• The Korean Journal of Physiology and Pharmacology
• /
• v.24 no.1
• /
• pp.121-126
• /
• 2020

The ezrin-radixin-moesin (ERM) proteins are a family of membrane-associated proteins known to play roles in cell-shape determination as well as in signaling pathways. We have previously shown that amphetamine decreases phosphorylation levels of these proteins in the nucleus accumbens (NAcc), an important neuronal substrate mediating rewarding effects of drugs of abuse. In the present study, we further examined what molecular pathways may be involved in this process. By direct microinjection of LY294002, a PI3 kinase inhibitor, or of S9 peptide, a proposed GSK3β activator, into the NAcc core, we found that phosphorylation levels of ERM as well as of GSK3β in this site are simultaneously decreased. These results indicate that ERM proteins are under the regulation of Akt-GSK3β signaling pathway in the NAcc core. The present findings have a significant implication to a novel signal pathway possibly leading to structural plasticity in relation with drug addiction.

• Korean Journal of Animal Reproduction
• /
• v.19 no.2
• /
• pp.89-93
• /
• 1995

In this study was demonstrate that retrovirus-mediated gene transfer is one of the promising alternatives to the conventional pronuclear DNA microinjection approach, especially in transferring the exogenous genes into the boving embryos. By co-culturing of zona of zona-free one-cell stage embryos with the retrovirus-producing cells for 24 hours followed by 6 days of culture in virus-free medium, we could get morulae and blastocysts expressing the E. coli LacZ genes which were transferred by our retrovirus vector. The results obtained in this study are summarized as follows : 1. Addition of 5$\mu\textrm{g}$/ml of polybrene in the embryo and virus-producing cell co-culture medium did not affect development of zona-free one-cell embryo. 2. Compared with the intact embryos removal of zona at one-cell stage before co-culturing with the virus-producing cells for one day caused only slight decrease of embryo develpment. 3. Co-culture of 625 zona-free one-cell stage embryos with the virus-producing cells resulted in 65(10.4%) morulae or blastocysts, and 12.3%(8/65) of the morulae or blastocysts were E. coli LacZ positive.

• Molecules and Cells
• /
• v.25 no.1
• /
• pp.131-137
• /
• 2008

Crk-associated substrate (CAS) is a focal adhesion protein that is involved in integrin signaling and cell migration. CAS deficiency reduces the migration and spreading of cells, both of which are processes mediated by Rac activation. We examined the functions of v-Crk, the oncogene product of the CT10 virus p47gag-crk, which affects cell migration and spreading, membrane ruffling, and Rac activation in CAS-deficient mouse embryonic fibroblasts (CAS-/- MEFs). CAS-/- MEFs showed less spreading than did CAS+/+ MEFs, but spreading was recovered in mutant cells that expressed v-Crk (CAS-/-v-Crk MEF). We observed that the reduction in spreading was linked to the formation of membrane ruffles, which were accompanied by Rac activation. In CAS-/- MEFs, Rac activity was significantly reduced, and Rac was not localized to the membrane. In contrast, Rac was active and localized to the membrane in CAS-/-v-Crk MEFs. Lamellipodia protrusion and ruffle retraction velocities were both reduced in CAS-/- MEFs, but not in CAS-/-v-Crk MEFs. We also found that microinjection of anti-gag antibodies inhibited the migration of CAS-/-v-Crk MEFs. These findings indicate that v-Crk controls cell migration and membrane dynamics by activating Rac in CAS-deficient MEFs.

• Proceedings of the KSAR Conference
• /
• 2004.06a
• /
• pp.191-191
• /
• 2004

Transgenic animals are produced primarily by microinjecting exogenous DNA into the male pronuclei of a zygote. Microinjection method for gene transmitting is successful in mice but not efficient in farm animals, limiting it's general utility such as a large scale facility and labour. Based on our finding that sperm cells bind with exogenous DNA, sperm was used as a vector for producing transgenic animals to introduced green fluorescence protein(GFP) gene. (omitted)

• Korean Journal of Animal Reproduction
• /
• v.19 no.4
• /
• pp.293-305
• /
• 1996

Retrovirus는 DNA가 아닌 RNA를 유전 물질로 갖고 있는 동물 virus인데 각 virus는 RNA와 함께 크게 gag, pol. 그리고 env 등의 3가지 단백질로 구성되어 있다. gag 단백질은 virus의 내부구조를 형성하는 단백질이고, pol단백질은 감염을 통해 표적 세포에 도입된 retrovirus의 RNA를 DNA로 역전사시키는 reverse transcriptase의 역할을 하며, env단백질은 virusdml 외부를 구성하는 단백질로써 이 단백질에 의해 각 retrovirus의 종류에 따른 감염이 가능한 표적세포의 종류가 결정된다(host cell specificity). 따라서 어떤 retrovirus의 envelope단백질과 표식세포에 있는 retrovirus의 envelope 단백질에 대한 특정 receptor와의 상호 작용에 의해 세포속으로 도입된 virus의 RNA는 reverse transcriptase에 의해 DNA로 역전사된 후 표적세포의 genomic DNA에 삽입되는 특징을 가진다. 이러한 특징을 가진 retrovirus vector system은 형질 전환 동물의 생산에 있어서 현재까지의 주된 방법인 수정란의 pronucleus에의 DNA microinjection방법 보다 여러 가지 면에서 우수함에도 불구하고 쥐 이외의 다른 동물에서는 거의 이용되고 있지 않는 실정이다. 주된 원인으로는 현재 사용되고 있는 대부분의 retrovirus vector system이 쥐의 백혈병 virus를 근간으로 하기 때문에 이 system에서 생산된 virus는 쥐 이외의 다른 동물, 특히 유제류의 세포에는감염성이 아주 약하기 때문이다. 이러한 결점을 해결하기 위하여 최근에 기존의 쥐 백혈병 virus의 envelope protein을 vesicular stomatitis virus의 G protein으로 대체한 hybrid retrovirus vector system이 개발되었다. 이러한 system에서 생산되는 virus는 조류를 포함한 거의 모든 종류의 동물세포를 감염시킬 수 있으며 몇몇 특정세포에 대해서는 기존의 retrovirus vector system에 비해 1,000배 이상의 높은 감염도를 나타내는데 그 특징이 있다. 따라서 이러한 새로운 virus vector system을 이용할 경우, 보다 다양한 종에 있어서 형질전환 동물을 효율적으로 생산할 수 있을 뿐만 아니라 형질전환 동물의 생산 방법 자체를 다양화 시킬 수 있다고 본다.

• Korean Journal of Animal Reproduction
• /
• v.27 no.1
• /
• pp.1-7
• /
• 2003

Transgenic rabbits were produced by DNA microinjection using growth hormone receptor (GHR) and IGF-1 receptor (IGF-1R) genes fused to metallothionein(MT) promoter. The overall efficiencies for production of transgenic rabbits were 3.2% and 3.1% for GHR and IGF-lR genes, respectively. Founder rabbits transmitted transgenes to their progenies through medelian fashion. Growth rate of GHR and IGF-lR transgenic rabbits was significantly faster than that of non-transgenic rabbits. Transgenic rabbits grew large. (25% and 15% increase in body weight of GHR and IGF-lR transgenic rabbits, respectively) than non-transgenic rabbits and organ weight of transgenic rabbits increased, suggesting that GHR and IGF-1R genes affects growth rates in transgenic rabbits.

• Korean Journal of Animal Reproduction
• /
• v.19 no.3
• /
• pp.171-179
• /
• 1995

This study was carried out to investigate the fertilizing ability of round spematids isolated from seminiferous tubules. A round spermatid was introduced into the perivitelline space of a mature oocyte using Leitz micromanipulators and then subjected to electrofusion. Electrofusion was induced by applying a single DC pulse of 90V with a duration of 60$\mu$sec using Model 611 Square Wave Stimulator(Phipps and Bird, U.S.A) in 0.3 M sucrose fusion medium containing 0.05mM CaCl2 and 0.1mM MgSO4, Oocyte pre-activation was conducted by exposure to a single DC(80V, 80$\mu$sec) pulse in electrofusion medium at 1 hour before electrofusion. The incidence of fusion with pre-activated oocytes(23.8%, 57/239) was higher than that with nonactivated oocytes(6.7%, 3/45). The most of electro-stimulated mouse oocytes cleaved regardless of the success or failure of fusion. Karyotyping of embryos that developed into blastocysts after exposure to the fusion pulse were performe. We found that blastocysts from the fused oocytes were diploid whereas blastocysts from the unfused oocytes were haploid. About 11.7 and 11.5% of fused and unfused oocytes were developmental potentials of fused and unfused oocytes. Therefore, these results suggest that the mouse mture oocyte can be fertilized by fusion with a round spermtid and subsequently developed normally.

• Journal of Photoscience
• /
• v.1 no.1
• /
• pp.69-73
• /
• 1994

Sporangiophores (spphs) of Phycomyces blakesleeanus are positively phototropic to unilateral visible (blue) light over a range of fluence rates 10$^{-9}$ to 1 W/m$^2$. The maximal bending angle is always 70-75$\circ$ from the vertical. Many mutants with abnormal phototropism have been isolated. Complementation tests revealed that the genetic grouping is completely consistent with the phenotypic classification scheme, based on sensory responses other than those to light. The spph of the piloboloid mutant, the growth zone of which gradually ceases elongation but expands spherically, and the $\beta$-carotene-overproducing mutant show negative phototropism, in contrast to the wild type spph. We hypothesized that the phototropic orientation of spph is determined by the ratio of the maximal light fluenee rate at the proximal side to that at the distal side of the spph. Based on this hypothesis, we found that the maximal bending angle was larger in thin spphs than in thick ones, and larger in spphs containing smaller amount of $\beta$-carotene than in carotene-rich spphs. In addition to our hypothesis, gravitropic experiments revealed that the maximal bending angle of the wild type spph results from a balance among positive phototropism, negative gravitropism, and the optical properties of the spph. For further advancement of this study, we developed a mutant with a high proportion of uninucleate spores, and designed an efficient microinjection method for obtaining transformants.

• Natural Product Sciences
• /
• v.26 no.3
• /
• pp.221-229
• /
• 2020

This study was undertaken to investigate the protective effect of rice bran oil (RBO) on amyloid β protein (Aβ) (25-35)-induced memory impairment and brain damage in an ICR mouse model. Memory impairment was produced by intracerebroventricular microinjection of 15 nmol Aβ (25-35) and assessed using the passive avoidance test. Treatment with RBO at 0.1, 0.5, or 1 mL/kg (p.o. daily for 8 days) protected against Aβ (25-35)-induced memory impairment. Furthermore, Aβ (25-35)-induced decreases in glutathione and increases in lipid peroxidation and cholinesterase activity in brain tissue were inhibited by RBO, and Aβ (25-35)-induced increases of phosphorylated mitogen-activated protein kinases (MAPKs) and inflammatory factors, and changes in the levels of apoptosis-related proteins were significantly inhibited by RBO. Furthermore, Aβ (25-35) suppressed the PI3K/Akt pathway and the phosphorylation of CREB, but increased phosphorylation of tau (p-tau) in mice brain; these effects were significantly inhibited by administration of RBO. These results suggest that RBO inhibits Aβ (25-35)-induced memory impairment by inducing anti-apoptotic and anti-inflammatory effects, promoting PI3K/Akt/CREB signaling, and thus, inhibiting p-tau formation.

• Journal of Crop Science and Biotechnology
• /
• v.11 no.1
• /
• pp.13-16
• /
• 2008

Pepper fruit anthracnose caused by Colletotrichum species is an economically important disease that causes serious yield loss and quality deterioration in many Asian countries including South Korea and Taiwan. Recently, Capsicum baccatum PI594137 was found to exhibit broad-spectrum resistance to Colletotrichum acutatum. The inheritance of anthracnose resistance to C. acutatum was analyzed in an intraspecific population derived from a cross between C. baccatum Golden-aji and PI594137. Detached mature green fruits were inoculated using the microinjection method. The disease response was evaluated as the disease incidence and the overall lesion diameter at 7 days after inoculation(DAI). The segregation ratios of resistance and susceptibility to C. acutatum in the $F_2$ and $BC_s$ populations significantly fit a 3:1 Mendelian model. This result indicates that the resistance of PI594137 to C. acutatum is controlled by a single dominant gene.