• Journal of Embryo Transfer
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• v.17 no.2
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• pp.145-151
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• 2002

The aim of these experiments was to investigate in vitro nuclear maturation of canine oocyte collected from various stages of estrus cycles, Ovaries were obtained from 1 to 4 year-old mongrel bitch and minced for oocyte collection in phosphate buffered saline with 100 iu penicillin-G $m\ell$／sup -1／, 50 $\mu\textrm{g}$ streptomycin sulphate $m\ell$／sup -1／ and 0.1％ (w／v) polyvinyl alcohol. Cumulus-oocyte-complexes (COCs) were washed in HEPES buffered tissue culture medium (TCM)199 and in vitro matured in TCM-199 culture medium supplemented with sodium pyruvate 0.028mg/$m\ell$, L-glutamine 0.146mg／$m\ell$, penicillin G 10,000 IU／$m\ell$, streptomycin 0.031mg／$m\ell$ and 10％ (v／v) fatal calf serum. COCs were in vitro matured for 48～72 hrs at 39$^{\circ}C$ in humidified 5％ $CO_2$ in air atmosphere. In vitro matured oocytes were remove the cumulus cells using 0.2％ (v／v) hyaluronidase. After denuding, oocyte were placed in acetic acid : methanol : chlorform solusion (3:6 : 1.5 v／v) for 30 sec and acetic acid: ethanol(1:3 v／v) for 48hrs fixation. Nuclear maturation was classified to GV, GVBD, MI, MII and degenerate oocyte under microscopy after 1％ aceto-orcein stain. In vitro maturation rates at 48hrs were not significantly difference among the oocytes collected from different stage of estrus at 15.9％, 16.3％, 23.7％ and 18.2％ for anestrus, proestrus, estrus and diestrus. However, the oocytes maturation(36.6％) of collected from estrus ovaries were significantly different from oocytes derived from proesturs, diestrus and anestrus ovaries(30.8％, 17.5％ and 22.1％; p＜0.05). The overall in vitro maturation rates were significantly higher (p＜0.05) in 72hrs culture than 48hrs culture system. In summary, there was a tendency for higher in vitro maturation rates with the oocyte collected from estrus ovary than other stages of estrus. Also, for nuclear maturation, in vitro culture of oocyte for 72hrs was better than 48hrs culture.

• Korean Journal of Animal Reproduction
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• v.14 no.1
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• pp.57-65
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• 1990

These experiment were carried out ; (1) to investigate the changes of progesterone in plasma, whole milk and skim milk during oestrus cycle and pregnancy and ; (20) to evaluate a chemiluminescence Immunoassay(CIA) as an alternative method by measuring the progesterone concentration is skim milk by RIA and CIA. The results obtained in these experiments were summerized as follows ; 1. Plasma progesterone levels in non-pregnant Holstein during oestrus cycle were relatively low until day 8 after oestrus. And then, the progesterone level began to increase and reached a peak with 6.3ng/ml on day 14 and then declined repidly to 1.5ng/ml and 2.2ng/ml on day 18 and 20, respectively. 2. Whole milk progesterone level in pregnant Holstein increased from 1.0ng/ml on oestrus to 16.0ng/ml on day 8 and then remained from 11.0ng/ml on day 10 to 22.0ng/ml on day 22. 3. In non-pregnant Holstein, whole milk pregesterone lev디 was 1.5ng/ml on oestrus and began to increase rapidly from day 6 after oestrus and exhibited a ranged of levels, 17.8~20.0ng/ml from day 6 to day 16 after oestrus. 4. Skim milk progesterone levels in pregnant Holstein were a range of 130~490pg/ml at the time of estrus and began to increase continually till then showing constant levels ranging from 1300pg/ml on day 10 to 1650pg/ml on day 22. 5. In non-pregnant Holstein, skim milk progesterone level was 160pg/ml on oestrus and began to increase from 190pg/ml on day 2 after oestrus to day 8 and then keep constant levels ran ging from 1050 to 1300pg/ml from day 8 to day 16 and then decreased to 240~450pg/ml from day 18 to day 22 after oestrus. 6. The results obtained from CIA for the analysis of skim milk progesterone were in good agreement with the values derived by RIA.(r=0.914)

• Korean Journal of Animal Reproduction
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• v.25 no.2
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• pp.93-100
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• 2001

This study was conducted to evaluate the nuclear development of preantral follicle oocytes in dog following collecting from different estrus ovaries and oocyte diameter. To do this, ovaries were collected from Sunchon livestock station by ovarioectomy and then transported to laboratory in 3$0^{\circ}C$ saline within 2 h. All of the ovaries were washed three times with saline supplemented with 100 IU Penicillin and 100 $\mu\textrm{g}$/$m\ell$ Streptomycin and then sliced with blade in 100 mm dish. All of the oocytes collected were classified the oocyte size such as over 110 ${\mu}{\textrm}{m}$ or under 110 ${\mu}{\textrm}{m}$ and the estrus status. To induce the nuclear development, oocytes were cultured in TCM199 $\alpha$-MEM supplemented with 10% FCS, 0.1 $\mu\textrm{g}$/$m\ell$ sodium pyruvate, 100 ng/$m\ell$ FSH, 100 ng/$m\ell$ EGF, 1% ITS, 100 IU/$m\ell$ penicillin, 100 $\mu\textrm{g}$/$m\ell$ streptomycin at 38.5$^{\circ}C$, 5% $CO_2$ incubator for 72 h. After culture, all of the oocytes were stained in 1% orcein following fixed in 45% Acetic acid for 48 h. The oocyte recovery rates of over 110 ${\mu}{\textrm}{m}$ from estrus ovary (63.6%) were significantly higher rather than that in anestrus status (51.5%). Oocyte recovery rate per ovary of over 110 ${\mu}{\textrm}{m}$ in estrus status ovary (22.6/63.8%) was also significantly higher rather than that in anestrus status ovary (8.2/51.6%). Nuclear development to MI of over 110 ${\mu}{\textrm}{m}$ in estrus ovary (24.3%) was significantly higher rather than those in under 110 ${\mu}{\textrm}{m}$ or over and under 110 ${\mu}{\textrm}{m}$ in anestrus (2.5, 6.8 and 0.0%), respectively (P ＜ 0.05). Nuclear development to AT and MII of over 110 ${\mu}{\textrm}{m}$ in estrus was more developed in other groups. Nuclear development to MI in TCM199 (21.8%) was significantly higher than in $\alpha$-MEM (10.0%). Altho $\mu\textrm{g}$h the development rate to AT was significantly different between TCM199 (7.3%) and $\alpha$-MEM (1.1%), but to MII was not different between TCM199 (0.9%) and $\alpha$-MEM (1.1%). The results indicated that over 110 ${\mu}{\textrm}{m}$ oocytes was could be recovery from estrus status ovary, bigger oocytes were more developed to MI, AT or MII in TCM199.

• Proceedings of the KSAR Conference
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• 2004.06a
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• pp.293-293
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• 2004

본 실험은 도축전 암소에 성선자극호르몬(Gonadon, 동방(주) : 3 ㎖ 목근육 주사, Gonadorelin 100 ㎍/㎖)을 처리하여 인위적으로 새로운 발정주기를 유도하여 난자 회수율 및 수정란 생산성을 향상시키고자 실시하였다. ① 실험대조군으로 85두(T1), 호르몬처리 37두를 대상으로 처리시간(T2 ; 72hrs, T3 ; 96 hrs, T4 ; 120. T5 ; 148 hrs.)에 따라 도축 후 난소로부터 혈통수정란 생산을 위해 개체별로 난자 회수량을 확인하였다. 회수된 난자를 이용하여 체외성숙 및 체외수정을 실시하였고, 이들 난자의 발육상태, 난발달 및 동결가능 수정란수를 각각 확인하였다. (중략)

• Proceedings of the KSAR Conference
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• 2002.06a
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• pp.57-57
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• 2002

멸종위기에 있는 우리나라 재래돼지의 안전한 보존을 위해서는 수정란 동결보존이 필수적이며 동결에 적합한 수정란 생산이 선행되어야 한다. PMSG/hCG 에 의한 돼지 체내수정란 생산 연구는 주로 대형종을 중심으로 이루어져 왔으며 우리나라 재래돼지와 같은 중소형종에 대한 연구는 미미한 실정이다. 본 연구는 다배란를 유도하기 위해 투여하는 PMSG/hCG 에 대한 우리나라 재래돼지의 난소반응과 외과적 채란에 의한 수정란 회수율을 구명함으로써 재래돼지 체내수정란 생산의 효율성을 높이기 위해 수행되었다. 18일 동안 20㎎/day의 Altrenogest(Regumate/sup R/, Intervert)로 발정주기를 동기화 시킨 미경산 재래돼지 11두를 3군으로 나누어 각각 500, 750, 1,000 IU의 PMSG(Folligon/sup R/, Intervert)를 주사하고 80-84시간 후 500 IU의 hCG(Chorulon/sup R/, Intervert)를 주사하여 다배란을 유도하였다. (중략)

• Korean Journal of Animal Reproduction
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• v.26 no.2
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• pp.193-199
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• 2002

This study was conducted to investigate the optimal physiologic mating time in Hanwoo for protection to decrease of reproductivity and improvement of production of offspring. We observed 32 cows that were devide into 4 parts of treatment : T1(12 months of age and 0.5kg daily gain), T2(12 months of age and 0.8kg daily gain), T3(15 months of age and 0.5kg daily gain) and T4(18 months and 0.5kg daily gain). The first heat of treated cows was 263.3$\pm$6.4 days and average weight was 181.1$\pm$11.3kg. It was revealed the conception rates of first insemination were 25%(T1), 75%(T4) and number of insemination of T3 and T4(both 1.5) was lower than T1 and T2(2.3 and 2.4). In return of estrus after heifer's first parturition, they(T1, T2, T3 and T4) showed 66.2 days, 76.7 days, 62.4 days and 68.5 days respectively and the average was 65.7 days. Plasma progesterone(P4) concentration was nearly the same during the observation periods of treated cows and P4 was released just after 12 months. Only 5 cows (15.6%) in 32 were showed normal estrus cycle and ovulation before 12 months. Before and after parturition, P4 concentration was decreased fastly and then there was no detection of P4 from after parturition to 40 days after milking. P4 would be released again on 45 day after parturition. The results were summarized as that the optimal mating time of Hanwoo heifers was decided by the 14 months of age, 110 cm height and 265kg weight.

• Journal of Practical Agriculture & Fisheries Research
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• v.4 no.1
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• pp.110-118
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• 2002

This study was conducted to investigate the physiology of optimal reproductive age in Hanwoo for protection to decrease of reproductivity and improvement of production of offspring. Thirty two cows were devided into 4 groups of treatments : T1(12 months of age and 0.5kg daily gain), T2(12 months of age and 0.8kg daily gain), T3(15 months of age and 0.5kg daily gain) and T4(18 months of age and 0.5kg daily gain). The days of the first heat of treated cows were 263.3±6.4 days and average weight was 181.1±11.3kg. The conception rates of first insemination were 25%(T1) and 75%(T4), and the number of insemination of T3 and T4(both 1.5) was lower than those of T1 and T2(2.3 and 2.4), respectively. With regard to estrus return after the first parturition, T1, T2, T3 and T4 showed 66.2,76.7, 62.4 and 68.5 days, respectively, indicating the average days of estrus return was 65.7. Plasma progesterone(P4) concentration was nearly the same during the observation periods of treated cows and P4 began to be detected after 12months. Only 5(15.6%) out of 32cows showed normal estrus cycle and ovulation before 12 months. During the peri-parturition period, P4 concentration was rapidly decreased and there was no detection of P4 from parturition to 40 days after milking. P4 would be released again on 45 day after parturition. The results imply that the optimal reproductive age of Hanwoo heifers would be around at the 14 months of age, 110cm height and 265kg weight.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.126-126
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• 2003

유전적으로 우수한 미경산우를 선발하여 조기에 우수한 수정란을 생산하여 이식 후 개량을 촉진한다는 것은 소의 육종개량에서 매우 중요한 일이다. 일반적으로 경산우에 비해 미경산우는 수정란의 생산효율이 떨어진다는 보고가 있다. 최근에는 다배란 처리시 FSH와 Estradiol Benzoate(EB)를 사용하여 다배란처리 효율을 개선하였다는 보고(Matoba 등, 2002)가 있어 EB의 첨가가 미경산우에 있어 수정란 생산 효율을 개선하는 지를 조사하기 위해 60일 간격으로 반복하여 다배란 유기실험을 수행하였다. 공란우는 11 -15개월령의 한우 16두를 평균 90일 주기로 반복 다배란 처리우로 사용하였으며 발정 주기중 황체 중기 소에게 CIDR plus를 질내에 삽입하고 다음날 EB를 근육주사하였으며, 120시간 후 부터 FSH 50mg을 1일 2회, 4일간 주사하고 마지막 날 prostaglandin제제를 25mg 근육주사한 다음에 48, 60시간 후 인공수정을 반복 실시하였다. 인공 수정시는 배란을 촉진하기 위해 GnRH제(콘세랄)을 50mg을 근육주사하였다. (중략)

• Applied Microscopy
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• v.12 no.1
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• pp.57-68
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• 1982

Cyclical changes in the fine structures of endometrial stroma of guinea pig during the estrous cycle were studied by electron microscopy. Cytochemical studies were made in order to investigate the ultrastructural localization of the acid phosphatase,alkaline phosphatase and ATPase in these cells. The results obtained are as follows: 1. During estrus collagen fibers were most abundant in the stroma. The stromal cells showed increases in the number of several cytoplasmic organelles, especially the rough endoplasmic reticulum was significantly increased and the structures were greatly differentiated. 2. Many cytoplasmic processes and cell debris have been distributed in the stroma of metestrus. The distributions were increased and degenerated mitochondria were observed during diestrus. 3. Cytochemical studies indicated that during metestrus and diestrus acid phosphatase activities were localized in the degenerating collagen fibers. Alkaline phosphatase activities were weak in the collegen fibers during proestrus and estrus which intense activities were localized around the cell membrane during metestrus and diestrus. ATPase activities were present on the cell membrane and intercellular space of stromal cell during proestrus and estrus.

• Journal of Veterinary Clinics
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• v.14 no.1
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• pp.93-98
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• 1997

The vaginal endoscopy was performed in two Labrador Retrievers from onset of vaginal bleeding until metestrus accompanying with vaginal smear and blood progesterone concentration to investigate estral changes, and the intrauterine infusion of radiopaque fluid using vaginal endoscope (rigid 25-degree endoscope, Karl Storz) was detected by radiogrphy to determine if theis technique is feasible for future artificial insemination in the dogs. During the proestrus vaginal mucous membrane folds were edematous and a large amount of clear red discharge was present. In the beginning of estrus, the mucous membrane folds was present and the amount of the red discharge was decreased. In the estrus there were obbvious shrinkage, angulation of the membrane folds and decreasing of the discharge. In metestrus the vaginal folds started to round out and the surface appeared moist and sticky. The endoscope used for endoscopic monitoring of the vaginal mucosa was advanced until the external os of the cervix could be visualized. A plastic catheter, 8Fr guage was manupulated into the cervical os and then advanced through into the uterus. The radiopaque fluid was injected into the uterus through the catheter and radiographs taken to detect the site of insemination using the endoscopic technique revealed the exact infusion of the fluid in the uterus, consequentlyto prove that this technique is usable for more successful artificial insemintion tn the bitch.