• Proceedings of the KSAR Conference
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• 2004.06a
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• pp.224-224
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• 2004

Plasminogen activators (PAs)는 자궁분비액, 난포액, 정장물질 등을 포함한 여러 가지 세포외 분비액(extracellular fluids) 및 plasma에 풍부하게 존재하는 세포외 전구효소인 plasminogen을 plasmin으로 전환시키는 단백질분해효소이다. PAs는 섬유소용해, 배란, 착상 및 수정을 포함한 다양한 생리적인 과정에서 중요한 역할을 수행하는 것으로 알려져 있다. 따라서, 본 연구는 돼지의 체외수정 과정시 정자와 난자에서의 plasminogen activators activity의 변화를 SDS-PAGE와 zymography를 이용하여 검토하였다. (중략)

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2006.10a
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• pp.38-39
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• 2006

• Reproductive and Developmental Biology
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• v.29 no.2
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• pp.75-82
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• 2005

This study were examined whether plasminogen activators (PAs) are produced by porcine fresh or frozen-thawed cumulus-oocytes complexes (COCs) and cumulus cell free-oocytes. In fresh or frozen-thawed COCs and oocytes for 0 hour cultured, no activity of PAs was detected. However, at 24 hours of culture urokinase-type plasminogen activator (uPA) was detected in COCs and denuded oocytes. In the frozen-thawed COCs and cumulus cell free-oocytes cultured for 24 hours, no PAs were observed. After COCs were cultured for 48 hours, tissue-type plasminogen activator (tPA) and tPA-PAI were observed in COCs only. In the frozen-thawed COCs and cumulus cell free-oocytes cultured for 48 hours, no PAs were observed. These results suggest that uPA, tPA and tPA-PAI are produced by porcine COCs, but only uPA by oocytes during maturation for 24 hours. Only tPA, and tPA-PAI are produced by COCs cultured for 48 hours, and no PAs are produced by denuded-oocytes cultured for 48 hours. In all of the frozen-thawed groups, no PAs are observed by COCs and denuded-oocytes.

• Reproductive and Developmental Biology
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• v.38 no.2
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• pp.63-70
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• 2014

Plasminogen activators (PAs) are serine proteases that convert plasminogen to plasmin. Two type of PAs are urokinase-type PA (uPA) and tissue-type PA (tPA). Plasminogen is present in most extracellular fluids. PAs play in various reproductive processes including implantation, ovulation and fertilization. In the spermatozoa, PAs and PAIs play a role in sperm motility and fertilization. PAs in the sertoli cell are stimulated spermatozoa maturation and sperm activation through the phospholipase A2. The oocyte maturation is the process for fertilization and implantation. PAs in cumulus-oocyte complexes (COCs) are related to oocyte maturation by protein kinase A and C. In the ovulatory process, PAs activity are changed and it are related to reducing the tensile strength of ovarian follicle wall. The uterine environment is important for reproduction and the uterus undergo tissue remodeling. In the uterus and oviduct of mammals, expression and activity of PAs are changed during estrous cycle. Thus, expression and activity of PAs are concerned to many reproductive functions. Therefore, PAs seem to important factor of regulator in reproductive events.

• Reproductive and Developmental Biology
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• v.32 no.2
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• pp.97-104
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• 2008

Plasminogen activators (PAs) are serine protease that cleave plasminogen to form the active protease plasmin. PA/plasmin system playa role in mammalian fertilization and motility and acrosome reaction of sperm. The present study was undertaken to identify PAs in porcine gametes and investigate a possible role of plasminogen in in vitro fertilization in the pig. When boar spermatozoa were preincubated in a fertilization medium (mTBM) for 0, 2, 4 or 6 h, the activity of tPA-PAI ($110{\sim}117\;kDa$), tPA ($62{\sim}70\;kDa$), and uPA ($34{\sim}38\;kDa$) was observed in the sperm incubation medium and sperm sample. PA activities in the sperm incubation medium significantly (p<0.05) increased according to increasing incubation times, while PA activities in sperm significantly (p<0.05) decreased at the same times. In addition, the rate of acrosome reaction in spermatozoa increased by increasing culture times. When oocytes were separated from porcine cumulus-oocytes complexes at 0, 22 or 44 h of maturation culture, no PA activities were observed in cumulus free-oocyte just after aspiration from follicles. However, the activity of tPA-PAI ($108{\sim}113\;kDa$) and tPA ($75{\sim}83\;kDa$) was observed at 22 h of in vitro culture and significantly (p<0.05) increased as the duration of the culture increased. On the other hand, when porcine oocytes were activated by sperm penetration or calcium ionophore, plasminogen significantly (p<0.05) increased ZP dissolution time (sec) in activated oocytes by sperm penetration. These results suggest that supplementation of plasminogen to fertilization medium may playa positive role in the improvement of in vitro fertilization ability in the pig.

• Proceedings of the KSAR Conference
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• 2004.06a
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• pp.251-251
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• 2004

Plasminogen activators (PA)는 다수의 세포 형태에서 분비되는 것으로 알려진 serine protease이다. PA는 섬유소 용해, 배란, 유선 퇴화, 착상 및 수정 등 다양한 생리적인 과정에 관여한다. 본 연구는 난포액이 돼지 난자의 체외성숙에 미치는 영향을 검토하기 위하여, 다양한 조건하에서의 돼지 난자의 성숙과 난구세포-난자 복합체(Cumulus-Oocyte complexes: COCs) 또는 conditioned medium 내의 PA 활성을 검토하였다. 직경 2∼6m 난포로부터 COCs를 회수하여 일부는 난구세포를 제거하였다. (중략)

• Reproductive and Developmental Biology
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• v.32 no.2
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• pp.89-95
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• 2008

The present study was undertaken to identify changes of plasminogen activators (PAs) in porcine oviductal epithelial cells (POECs) during the estrous cycle classified with post-ovulatory stages (Post-Ov), early to mid-luteal stages (Early-mid L) and pre-ovulatory (Pre-Ov) stages. The urokinase-type plasminogen activator (uPA) was only observed on day 5 and day 7 of culture in the POECs on all the estrous cycles and gradually increased according to increasing culture times, but not Early-mid L. In POECs-conditioned medium, uPA, tissue-type (tPA) and tPA-PA inhibitor (tPA-PAI) activity were observed at all culture times during estrous cycles. The uPA activity of POECs-conditioned medium on Post-Ov stage were significantly (p<0.05) decreased during prolonged cultures. On the other hand, the tPA activity of POECs-conditioned medium at Post-Ov stage was significantly (p<0.05) higher on day 5 than compared to the other days. Although was higher on day 1 at Post-Ov stage, the tPA-PAI activity of POECs-conditioned medium was significantly (p<0.05) higher on day 7 at all stage than that of day 5 of the culture. Taken together, these results suggest that uPA, tPA and tPA-PAI are produced by POECs, and the variations of the PAs activity are regulated in the different stages of the estrous cycle.

• Journal of Embryo Transfer
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• v.28 no.3
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• pp.273-280
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• 2013

The aim of this study was to establish a three dimensional (3D) culture system of endometrial cells and to examine the plasminogen activators (PAs) activity in porcine uterine. The 3D culture system in porcine endometrial cells was composed to mixture 3D gel, stromal cells and epithelial cells. The 3D culture system was used to identify normal structure search as uterine tissue and PAs expression in this study. In results, porcine endometrium epithelial cells forming a top monolayer and endometrium stromal cells developed as fibroblast-like within 3D matrix scaffold. Expression of urokinase-type PA (uPA) and tissue-type PA (tPA) were observed during the 3D culture using immunofluorescence. PA activity in 3D-cultured endometrial cells was no significant difference between the tissue type, but 2D culture system were significantly lower than in 3D-cultured endometrial cells (P<0.05). Therefore, basic system and functional aspect of 3D culture could be established with similar system of endometrium tissue. We suggest that this study was assumed applicable as baseline data to investigate mechanism between porcine uterus cells in vitro.

• Journal of Embryo Transfer
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• v.28 no.3
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• pp.257-263
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• 2013

The endometrium undergoes a cyclic growth and tissue remodeling as changes of epithelial cells, and plasminogen activators (PAs) are related to endometrium tissue remodeling. This study was to evulate expression of urokinase-type plasminogen activator (uPA) and tissue-type plasminogen activator (tPA) in porcine uterine epithelial cells. In results, the uPA and tPA were expressed in uterine tissue, epithelium and secretory glands in porcine endometrial cell. In addition, the uPA and tPA were expressed in cultured epithelial cells, and it were mainly expressed in cytoplasm. In porcine uterine tissue and epithelial cells, uPA activity was higher than activity in tPA. In PAs mRNA expression levels, uPA mRNA level was significantly higher than tPA mRNA level (P<0.05). The fluorescence intensity of uPA protein was also higher than fluorescence intensity of tPA protein, and uPA protein expression was significantly higher than in tPA protein expression (P<0.05). Therefore, we suggest that a physiological function in porcine uterine epithelial cells should be more influenced by uPA than in tPA during pre-ovulatory phase.

• 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.