porcine Embryos Derived from Transfer of Embryonic Germ Cell Nuclei into Oocytes by Electrofusion and Piezo-Driven Microinjection

  • Ahn, Kwang-Sung;Won, Ji-Young;Heo, Soon-Young;Kang, Jee-Hyun;Shim, Ho-Sup
    • Reproductive and Developmental Biology
    • /
    • v.31 no.2
    • /
    • pp.127-131
    • /
    • 2007
  • Embryonic germ (EG) cells are undifferentiated stern cells isolated from cultured primordial germ cells (PGC). These cells share many characteristics with embryonic stem cells including morphology and pluripotency. Undifferentiated porcine EG cell lines demonstrating capacities of differentiation both in vitro and in vivo have been established. Since EG cells can be cultured indefinitely in an undifferentiated state, whereas somatic cells in primary culture are often unstable and have limited lifespan, EG cells may provide inexhaustible source of karyoplasts in nuclear transfer (NT). In this study the efficiencies of NT using porcine EG and fetal fibroblast cells were compared. Two different techniques were used to perform NT. With conventional NT procedure (Roslin method) involving fusion of donor cells with enucleated oocytes, the rates of development to the blastocyst stage in EG and somatic cell NT were 16.8% (59/351) and 14.5% (98/677), respectively. In piezo-driven microinjection (Honolulu method) of donor nuclei into enucleated oocytes, the rates of blastocyst formation in EG and somatic cell NT were 11.9% (15/126) and 9.4% (9/96), respectively. Regardless of NT methods used in this study, EG cell NT gave rise to comparable rate of blastocyst development to somatic cell NT. Overall, EG cells can be used as karyoplast donor in NT procedure, and embryos can be produced by EG cell NT that may be used as an alternative to conventional somatic cell NT.

Approaches to Improving Production Efficiencies of Transgenic Animals

  • Tojo, Hideaki
    • Proceedings of the KSAR Conference
    • /
    • 2000.10a
    • /
    • pp.7-8
    • /
    • 2000
  • Transgenic animals are very useful for scientific, pharmaceutical, and agricultural purposes. In livestock, transgenic technology has been used forthe genetic alteration of farm animals, the production of human proteins inlarge quantities in the milk of transgenic farm animals, and the generation of animals with organs suitable for xenotransplantation. To date, the transfer of foreign genes into farm animals has been performed mainly by microinjection of DNA into the pronuclei of fertilized eggs. However, the overall success rate of transgenic animals in livestock so far has been disappointingly low, eg., the efficiency is 0∼5% in swine, and less than 1% in sheep and cattle, compared with the rate in mice where 5% microinjected develop into transgenic animals. Recently, McGreath et al. (2000) have succeeded in producing the gene targeted sheep by the use of nuclear transfer from cultured somatic cells transfected with a foreign gene in vitro. However, we may need plenty of time until currently employ this method for gene transfer to farm animals. We have been studying to exploit the method for improving production efficiencies of transgenic animals with emphasis of its application to farm animals. The present paper describes three approaches that we have made in our laboratory to improve production efficiencies of transgenic animals, based on the DNA microinjection method. 1. Co-injection of restriction enzyme with foreign DNA into the pronucleus for elevating production efficiencies of transgenic animals. 2. Efficient selection of transgenic mouse embryos using EGFP as a marker gene. 3. Phenotypes of tansgenic mice expressing WAP/hGH-CAG/EGFP fusion gene produced by selecting transgenic embryos. 4. Efficient site-specific integration of the transgene targeting an endogenous lox like site in early mouse embryos.

  • PDF

Study on Production of Transgenic Pig Harboring Tissue Plasminogen Activator Gene

  • Park, Jin-Ki;Jeon, Ik-Soo;Lee, Yun-Keun;Lee, Poongyeon;Kim, Sung-Woo;Kim, Jung-Ho;Han, Joo-Hee;Park, Chun-Gyu;Min, Kwan-Sik
    • Proceedings of the KSAR Conference
    • /
    • 2003.06a
    • /
    • pp.43-43
    • /
    • 2003
  • This study was conducted to produce transgenic pig harboring human tissue plasminogene activator (tPA) gene. Two different tPA genes containing bovine $\beta$-casein promoter and mouse uroplakin promoter were prepared for microinjection and confirmed the expression level of tPA protein from the CHO (Chinese hamster ovary) cell lines by gene transfection. Concentration of tPA expression from the six cell lines (all of CHO cells) were average 212.4 ng/ml. Reconstructed DNA to used the CHO cell were microinjected into the pronuclei of in vivo embryos The total of 2,307 zygotes were collected from 95 donors and 1,851 embryos were in 1-cell stage which were visualized the pronuclei for DNA microinjection. The concentration of linear DNA was 2.0 ng per microliter and injected into zygotes with two pronuclei on an inverted Nikon microscope equipped with narishige micromanipulator and modulation contrast optics. The 541 embryos injected with bovine $\beta$-casein promoter-tPA were transferred to 22 recipients. The 1,154 embryos injected with mouse uroplakin promoter-tPA were transferred to 51 recipients. Sixty nine offspring from 9 delivered sows were produced. We analysed the transgenes with PCR methods from 69 offsprings, but could not detect the PCR product from piglet tails DNA.

  • PDF