• Asian-Australasian Journal of Animal Sciences
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• 제30권8호
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• pp.1086-1092
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• 2017

Objective: O-linked N-acetylglucosamine (O-GlcNAc) transferase (OGT) catalyzes the addition of O-GlcNAc and GlcNAcylation has extensive crosstalk with phosphorylation to regulate signaling and transcription. Pig OGT is located near the region of chromosome X that affects follicle stimulating hormone level and testes size. The objective of this study was to find the variations of OGT between European and Chinese pigs. Methods: Pigs were tested initially for polymorphism in OGT among European and Chinese pigs by polymerase chain reaction and sequencing at the U.S. Meat Animal Research Center (USMARC). The polymorphism was also determined in an independent population of pigs including European and Chinese Meishan (ME) breeds at the National Institute of Animal Science (NIAS, RDA, Korea). Results: The intron 20 of OGT from European and Chinese pigs was 514 and 233 bp, respectively, in the pigs tested initially. They included 1 White composite (WC) boar and 7 sows ($2Minzu{\times}WC$, $2Duroc\;[DU]{\times}WC$, $2ME{\times}WC$, $1Fengzing{\times}WC$) at USMARC. The 281-bp difference was due to an inserted 276-bp element and GACTT in European pigs. When additional WC and ME boars, the grandparents that were used to generate the $1/2ME{\times}1/2WC$ parents, and the 84 boars of 16 litters from mating of $1/2ME{\times}1/2WC$ parents were analyzed, the breeds of origin of X chromosome quantitative trait locus (QTL) were confirmed. The polymorphism was determined in an independent population of pigs including DU, Landrace, Yorkshire, and ME breeds at NIAS. OGT was placed at position 67 cM on the chromosome X of the USMARC swine linkage map. Conclusion: There was complete concordance with the insertion in European pigs at USMARC and NIAS. This polymorphism could be a useful marker to identify the breed of origin of X chromosome QTL in pigs produced by crossbreeding Chinese and European pigs.

• Journal of the Korean Academy of Clinical Electrophysiology
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• 제6권1호
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• pp.81-89
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• 2008

Purpose: This study aims to examine effect of microcurrent electrostimulation on burn healing by electric intensity and of which the electric intensity on the acute burn being cured with microcurrent electrostimulation therapy. Methods: 28 Sprague Dawley Rats is classified into a control group of 8 rats, an experimental group I of 10 rats and an experimental group II of 10 rats. The control group is not cured, the experimental group I is exposed to 10 Hz, and $100{\mu}A$ with microcurrent electrostimulation, and the experimental group II is exposed to 10 Hz, $300{\mu}A$ for 15 minutes a day. The next day, 2th, 4th, and 6th day after rats is burned. Result: There are not significant differences of length change of the burn cure between the control group, the experimental group I, and the experimental group II by a period. However, systematically hair follicle cell on the 2th day and epidermal cell on the 6th day turn up in the experimental group I, and the experimental group II. Inquiry: Nancy(1994) did not obtain the desired result when the skin of a pig is exposed to 0.1 Hz, and $100{\mu}A$ for wound healing. In the result of the study, when burn length is measured on the 2th, 4th, and 6th to see the length change of acute burn, there is not significant differences among 3 groups. Conclusion: Statistically, there is not significant differences of the length change between 3 groups. However, systematically the burn is cured faster in the experimental group I, and the experimental group II than in the contrast group.

• Journal of Embryo Transfer
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• 제11권3호
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• pp.233-240
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• 1996

This study was carried out to determine the effect of cumulus cell attachment and various factors on in vitro maturation of pig foflicular oocytes. Oocytes with various configuration of cumulus cell mass were collected ftom ovaries of mature gilts by asperating with syringe equipped with needles of different gauges, follicle size and with or without cumulus cells. They were cultured in TCM-199 mediun containing FGS(fetal calf serum) for 30~48 hours in incubator with air containing 5% $CO_2$ at 38.5$^{\circ}C$. Mter orcein staining at in vitro maturation condition, GV, GVBD, anaphase, telophase and M II were observed. Results are surumarized as follows: 1. Recovery rates were 55.8, 55.5 and 34.4% when the cumulus-compacted oocytes were collected with 18, 21, 26 gauge needles of syringes, respectively. 2. 79% of oocytes with compacted cumulus cells were at GV stage and most of the oocytes with partially denuded and denuded cumulus cells were from GVBD to M- II stages. 3. Percentage of mature oocytes among those which are follicular diameter of 1~2, 3~6 and over 6 mm was 42.6, 53.2 and 60.8%, respectively. 4. Percentage of mature oocytes among those which are compacted, partially denuded and denuded was 60.5, 46.2 and 35.4% respectively. 5. Percentage of mature oocytes in co-cultured with monolayers of cumulus cells was higher (57.1%) than that found with oocytes cultured alone (53.4%).

• Journal of Ginseng Research
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• 제34권4호
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• pp.327-335
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• 2010

We reported previously that the administration of Korean red ginseng water extract (KRG-WE) protected the guinea pig testis against damage induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin (a potent endocrine disruptor). We also found that crude saponin from ginseng was the active ingredient responsible for this protection. Here, we examined the biological role of KRG-WE in an animal model of age-induced dysfunction of spermatogenesis. Twenty-four male Sprague-Dawley (six 2-month-old and eighteen 12-month-old) rats were used. The young and old control groups received only vehicle. The ginseng saponin (GS)- and KRG-WE-treated groups received GS (40 mg/kg body weight/day) and KRG-WE (200 mg/kg body weight/day), respectively, for 4 months. The number of cells, Sertoli cell index, Johnsen's score, and sex hormone levels decreased significantly with age. However, the administration of KRG-WE and GS markedly improved the number of germ cells, seminiferous tubular size, and Johnsen's score in the old rats. Ginseng produced a distinct testicular histological improvement in old rats. KRG-WE and GS elevated testosterone levels, while attenuating the aberrant increase in follicle stimulating hormone and luteinizing hormone levels. Sperm kinematics evaluated by a computer-assisted sperm analyzer demonstrated improvement in the percentage of motile sperm, progressive sperm motility, and curvilinear velocity associated with sperm quality, supporting the beneficial role of red ginseng in senile spermatogenesis. Overall, the total water extract had a more potent effect than the corresponding saponin fraction. In conclusion, Korean red ginseng rejuvenated age-induced testicular dysfunction. Additionally, the total water extract was more potent than the corresponding saponin fraction.

• Clinical and Experimental Reproductive Medicine
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• 제32권3호
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• pp.199-206
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• 2005

연구목적: 난포가 폐쇄되는 동안의 생화학적 변화를 규명하기 위하여 미성숙 돼지의 정상 및 폐쇄 난포 내 lactate dehydrogenase (LDH) 활성도 변화 및 동일 난포액 내 스테로이드호르몬의 농도변화를 알아보기 위하여 본 연구를 시행하였다. 재료 및 방법: 난포액 (FF), 과립세포 (GC), 협막세포 (TC) 내 LDH의 활성도를 측정하였으며, 난포액 내 progesterone ($P_4$), testosterone (T), estradiol ($E_2$)의 농도변화를 방사면역측정법으로 정량하였다. 결　과: 정상 및 폐쇄 난포에서 $P_4$의 농도변화를 보이지 않았다. 그러나 폐쇄 난포액 내 T의 농도($3.85{\pm}1.50ng/ml$)는 정상 난포 ($1.29{\pm}0.54ng/ml$)에 비해 현저히 높았으며 정상 난포 내 $E_2$의 농도 ($43.29{\pm}19.51ng/ml$) 는 폐쇄 난포 ($18.82{\pm}7.27ng/ml$)에 비해 현저히 높은 것으로 나타났다. 정상 난포액 내 $P_4$의 농도는 난포의 크기에 정의 상관관계 (r=0.75)를 보였다. 정상 난포 내 T:$P_4$의 비율 ($8.14{\pm}3.35$)은 폐쇄 난포 ($1.39{\pm}0.60$)에 비해 현저히 높았으며, 정상 TC ($433.63{\pm}102.40{\mu}U/{\mu}g$ DNA) 및 FF ($246.86{\pm}58.96{\mu}U/{\mu}l$) 내 LDH 활성도는 폐쇄 난포 (각각 $83.7{\pm}10.5$와 $38.71{\pm}9.00$)에 비해 현저히 높게 나타났다. 정상 난포의 GC 및 FF 내 LDH 활성도는 $E_2$의 농도와 부의 상관관계를 보였지만, 폐쇄 난포의 TC 내 LDH 활성도는 $P_4$, T, $E_2$의 농도에 대해 정의 상관관계를 나타내었다. 결　론: 본 실험의 결과, 미성숙 돼지 난포의 폐쇄는 TC 내 LDH 활성도 감소와 밀접한 관계를 갖는 것으로 사료된다.

• Journal of Animal Science and Technology
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• 제46권4호
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• pp.571-584
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• 2004

It is known widely that granulosa cell apoptosis leads follicular atresia and macrophages exert their effects directly and/or indirectly from the initiation to the completion of follicular atresia by phagocytosis of apoptotic bodies and secretion of various cytokines. However, the site of initiation, propagation routes and the elimination methods of apoptotic bodies, and the time and methods of penetration of macrophages into the follicles are not known completely. Using pig(Yorkshire-breed) ovary, immunohistochemical studies with TUNEL for apoptotic bodies and pig macrophage monoclonal antibody 4E9 for macrophages, and light and transmission electron microscopic observations were performed. In the pig, follicular atresia began with the granulosa cell apoptosis, and the apoptosis of theca intema cells occured at the same time. The apoptosis of granulosa cells initiated randomly within the granulosa cell layer and propagated rapidly into the whole layer. Ultrastructura1ly, apoptotic granulosa cells showed characteristic pyknotic and deformed nucleus and intracytoplasmic vesicles. Apoptotic bodies were eliminated by intact granulosa cells and macrophages. Intact granulosa cells ingested apoptotic bodies transiently, soon after they fell into the apoptosis. Finally, apoptotic bodies and degenerated oocyte were phagocytosed by macrophages. Macrophages entered the ovarian follicle at the time of initiation of granulosa cell apoptosis, and migrated with the progression of apoptosis. By elimination of theca cells, macrophages contributed the completion of follicular atresia These results will provide valuable informations on the study of the interrelation between macrophage and ovarian follicular atresia.

• Asian-Australasian Journal of Animal Sciences
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• 제17권3호
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• pp.317-324
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• 2004

The present study was conducted to investigate the involvement of nitric oxide (NO) in cumulus expansion, oocyte mortality and meiotic maturation of porcine cumulus enclosed oocytes (CEOs) cultured in two different models when gonadotropins, including follicle-stimulating hormone (FSH) and human chorionic gonadotropin (hCG) were presented or not. And the interaction between NO and $\beta$-mercaptoethanol ($\beta$-ME), a free radical scavenger was also investigated. Two models refer to spontaneous maturation model and hypoxanthine (HX) medium model. All the 3,433 eligible CEOs were incubated at $39^{\circ}C$ and the cumulus expansion, oocyte morphology and nuclear phase were evaluated 44 h after incubation. (1) In spontaneous maturation model, NO stimulates the cumulus expansion and $\beta$-ME delayed it. NO doesn't affect the oocyte meiotic resumption but inhibits the oocytes to develop to metaphase II. (2) In HX medium model, NO or $\beta$-ME doesn't affect the expansion in the absence of gonadotropins, but in the presence of gonadotropins, NO or $\beta$-ME inhibits the expansion. In the presence of gonadotropins, NO inhibits the oocyte meiotic resumption and it especially inhibits the oocyte to develop to metaphase II, and $\beta$-ME reverses such inhibitory effects. The cooperation of gonadotropins and $\beta$-ME stimulates the meiotic resumption and especially, promotes the CEOs to develop to metaphase II in both models. Moreover, HX might contribute to the fragility of oocyte zona pellucida and gonadotropins, nitric oxide and $\beta$-ME could alleviate it separately, and cooperatively. It is concluded that NO exerts different functions in two models and $\beta$-ME affected the functions of NO in different models.

• Journal of Embryo Transfer
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• 제21권3호
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• pp.255-262
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• 2006

The purpose of this study is to investigate the effects of vitrification in open pulled straws (OPS) methods on in vitro survival ability of porcine embryos. For in vitro maturation of immature oocytes, the porcine ovaries were collected from local slaughter-house. The cumulus-oocytes complexes were aspirated from 2 to 6 mm follicles. The collected oocytes were cultured for in vitro maturation in NCSU-23 medium with 5 mM hypotaurine, 0.57 mM cysteine, 10% porcine follicle fluid, 10 IU/ml PMSG and 10 IU/ml hCG for $21{\sim}22$ hrs. Then, the oocytes were more cultured $21{\sim}22$ hrs in vitro maturation in medium removed hormones. The frozen-thawed spermatozoa were washed by centrifugation 2 times for 10 min at 1,500 rpm in D-PBS with 5.56 mM glucose, 0.33 mM Na-pyruvate, 100 IU/ml penicillin, $100 {\mu}g/ml$ streptomycin and 4 mg/ml BSA. The fertilization medium used mTBM with 2 mM caffeine and 2 mg/ml BSA and adjusted to a pH of 7.2 to 7.4. The final concentration of spermatozoa was adjusted to $2.5{\times}10^6$cells/ml motile sperm during fertilization in vitro. At 8 hrs after insemination, the oocytes were transferred into NCSU-23 medium with 5.0 mM hypotaurine, 4 mg/ml BSA and 10 ng/ml EGF and cultured for 7 days. When the blastocysts of different stages were frozen-thawed by OPS methods, the proportions of embryos with normal morphology were significantly (p<0.05) higher in embryos frozen-thawed at expanded blastocyst stage (38.9%) than in early blastocyst stage (28.3%). On the other hand, the proportions of embryos damaged after frozen-thawing were significantly (p<0.05) higher in embryos frozen at early blastocyst stages than in expanded blastocyst stage. In another experiment, the normal embryos morphology after frozen- thawing were further cultured for 48 hrs. After culture, the proportions of embryos hatched were 6.7, 20.0 and 33.3% for embryos frozen-thawed at early blastocyst, mid-blastocyst and expanded blastocyst stages. These finding indicate the possible broader application for OPS methods, as frozen-thawed embryos may be accompanied by developmental stage according to requirements of the survival ability after freezing of blastocyst stage in the pig.