• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.60-60
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• 2003

소의 난포란과 난구세포의 체외배양시 plasminogen activators(PAs)의 생산을 보고하였다 (Choi 등, 1998). 따라서 본 연구는 돼지 난포란 및 난구세포의 체외성숙시 PAs의 생산을 SDS-PAGE와 Zymogram을 이용하여 조사하였다. PCOCs는 도축암퇘지의 난소로부터 채취하여, 난구세포가 충실한 것만 선별하였으며, 실험구에 사용될 난구세포는 pipetting에 의해 분리하여 이용하였다. 돼지 신선정액은 D-PBS로 1,500 rpm, 5분간 2회 원심분리하여 정장물질을 제거하고, 3회째는 5mM caffein이 함유된 BO(Brackett과 Oliphant, 1985) 배양액으로 세정하였다. 처리한 돼지 정액은 1$\times$$10^{8}$ cells/$m\ell$로 조정하여 20${\mu}\ell$씩 분주하고 0, 1, 2, 3 또는 4시간 동안 39$^{\circ}C$ 5% $CO_2$, 95% 공기인 배양기에서 수정능획득을 유도하였다. 배양이 완료된 정액은 20${\mu}\ell$의 sample buffer(5% SDS, 20% glycerol, 0.0025% bromophenol blue 그리고 0.125M Tris HC1 buffer)에 넣어 -7$0^{\circ}C$ 동결기에 보관하였다. 전기영동은 4% stacking gel과 10% separating gel로 분리하였으며, 20 mA에서 90분간 실시하였다. Zymogram은 Choi 등(1988)의 방법에 따라 실시하여 PAs의 생산을 확인하였으며, 이상의 실험은 3반복을 실시하였다. 시험구 전체에서 urokinase type plasminogen activator(uPA)가 확인되었으며, 체외수정능 획득시간에는 차이가 없었다. 두 종류의 고분자량의 uPA의존성 영역이 나타났으며, 분자량은 65kD과 62 kD이었다. 이러한 결과로 볼 때 Hart 등(1986)이 uPA의 경우 다양한 영역의 분자량 변이를 확인할 수 있었다고 한 것과 동일하였으며, 돼지 정자가 체외수정능 획득시 uPA를 생산하는 것을 확인할 수 있었다.

• Journal of Embryo Transfer
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• v.31 no.1
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• pp.53-59
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• 2016

This study was to investigate effect of progesterone ($P_4$) on prostaglandin (PG) synthases and plasminogen activators (PAs) system in bovine endometrium during estrous cycle. Endometrium tissues were collected from bovine uterus on follicular and luteal phase and were incubated with culture medium containing 0 (Control), 0.2, 2, 20 and 200 ng/ml $P_4$ for 24 h. The $PGF_{2{\alpha}}$ synthase (PGFS), $PGE_2$ synthase (PGES), cyclooxygenase-2 (COX-2), urokinase PA (uPA), and PA inhibitors 1 (PAI-1) mRNA in bovine endometrium were analyzed using reverse transcription PCR and PA activity was measured using spectrophotometry. In results, COX-2 was higher at 2 ng/ml $P_4$ group than control group in luteal phase (p<0.05), but, it did not change in follicular phase. Contrastively, PGES was significantly increased in 2 ng/ml $P_4$ group compared to control group in follicular phase, but there were no significant differ among the treatments in luteal phase. uPA was no significant difference between $P_4$ treatment groups and control group in both of different phase. PAI-1 was decreased in 20 ng/ml $P_4$ group compared to control group in follicular phase (p<0.05). PA activity was decreased in 2 ng/ml $P_4$ group compared to other groups in follicular and luteal phase (p<0.05). In conclusion, we suggest that $P_4$ may influence to translation and post-translation process of PG production and PA activation in bovine endometrium.

• Journal of Embryo Transfer
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• v.14 no.1
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• pp.16-19
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• 1999

• 한국생물공학회:학술대회논문집
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• 2005.10a
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• pp.604-605
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• 2005

• Proceedings of the Korean Society of Radiological Science
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• 1995.10a
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• pp.148-149
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• 1995

• Journal of Life Science
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• v.20 no.6
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• pp.838-844
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• 2010

A fibrinolytic enzyme of Streptomyces corcohrussi from soil sediment was purified by chromatography using DEAE-Sephadex A-50 and Sephadex G-50. The analysis of SDS-polyacrylamide gel suggested that the purified enzyme is a homogeneous protein and the molecular mass is approximately 34 kDa. The purified enzyme showed activity of 0.8 U/ml in a plasminogen-rich fibrin plate, while its activity in a plasminogen-free fibrin plate was only 0.36 U/ml. These results suggested that the purified enzyme acts as a plasminogen activator. The fibrinolytic activity of the enzyme under the supplementation of protease inhibitors, $\varepsilon$-ACA, t-AMCHA and mercuric chloride in the enzyme reaction was less than 24%, indicating that it could be modulated by the plasmin and/or fibrinogen inhibitors involved in the fibrinogen-to-fibrin converting process. As time passed, $Zn^{2+}$, a heavy metal ion, inhibited the activity to 34.1%. The optimum temperature of the purified enzyme was approximately $50^{\circ}C$ and over 92% of the enzyme activity was maintained between pH 5.0 and 8.0. Therefore, our results provide a potential fibrinolytic enzyme as a noble thrombolytic agent from S. corcohrussi.

• Reproductive and Developmental Biology
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• v.35 no.4
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• pp.463-468
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• 2011

This study investigated the changes of plasminogen activators (PAs) activity, expression and localization of tissue plasminogen activator (tPA) and urokinase plasminogen activator (uPA) during the estrous cycle in pigs. Estrous cycle was sorted into three group by pre-ovulation (Pre-Ov), post-ovulation (Post-Ov) and early to mid-luteal stages (Early to mid-L). Analysis for immunohistochemistry was confirmed by location of tPA and uPA. Porcine uterus tissue was cut into $1{\times}1$ cm squares, and were incubated in DMEM/F-12 medium for 1 h at $38^{\circ}C$, 5% $CO_2$ for measurement of PA activity. Western blotting was implemented for measurement of PA quantity. In results, the blood vessels and secretory glands were increased in Post-Ov stage than Pre-Ov and Early to mid-L stages. The tPA and uPA was located mainly within lumen of blood vessels and secretory glands. The PA activity in Post-Ov ($0.99{\pm}0.03$) stage were significantly (p<0.01) higher than Pre-Ov stage ($0.51{\pm}0.03$) and Early to mid-L stage ($0.21{\pm}0.04$). Expression of PAs were significantly (p<0.05) higher in Early to mid-L stage than other stages. These results indicate that PAs activity and expression may change in uterus tissue during the estrous cycle in pigs.

• Korean Journal of Veterinary Research
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• v.24 no.2
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• pp.137-147
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• 1984

Cells with an epithelioid morphology were isolated from the rabbit myometrium and were grown in culture. The cells had a doubling time of 53 hours when grown in the presence of 10% fetal calf serum in Basal Eagle's medium with 3mM glutamine. In the presence of estrogen plus insulin, doubling time was reduced to 40 hours. Creatine kinase activity upon reaching confluency was determined to be 0.019 unit per mg protein. Approximately 30% of the activity was extractable only in high ionic strength buffer. Cells also contained plasminogen activator with a specific activity of 140 CTA units per million cells. Creatine kinase was mainly BB form. The cells contained a cross reactive protein against bovine smooth muscle uterine anti-myosin.

• Molecular & Cellular Toxicology
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• v.2 no.3
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• pp.212-215
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• 2006

Several reports have suggested a possible relationship between blood coagulation factors and schizophrenia. Plasminogen activator inhibitor type 1 (PAI-1) belongs to a serine protease inhibitor family, which regulates fibrinolysis and proteolysis by inhibiting plasminogen activation. The purpose of this study was to investigate the association of polymorphisms of the PAI-1 gene with schizophrenia in Korean population. Two important polymorphisms (-675 4G/5G and -844 G/A) located on promoter region of the PAI-1 gene were analyzed on 178 schizophrenia patients and 226 controls. The genotypic and allelic associations of -675 4G/5G were found significant. Furthermore, haplotype analysis revealed significant result, which suggests that -675 4G/5G polymorphism might confer increased susceptibility for schizophrenia in Korean population.

• Journal of Life Science
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• v.15 no.3 s.70
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• pp.434-438
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• 2005

In the previous study, we isolated a myulchikinase (MK), which has fibrinolytic activity and cytotoxicity to the tumor cell line, from myl- chi-jeot-gal. In this study, the effect of NaCl concentration, metallic ions, pH, temperature, and plasminogen on the activity of MK was analysed. The MK activity was maintained at least $80\%$ activity up to $30\%$ NaCl, which indicates that the enzyme may be halotolerant. The optimal pH and temperature were 8 and $40^{\circ}C$, respectively. The fibrinolytic activity of MK was completely inhibited with 0.5 mM $Hg^{2+}$ and inhibited to $50^{\circ}C$ with 1 mM $Cu^{2+}\;and\;Zn^{2+}$. The MK showed strong activity in plasminogen- rich fibrin plate but not in plasminogen-free fibrin plate. The result indicates that the MK may be a plasminogen activator type fibrinolytic enzyme.