• Reproductive and Developmental Biology
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• v.37 no.3
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• pp.149-154
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• 2013

The aim of this study was to evaluate the changes of protein patterns in granulosa cells and corpus luteum in ovaries during the estrus cycle in cows. The estrus cycle was devided into five steps of follicular, ovulatory, early-luteal, mid-luteal and late-luteal phases. In results, 61 spots of total 85 spots were repeated on follicular phase and 51 spots of total 114 spots were repeated on ovulatory phase. The 40 spots of total 129 spots were repeated on early-luteal phase and 49 spots of total 104 spots were repeated on mid-luteal phase. Also 41 spots of total 60 spots were repeated on late-luteal phase. On the other hands, the 16 spots were indicated difference in follicular phase and ovulation phase had a difference 10 spots. It was showed difference No. 103 spot in ovulation phase, No. 135 spot in early-luteal phase and No. 175 and 176 spots in mid-luteal phase. Also, the 11 spots were expressed specifically in mid-luteal phase and No. 178 and 179 spots were difference of expression in late-luteal phase. We confirmed that there were 7 spots for ovulation, 4 spots for luteinization and 2 spots for luteolysis. Spot No. 89~93 in ovulation phase were transferrin, and spot No.94~98 were HSP60. Spot No. 103 was Dusty PK, spot No. 135 was OGDC-E2, and spot No. 175 and 176 were Rab GDI beta from luteinization. Spot No. 178 and 179 in luteolysis were vimentin. This results suggest that will be help to basic data about infertility.

• Korean Journal of Animal Reproduction
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• v.25 no.1
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• pp.9-17
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• 2001

To determine the effect of the characteristics of the corpus luteum (CL) and serum metabolites on pregnancy rate following embryo transfer in Hanwoo cow, recipients were synchronized the estrus with CIDR. in vivo or in vitro produced bovine embryos were transferred to synchronized recipients. The characteristics of the CL were checked by rectal palpation and ultrasound. Blood samples were collected from jugular vein at Day 7 and analysed progesterone and serum metabolites. The results obtained from this study were as follows; 1. Total 397 embryos were transferred to recipient and 121 heads out of them were pregnant. The pregnancy rate was 30.5% and slightly differences between farms. 2. There was not significantly different the pregnancy rates between the size of CL and the cavity CL. In the recipients which had more than 2ng/$m\ell$ of serum progesterone level, the pregnancy rates was in higher than others (46.6% vs. 24.4%, 15.0%). 3. The highest pregnancy rate was obtained at 90 to 110mg/㎗ in serum total cholesterol, 14 to 16mg/㎗ in BUN, and 70 to 80mg/㎗ in serum glucose, respectively, and these metabolites in blood could be used as the criteria in the selection of recipient at embryo transfer.

• Journal of Embryo Transfer
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• v.17 no.1
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• pp.33-44
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• 2002

These studies were carried out to establish an effective in vivo embryo transfer methods in Hanwoo by analyzing several factors that influence this process. In an embryo transfer, recipients with grade A corpus luteum of the right ovary and that of grade B of the left one showed a higher pregnancy rate(p<0.1) than others. The pregnancy rates of frozen embryos were significantly lower(P<0.01) than those of fresh ones; the former resulting in 35% and the latter resulting in 56.2%. Transfer of embryos according to the estrus cycle(6.0 ∼ 9.0 days) did not show a significant difference in pregnancy rate with fresh embryos recording 45.4 ∼65.7% and frozen ones recording 22.0 ∼ 50.0%. According to the status of corpus luteum and embryo freezing or not, the pregnancy rate was higher on grade A corpus luteum with 40.8 ∼67.9% than B and C which ranged from 25.0∼56.0%. The results of embryo transfer according to the development stage and grade of embryos showed that regardless of the embryo's grade. transfer of morula recorded an average pregnancy rate of 46.3%. This results higher than the transfer of blastocyst which was 34.1%.

• Journal of Embryo Transfer
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• v.28 no.3
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• pp.169-175
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• 2013

Recently, the transgenic animal production technique is very important for the production of bio-parmaceutical as animal bio-reactor system. However, the absence of survival evaluation in vitro produced transgenic embryos has been a problem of the low productivity of transgenic animal because of absent of pre-estimate of pregnancy after transgenic embryos transferred into recipient. Therefore, this study is conducted to improve efficiency of transgenic cattle production by improving the non-surgical embryo transfer (ET) method. Transgenic bovine embryos were produced by injection of feline immunodeficiency virus enhanced green fluorescent protein (FIV-EGFP) lentiviral vector into perivitelline space of in vitro matured MII stage oocytes, and then in vitro fertilization (IVF) was occured. Normal IVF and EGFP expressing blastocysts were transferred into recipients. Results indicated that 2 expanded blastocysts (34.7%) transferred group showed significantly (P<0.05) higher pregnancy rate than 1 expanded blastocyst (26.8%) transferred group. In case of parity of recipient, ET to heifer (34.9%) showed significantly (P<0.05) higher pregnancy rate than ET to multiparous recipient (21.2%). However, there are no significant differences of pregnancy rate between natural induced estrus and artificial induced estrus groups. Significantly (P<0.05) higher pregnancy rate was obtained from recipient group which have normal corpus luteum with crown group (34.8%) than normal corpus luteum without crown (13.6%). Additionally, treatment of $100{\mu}g$ Gn-RH injection to recipient group (38.6%) 1 day before ET significantly (P<0.05) increase pregnancy rate than non- Gn-RH injection to recipient group (38.6%). We also transferred 2 EGFP expressing expanded blastocysts to each 19 recipients, 7 recipients were pregnant and finally 5 EGFP transgenic cattle were produced under described ET condition. Therefore, our result suggested that transfer of 2 good-quality expanded blastocysts to $100{\mu}g$ of Gn-RH injected recipient which have normal corpus luteum with crown is feasible to produce transgenic cattle.

• Korean Journal of Veterinary Service
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• v.27 no.1
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• pp.41-52
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• 2004

To establish the differential diagnosis and functional status in ovarian cystic cows, progesterone(P$_4$) and estrogen(E$_2$) level of cystic follicular fluid, ultrasonography for measuring the cystic diameter and thickness of cystic wall, and histological findings were investigated in cystic ovaries from slaughtered Korean native cows. Ovarian follicles were classified as systic if the diameter was greater than 25 mm by ultrasonography. Ovarian cysts < 3 mm of cystic wall thickness, < 10 ng/ml P$_4$ concentration and >10 ng/ml E$_2$ concentration were classified follicular cyst, ovarian cysts 3 mm of cystic wall thickness, 10 ng/ml P$_4$ concentration and <10 ng/ml E$_2$ concentration were classified luteal cyst, and ovarian cysts 3 mm of cystic wall thickness, < 10 ng/ml P$_4$ concentration and <10 ng/ml E$_2$ concentration were classified non-functional ovarian cyst, respectively. Also ovarian cysts were classified 8 types by anatomical and hisctological findings. Ovarian cysts with corpus luteum were 3 of 73 cows and ovarian cysts without corpus luteum were 70 cows. The incidence rates of 8 various types of ovarian cysts were as follows; 2Aa 56.2%, 2Ba 20.5% and 2Ab 15.1%, respectively. The incidence rates of ovarian cysts without corpus luteum were follicular cyst 76.7% and luteal cyst 19.2%. The thickness of cystic wall were lAb 3.9 mm, 2Ab 3.3 mm and 2Bb 3.2 mm, and the cystic fluid P$_4$ concentrations were above 10.0 ng/ml in lAb, 2Ab and 2Bb, respectively. There was significantly correlations between the thickness of cystic wall and cystic fluid P$_4$ concentration in ovarian cysts(p<0.05). The ovarian cyst was classified follicular cysts, luteal cyst and non-functional ovarian cyst by hormone analysis. The luteal cyst was accuratly dignosed by cystic wall thickness. But follicular cysts was misdiagnosed 13 cows of 56 cystic cows. The 13 cystic cows was determined as had non-fuctional ovarian cysts. The cystic fluid P$_4$ concentration was 3.3 ng/ml in follicular ovarian cysts and 30.1 ng/ml luteinized ovarian cysts. There was significantly positive correlations between thickness of cystic wall and serum P$_4$ concentration in follicular(r$^2$ =0.59, p<0.001) and luteal cysts(r$^2$=0.65, p<0.001). These results indicated that ovarian cysts had various stages of degeneration and luteal cyst was accuratly diagnosed measurement of cystic wall thickness by ultrasonography, but follicular cysts was not diagnosed only cystic diameter and cystic wall thickness. In conclusion, it is suggest that ovarian cysts was diagnosed by combination of clinical sign and anatomical cystic features.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.11
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• pp.1540-1545
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• 2012

Steroidogenesis requires coordination of the anabolic and catabolic pathways of lipid metabolism, but the profile of proteins associated with progesterone synthesis in cyclic and pregnant corpus luteum (CL) is not well-known in cattle. In Experiment 1, plasma progesterone level was monitored in cyclic cows (n = 5) and pregnant cows (n = 6; until d-90). A significant decline in the plasma progesterone level occurred at d-19 of cyclic cows. Progesterone level in abbatoir-derived luteal tissues was also determined at d 1 to 5, 6 to 13 and 14 to 20 of cyclic cows, and d-60 and -90 of pregnant cows (n = 5 each). Progesterone level in d-60 CL was not different from those in d 6 to 13 CL and d-90 CL, although the difference between d 6 to 13 and d-90 was significant. In Experiment 2, protein expression pattern in CL at d-90 (n = 4) was compared with that in CL of cyclic cows at d 6 to 13 (n = 5). Significant changes in the level of protein expression were detected in 32 protein spots by two-dimensional polyacrylamide gel electrophoresis (2-DE), and 23 of them were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Six proteins were found only in pregnant CL, while the other 17 proteins were found only in cyclic CL. Among the above 6 proteins, vimentin which is involved in the regulation of post-implantation development was included. Thus, the protein expression pattern in CL was disorientated from cyclic luteal phase to mid pregnancy, and alterations in specific CL protein expression may contribute to the maintenance of pregnancy in Korean native cows.

• Journal of Veterinary Clinics
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• v.15 no.2
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• pp.307-318
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• 1998

Accuracy of rectal palpation and ultrasonography for differential diagnosis of subestrous dairy trows were investigatedl using the result of pIRsma progesterone assay. The ovaries were examined 2 times of 10 days interval in 520 posearom and postinsemination subestroHs dairy cows, using rectal palpation and B-mode transrectal ultrasonography. The results of rectal palpation, ultrasonographic examination and measurement of plasma progesterone profiles in 520 subestrous dairy cows were silent brat or error of estrus detection 303 (58.3%), persistent corpus luteum 59 (11.3%), follicular cyst 37 (7.1%), luteal cyst 16 (3.1%), inactive ovary 9 (1.7%), granulosa tumor 1 (0.2%), hydmsalphinx 1 (0.2%), endomehris 81 (15.6%), pyometra 12 (2.3%) and mummified fetus 1 (0.2%), respectively. Accuncy of rectal palpation and ultrasonography for diagiosing ovarian disordeir based on plasma progesterone profiles were silent heat or error of estrus detection 80.5% and 96.7%$\boxUl$ persistent corpus luteum 57.6% and 94.9%, follicular cyst 62.5% and 91.9%1 luteal cyst 62.5% and 87.5%, maclive ovary 55.6% and 88.9% and granulosa cell tumor 100% and 100%, respectively. Acnuucy of rectal palpation for diagnosing uterine disorders based on ultrasonography was pyometra 75.0%1 endometritis 51.9% and mummified fetus 100%, respectively. Cbaracteristic ultrasonographic appearances of ovaries in subestrous dairy cows were as follows; Silent heat or error of estrus detection: anechoic follicle or hypoechoic corpus luteum than ovarian stroma was alternately present on Day 0 (first examination) and Day 10. Follicular cyst: uniformly nonechogenic ovarian structure $\geq $ 25 mm in diameter with a wall < 3 mm was present in ipsilateral on Day 0 and Day 10. Luteal cyst: luteal cyst was similar to follicular cysts but thickness of cystic wall was $\geq $ 3 mm. Inactive ovary : structures within ovaries was not present on Day 0 Bnd Day 10. Characteristic uthssonograpsc appearances of uterus in subestrous dairy cows were as follows; Endometritis: characterized by uterine lumen containing fluid in which 'snowy'echogenic particles art suspended. Pyometra: ultrasonographic appearance of pyometra was diffuse echogenic particles distributed in fluid within the distended uterus, and a thickened uterine wall. These results indicated that ultrasonography was practical far diagnosing reproductive disorders. To diagnosing ovarian disorders, ultrasonography should be carried out 2 times of 10 days interval and rndometritis should be differentiated with uterus of luteal phase in normal cycling cows.

• Journal of Embryo Transfer
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• v.27 no.3
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• pp.133-139
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• 2012

This study was carried out to develop the useful inducing method of estrus for Korean native cows. Under the condition of estrus induction by administering $PGF_{2{\alpha}}$ for the cows in which corpus luteum (CL) in ovaries was detected by ultrasonography, ovarian responses and the changes of progesterone ($P_4$) concentration against $PGF_{2{\alpha}}$ compared with conception rate were observed in cows and heifers. In inducing estrus administering $PGF_{2{\alpha}}$. to the cows which has corpus luteum in ovaries, ovarian reponses, the changes of progesterone concentration, and conception rate were identified and compared. The results attained from the studies were as follows. Significant decreases of CL in size over time after $PGF_{2{\alpha}}$ administration were detected in both cow and heifer groups (p<0.001), but not different between groups in the CL regression rate (p>0.05). In addition, the percentage changes relative to the plasma $P_4$ concentration on day 1 after $PGF_{2{\alpha}}$ treatment were decreased to below 1ng/ml. The growth rate of follicle was observed as 31% on day 1 and 42% on day 2 in cows, and 34% on day 1 and 97% on day 2 in heifers, resulting that growth of heifers are faster than that of cows (p<0.05). The conception rate after $PGF_{2{\alpha}}$ treatment were 60.5% and 64.2% in cows and heifers, respectively. It also indicated that the conception rate after estrus observation with $PGF_{2{\alpha}}$ injection was as high as 66.6% while that with timed-artificial insemination (TAI) regardless of the estrus observation was 56%, which means the pregnancy rate of artificial insemination after estrus observation was higher than that of TAI (p<0.05). In the result of all above, there were significant decreases in CL size and the plasma $P_4$ concentration by days but rapid growth in follicles, which has no differences in cows and heifers. The conception rate was commonly high after estrus observation and more than 50% under TAI.

• Korean Journal of Clinical Laboratory Science
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• v.50 no.4
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• pp.457-461
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• 2018

Corpus luteum (CL) is a transient endocrinal tissue that undergoes repeated formation and regression during the estrous cycle. In this study, we hypothesized that the functional and structural mechanism of angiogenesis is similar between CL and tumor formation. First, we measured mRNA and protein expression of angiogenic receptors (vascular endothelial growth factor receptor-2, VEGFR2; Tie 2) in the early, middle, and late phase CL tissues during the estrous cycle. Ral-interacting protein of 76 kDa (RLIP76) and H-ras mRNA and protein were also expressed in the CL tissues. VEGFR2 and Tie 2 mRNA and protein were expressed in the early and middle phase CLs and decreased in the late phase. H-ras mRNA and protein were expressed in the early and middle phase CLs, but not in the late phase. RLIP76 mRNA was expressed in all phase CLs, and the protein expression was highest in early phase CLs. We suggest that RLIP76 and H-ras, an oncogenic gene, regulate the function of the CL during the estrous cycle, and the proteins will play an important role in the angiogenic mechanism of the CL.

• Proceedings of the Korean Society of Food Hygiene and Safety Conference
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• 2001.10a
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• pp.204-207
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• 2001