• Title/Summary/Keyword: C3H mouse

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Molecular Cloning of H-Y Antigen Gene III. Construction of Mouse Testis cDNA Library and Screening of H-Y Ag Gene (H-Y 항원 유전자의 클로닝에 관한 연구 III. 생쥐정소 cDNA Library 구성과 유전자의 검색)

  • 이정렬;김창규;김종배
    • Korean Journal of Animal Reproduction
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    • v.17 no.1
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    • pp.43-48
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    • 1993
  • These experiments were carried out to construct mouse testis cDNA library and to to seen H-Y Ag gene. Mouse testis was obtained from BALB/c inbreed mouse that was after-born 1 week. Isolation of mouse testis total RNA was carried out by guanidum/cesium choloride, poly(A+) mRNAs were purified by oligo d(T)-cellulose chromatography method. To investigate protein synthesis activity, in-vitro translation carried out by total RNA and poly(A+) mRNA. The products of in-vitro translation were identified in 12.5% PAGE. Single strand DNA and double strand DNA were synthesized from poly(A+) mRNA and purified using phenol/chloroform/isoamylalcohol. Synthesized cDNA was combined with cohesive Eco RI polylinker, its recombination efficiencies were identified by X-gal and IPTG. In the cDNA library, 1$\times$107 phagemids were screened with 32P labelled probe. Hybridization were carried on $65^{\circ}C$ for 16~20hours. And 1$\times$106 phagemids were screened with rabbit-anti-H-Y. In former, select 5 positive clones, and later, 1 positive clone. Its southern blot analysis showed various size of insert cDNA from 0.7kb to 3kb.

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The Changes of Cyclic AMP Content by Opiates in Chronic Haloperidol Treated Mouse Striatum (Haloperidol 장기 투여된 Mouse Striatum에서 cAMP양에 미치는 Opiates의 영향)

  • Kim, Soo-Kyung
    • The Korean Journal of Pharmacology
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    • v.30 no.1
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    • pp.11-18
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    • 1994
  • Cyclic adenosine 3'5'-monophosphate (cyclic AMP) has been frequently accepted as an intracellular messenger for receptor-mediated action of opioids. In this experiment, it was designed to determine the interaction of dopaminergic and opioidergic system in the mouse striatum in normal and chronic haloperidol treated groups. Haloperidol 750ug/kg I.P. for 10 days was performed for dopamine denervation. The morphine, DAGO, DPDPE, and U5O,488H inhibited the increase of haloperidol-induced cyclic AMP content in chronic haloperidol treated mouse striatum. The inhibition of DAGO and DPDPE showed significant increase compared to normal mouse striatum. Naloxone showed antagonistic effect on the morphine and U5O,488H in chronic haloperidol treated group, and showed antagonistic effect on morphine, DAGO, DPDPE, and U5O, 488H in normal mouse striatum. These findings support that there is a functional interrelationship of dopaminergic and opioidergic pathway in the striatum. This result provides an evidence that following destruction of striatal dopaminergic neuron, there are some changes of cAMP content on the ${\mu},\;{\gamma},\;and\;{\kappa}$ opioid receptor, but the ${\kappa}$ opioid receptor still has its function.

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The Effects of T-2 Toxin on the Morphology of Mouse Organ (Mouse 장기의 형태에 미치는 T-2 Toxin의 영향)

  • 송재영;정덕화
    • Journal of Environmental Health Sciences
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    • v.21 no.4
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    • pp.96-101
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    • 1995
  • To study the immunotoxicity of mouse injected with fungal mycotoxin, T-2 toxin (Fusarium mycotoxin) was treated to 6 week-old female C3H/He mouse and the body, organ weight and morphological change were investigated. The weights of body, liver and kidney of mouse injected the 2 mg/kg of toxin was decreased to 17, 20 and 3%, respectively, compared to control animal and the comsumption of feed was also decreased with lapsing the time. The fat dropleting phenomenon and destruction of Golgi apparatus in liver and histopathological changes of tissue and mitochondria in small intestine were found by scanning electron microscopic observation.

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Overexpression of Mouse Nck Transforms Mouse Febroblast NIH3T3

  • Kim, Young H.;Han, Sun-Mi;Kim, Moon G.;Park, Dong-Eun;Park, Sang D.;Seong, Rho H.
    • Animal cells and systems
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    • v.1 no.3
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    • pp.521-526
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    • 1997
  • We isolated a mouse nck cDNA from the thymus cDNA expression library. The cDNA encodes a 377 amino acid protein and displays 97% amino acid sequence identity to human oncogenic protein nck, which is composed almost exclusivelv of three src homology 3 (SH3) domains and one SH2 domain. The sequence analysis also showed that the isolated cDNA is the mouse counterpart of the human nck and different from the mouse grb4, which has been reported to be highly similar to the human nck and, therefore considered as a mouse nck, Northern blot analysis showed that the transcript of the gene was 1.8 kb and was highly expressed in the testis, thymus, and brain but moderately in the liver and lymph node. Western blot analysis showed that the size of the protein was about 47 kDa. Overexpression of the mouse Nck transformed a mouse fibroblast cell line, NIH3T3. The results clearly indicate that normal nck gene has transforming ability and provide an argument against a suggested possibility that the transforming ability of the human nck gene is due to a mutation(s) in the gene.

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Properties and Activitiy Screening of Chromone Derivatives (크로몬 유도체의 물성 및 활성검색)

  • 김영로;이상현;김경순;정춘식;정재훈;김박광
    • YAKHAK HOEJI
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    • v.44 no.2
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    • pp.107-114
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    • 2000
  • We have synthesized 4-isonitroso-4H-1-benzopyran and 4-amino-2,3-dihydro -4H-1-benzopyran of chromone derivatives by using condensation method. Physico-chemical properties of these compounds were measured and analyzed by UV and HPLC method. The correlation coefficient of their methanol solutions by UV were 0.9992 and 0.9994, respectively. And oxime compound was resolved within 4 min and had a detection limit of 3 ng at S/N=3 by HPLC using a reversed phase column with three solvents(MeOH, $H_2O$, HAc). The amino compound was resolved within 4.5 min and had a detection limit of 10 ng at S/N=3 by HPLC under the same conditions. Anti-diabetic effect of chromone derivatives were investigated in the streptozotocin (STZ)-induced diabetic rats. Diabetes was induced in male Sprague-Dawley rats by injections of STZ (45 mg/kg, i.v). The investigation of the hair growth effect of isonitrosobenzopyran and 4-aminobenzopyran on the hair of black mouse (C57BL/6) was also carried out. The administraion of their ethanol solution to the black mouse (C57BU/6) through skin them promoted the growth of hair.

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Mucosal Mast Cell Responses in the Small Intestine of C3H/HeN and BALB/c Mice Infected with Echinostoma hortense

  • Ryang, Yong-Suk;Im, Jee-Aee;Kim, In-Sik;Kim, Keun-Ha
    • Biomedical Science Letters
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    • v.9 no.3
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    • pp.145-150
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    • 2003
  • In the intestinal mucosa, mast cells are thought to be responsible for the expulsion of parasites. We investigated the relationship of worm expulsion and mast cells in C3H/HeN and BALB/c mice infected with Echinostoma hortense. In addition, we examined whether the worm recovery rate was associated with the strain of mice, and whether a toluidine stain and immunohistochemistry using the c-kit antibody was effective in the detection of mast cells. In order to investigate the mucosal immune response of C3H/HeN and BALB/c mice, each mouse was infected orally with 30 E. hortense metacercariae. Then, the number of mucosal mast cells and worm recovery rates was observed in experimentally infected mouse strains between 1 week and 8 weeks post infection (PI). Mucosal mast cells were increased in 3 weeks P.I. in C3H/HeN and BALB/c mice. On the other hand, only mucosal goblet cells and worm recovery rates correlated in C3H/HeN mice (P=0.0482). Worm recoveries in C3H/HeN mice were 65.7$\pm$5.6, 53.3$\pm$5.4 and 6.7$\pm$0.6 in week 1, 2, and 3 P.I. and strongly decreased in week 3 P.I. Worm recoveries in BALB/c mice were 23.0$\pm$2.5, 10.0$\pm$1.0, and 6.7$\pm$0.6% in week 1, 2, and 3 P.I. and gradually decreased from week 1 P.I. to week 3 P.I. Worm recoveries in C3H/HeN mice were significantly higher than in BALB/c mice (P<0.00l). The number of mast cells in C3H/HeN and BALB/c mice using the anti-c-kit antibody reached to a peak in week 3 P.I. and recovered as normal level in week 5 P.I. and 6 P.I. The number in E. hortense-infected C3H/HeN mice (P=0.0015) was higher than in E. hortense-infected BALB/c mice (P=0.01) compared with the control group. There were significant differences in the number of mast cells among regions of the intestine in in C3H/HeN mice (P<0.05) but not in BALB/c mice (P>0.05). Immunohistochemistry using the anti-c-kit antibody was significant method as an examination of the number of mast cells (P=0.0002). In conclusion, the present study demonstrated that mast cells play an important role in worm recovery, and immunohistochemistry using the anti-c-kit antibody was superior to toluidine stain as an examination of mast cells.

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Anti-Angiogenic Activity of Mouse N-/C-terminal deleted Endostatin

  • Cho, Hee-Yeong;Kim, Woo-Jean;Lee, Sae-Won;Kim, Young-Mi;Choi, Eu-Yul;Park, Yong-Suk;Kwon, Young-Guen;Kim, Kyu-Won
    • BMB Reports
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    • v.34 no.3
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    • pp.206-211
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    • 2001
  • Endostatin, a proteolytic fragment of collagen XVIII, is a potent inhibitor of angiogenesis and the growth of several primary tumors. However, the opinions on the activity of endostatin derivatives deleted N- or C- terminal are still controversial. In this regard, we produced mouse endostatin and its derivatives in the prokaryotic system, and studied their anti-tumor activity. The [$^3H$]-thymidine incorporation assay demonstrated that N-terminal deleted mouse endostatin, and a C- and N-terminal deleted mutant, effectively inhibited the proliferation of human umbilical vein endothelial cells (HUVECs). The biological activity of endostatin was also shown by its in vivo anti-angiogenic ability on the chorioallantoic membrane (CAM) of a chick embryo. Treatment of $200\;{\mu}g$ of mouse endostatin, or N-terminal deleted mouse endostatin, inhibited capillary formation of CAM 45 to 71%, which is comparative to a 80% effect of positive control, $1\;{\mu}g$ of retinoic acid. An in vivo mouse tumor growth assay showed that N-terminal deleted mouse endostatin, and the N-/C-terminal deleted mutant, significantly repressed the growth of B16F10 melanoma cells in mice as did the full-length mouse endostatin. According to these results, N-and N-/C-terminal deleted mouse endostatins are the potent inhibitors of tumor growth and angiogenesis.

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The Effect of Cryopreservation on the Mouse Embryos at Various-pronuclear Stages

  • Park, M.C.;Kim, J.Y.;Kim, S.B.;Park, Y.S.;Park, H.D.;Lee, J.H.;Oh, D.S.;Kim, Jae-myeoung
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.2
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    • pp.174-180
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    • 2009
  • This study was carried out to establish an appropriate condition for the efficient cryopreservation of the mouse pronuclear embryo. In vitro cryopreservation of pronuclear embryos was carried out by slow freezing or vitrification methods and development rate of 2-cell, blastocyst and hatched blastocyst was measured as well as survival rate of the thawed pronuclear embryo. After slow freezing, vitrification and thawing of mouse pronuclear embryos, the survival rate and blastocyst development rate for the vitrification group was 97.3 and 53.4%, respectively, which was significantly higher as compared to the slow freezing group with 88.6 and 23.9%, respectively (p<0.05). Blastocyst developmental rate in each experimental group was significantly higher for 21 h in the post-hCG group at 40.5-57.0% than the 24 h post-hCG group at 40.5% (p<0.05). ICM (Inner cell mass) cell numbers of blastocyst-stage embryos during the different stages of mouse pronuclear embryos, slow freezing and vitrification period in the control and vitrification groups were 22.1${\pm}$2.7 and 17.0${\pm}$3.1-22.0${\pm}$3.2, respectively; hence, the slow freezing group (10.2${\pm}$2.0) had significantly higher cell numbers than those of the other two groups (p<0.05). Trophoblast (TE) cell number in the control group, 65.8${\pm}$12.6, was significantly higher than in the slow freezing group, 41.6${\pm}$11.1 (p<0.05). The total cell numbers in the control group and 21 h post hCG group were 87.9${\pm}$13.6 and 81.8${\pm}$14.1, respectively, and were significantly higher than for the slow freezing group (51.8${\pm}$12.6; p<0.05).

Prenatal Deaths and External Malformations Caused by X-Irradiation during the Preimplantation Period of ddy Mice (임신 ddy Mice에서 착상전기 방사선 조사에 따른 산전 사망 및 외부 기형 발현)

  • Ro, Hee-Jeong;Choi, Ihl-Bhong;Gu, Yeun-Wh
    • Radiation Oncology Journal
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    • v.16 no.3
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    • pp.233-243
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    • 1998
  • Purpose : To evaluate the effects of X-irradiation on prenatal deaths, i.e., preimplantation deaths, embryonic deaths and fetal deaths, and on external malformations in precompacted preimplantation ddy mice Materials and Methods : Pregnant mice (n=85) obtained by limiting the mating time to from 6 to 9 A.M., were segregated into 11 groups. The first five groups (n=26) were irradiated with X-ray doses of 0.1 0.5, 0.75, 1.5, and 3 Gy, respectively, at 24 h post conception (p.c.) of the preimplantation Period. The second five (n=27) groups were irradiated at the same X-ray doses, respectively, but at 48 h p.c. of the preimplantation period. The last group (n=32) was the control group. The uterine contents were examined on the 18th day of gestation for prenatal deaths and external malformations. Results : 1) A statistically significant increase in preimplantation deaths with increasing dose was observed in the experimental groups irradiated at 24 h p.c. and in the groups irradiated at 48 h p.c., as compared to the control group. The threshold dose was close to 0.05 Gy and 0.075 Gy for the irradiations at 24 h p.c. and 48 h p.c. respectively. 2) A statistically significant increase in embryonic deaths with increasing dose was observed in all irradiation groups, except the group irradiated with a dose of 0.1 Gy at 48 h p.c.. 3) No fetal deaths were found in any experimental group. 4) In the experimental groups irradiated at 24 h p.c. anomalies increased with statistical significance, as compared with the control group : 2 exencephalies, 2 open eyelids, 3 anophthalmias, 2 cleft Palates, 2 gastroschisis, 1 abdominal wall defect. 1 leg defect, and 2 short tail anomalies: the threshold dose for external malformations was close to 0.2 Gy at 24 h p.c.. In the groups irradiated at 48 h p.c., 1 open eyelid and 2 short tail anomalies were observed, but there was no statistical significance in those malformations. Conclusion : The results of this study reveal that X-irradiation of precompacted preimplantation ddy mice causes not only preimplantation deaths and embryonic deaths but also external malformations. In addition, external malformations were observed in our experiments at diagnostic doses, including 0.1 and 0.5 Gy. For this reason, we recommend that irradiation should be avoided during the preimplantation period by applying Rugh's 10-day rule.

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Cryopreservation of Mouse Embryos by Vitrification (Mouse 배의 Glass화 보존)

  • ;T.Kono;T. Nakahara
    • Korean Journal of Animal Reproduction
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    • v.13 no.2
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    • pp.63-69
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    • 1989
  • Eight-cell mouse embryos equilibrated with vitrification solution(VS3), consisting of glycerol and polyethlene glycol, were plunged directly into liquid nitrogen. The embryos were cryopreserved by vitrification without intra- and extracellular ice formation. After the vitrified embryos were warmed at 4$^{\circ}C$, sucrose dilution procedures were examined and the survival embryos were transferred to recipients after 48h incubation in vitro. The results were obtained as follows. 1. Mouse embryos equilibrated with VS3 were diluted with 1.5m sucrose-HB 1 solution, 0.5M sucrose-HB1 solution for 5min at room temperature, respectively. The proportion of vitrified-warmed embryos developed to blastocyst(85.7%) was as high as that of the embryos diluted with 1.04M sucrose-HB1 solution at 4$^{\circ}C$(85.5%). 2. Normal live youngs were obtained in 53.9%(55/102) of the vitrified-warmed embryos after tranfer to pseudopregnant recipients.

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