• 대한수의학회지
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• 제40권4호
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• pp.769-775
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• 2000

This study was investigated the ovarian response following superovulation treatment at different day of diestrus. The criterion for the presence or absence of a dominant follicle based on their morphological examination. Dominant follicle was puntured 48 hrs before the oneset of superovulation treatment by ultrasonography guided aspiration needle. Superovulation was induced by subcutaneous administration of FSH twice a day for 4 day in a decreasing regimen. There was no significant different between presence of dominont follicle and progesterone concentration/diameter of corpus luteum in HanWoo. Number of corpus luteum of donor after superovulation treatment was not significantly different in FSH administration at day 9, 11 and day 13 of estrus($14.5{\pm}4.5$, $15.5{\pm}5.6$ and $11.0{\pm}5.5$, respectively). But, the diameter of CL was significantly correlate(R2 = 0.757) with progesterone levels on day of superovulatory induction. After 7 days of artificial insemination, the embryos at 7 days were collected by uterine flushing after dominant follicle aspiration and superovulation treatment, and evaluated their quality by morphological criteria. Fifty five embryos with excellent, good and fair grade were transferred into 24 recipient cows. Seventeen offsprings, 1 of triplet, 4 of twins and 6 of singlet, were yield from 10 recipient cow. In conclusion, the present study showed that 1) dominant follicle can be determined by ultrasonography with rectal palpation by morphological evaluations, 2) superovulation response after follicular aspiration was not differ at day 9, 11 and 13 of estrus, 3) dominant follicle did not affect to progesterone concentration and diameter of CL, and 4) diameter of CL was significantly correlate to the level of progesterone concentrations in HanWoo.

• 한국수정란이식학회지
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• 제32권3호
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• pp.147-151
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• 2017

Commercial applications of OPU/IVP were to produce embryos and calves from high genetic cows. The aim of this present study was to compare the number of recovered oocytes and cultured In vitro produced embryos from Ovum Pick-up (OPU). OPU derived embryo production was carried out of oocytes by ultrasonographic guided follicular aspiration and then produced in vitro produced blastocysts by IVP culture system. In result, the rate of recovered oocytes was obtained 612 (57.2%) and 451(73.7) G1+G2 grade oocytes. No difference of recovered rate (51.1~62.1%) was seen in six donor. The rate of cleavage and blastocyst development were obtained 320 (70.9%) and 78 (24.4%) that was $3.3{\pm}0.4$ cleaved embryo and $0.9{\pm}0.2$ blastocysts per session. Cleavage rate of OPU oocytes in No. 6 donor was 90.6%, significantly (P<0.05) higher than that in the other donors, However, blastocysts was similar (25.8~30.0%). In conclusion, limited numbers of OPU oocytes had competent development when cultured in SOF culture medium.

• Reproductive and Developmental Biology
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• 제35권3호
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• pp.369-375
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• 2011

본 연구는 한우의 체외수정란 생산에서 체외성숙, 체외 배양시 배양액에 항산화제인 ${\alpha}$-Tocopherol의 첨가 농도에 따른 대조구와 처리구 난포란의 발달과 배반포 발생율 및 초자화 동결 후 생존성에 미치는 영향을 구명하고자 실험을 실시하였다. ${\alpha}$-Tocopherol 첨가 농도에 따른 분할율에서는 7~10월까지 비교한 결과, ${\alpha}$-tocopherol 200 ${\mu}M$ 첨가된 처리구가 대조구 및 타 처리구보다 높은 경향을 보였으나, 유의적 차이는 없었다. 배반포 발생율에서는 8~10월에서 ${\alpha}$-tocopherol 200 ${\mu}M$ 첨가된 처리구 $38.60{\pm}7.12%$가 대조구 및 타 처리구들보다 유의적으로 높게 (p<0.05) 나타났다. 초자화 동결 전 후의 총세포수는 ${\alpha}$-tocopherol 200 ${\mu}M$ 처리구에서 $115.80{\pm}6.61$개, $106.33{\pm}3.50$개로서, 다른 대조구 및 타 처리구보다 높은 세포수를 보였다. 월별에 따른 수정란의 배반포 발생율 및 동결 전후의 총세포수에서는 7~8월이 9~10월보다 유의적으로 높게 (p<0.05) 나타났다.

• 한국가축번식학회지
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• 제26권2호
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• pp.193-199
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• 2002

본 연구는 한우의 생리적인 번식적령기를 구명하는 기초자료를 확보하여 조기번식에 따른 번식능력의 감소를 방지하고 한우 암소의 번식능력을 증진하여 농가의 소득증대에 이바지할 목적으로 각 처리구당 4구로 나누어 총 32두를 공시하여 얻어진 연구결과는 다음과 같다. 분만후 송아지의 평균 생시 체중은 전 구간에서 평균 23.5$\pm$0.6kg으로 나타났으며 암송아지는 21.8$\pm$0.8kg에 비해 수송아지는 평균 25.0 $\pm$0.8kg으로 다소 높게 나타났다. 또한 송아지 성장단계별 체중은 T2구에서 유의적인 증가 추세를 보였다. 따라서 송아지의 체중변화는 어미소의 영양상태에 따라 중요하게 작용하는 것으로 나타났다. 초발정일령은 전 구간에서 평균일령과 평균체중은 각각 263.3$\pm$6.4일과 181.1$\pm$11.3일로 나타났다. 1회째 인공수정에 의한 수태율은 T1구에서 25%로 가장 낮았으며 T4구에는 75.0%로 가장 높았다. 수태당 종부회수는 T1와 T2구의 2.3, 2.4회에 비해 T3, T4구가 1.5회로 낮았다. 초임 월령별 분만후 발정재귀일수는 T1구에서는 평균 66.2일, T2구에서는 76.7일, T3구에서는 62.4일, T4구에서는 68.5일이었으며 전 구간의 평균 발정재귀일수는 65.7일로 나타났다. 초발정 전후의 혈중 progesterone 농도는 전 구간에서 거의 유사한 결과를 보였다. 즉, 혈중 progesterone 분비는 생후 12개월령이 지나야만이 전 개체에서 정상적인 호르몬 분비상태가 나타나고 있다. 본 연구의 결과로는 12개월 이전에 난포의 배란과 정상 발정주기가 반복되는 개체는 총 32두중 5두만(15.6%)으로 확인되었다. 임신기간중 혈중 progesterone 농도는 임신 중기 이후에 비해 착상과 태반형성이 이루어지는 시기인 임신 초기에 다소 높은 경향이었으며 임신중기에는 다소 감소되었다. 분만 전후 혈중 progesterone 농도는 전 구간에서 분만 7일전부터 급격하게 감소되었으며 분만후부터 포유 약40일까지 progesterone이 거의 검출되지 않았으며 분만후 35일부터 본격적으로 progesterone이 분비되고 있음이 확인되었다. 따라서 분만후 정상 발정주기의 반복은 분만후 약 40일 이후 비로소 시작되는 것으로 사료된다. 이상으로 본 연구의 결과를 요약하여 한우의 초임유도시 최적 초임시기의 결정요소는 14개월령과 체고 약 110cm, 체중 약 265kg 정도의 3가지 요건을 갖춘 후 초임유도를 시키는 것이 바람직하다고 사료된다.

• 한국수정란이식학회지
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• 제24권1호
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• pp.57-64
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• 2009

본 연구는 한우의 난포낭종 및 황체낭종에서 세포자멸사 관련 유전자의 발현 변화를 조사함으로써 난소낭종과 세포자 멸사간의 관계를 정리하고자 한다. 마이크로어레이 분석 결과, 난포낭종에서 PIK3R2와 AKT1가 유의하게 증가하였고, 황체낭종에서는 증가되는 세포자멸사 관련 DEGs, TNF-RAF2, PRLR, FOXL2, STK4 및 COL4A3와 감소하는 DEGs, INHA, CIDEB, BCL10 및 FASLG가 포함되었다. 정량적 역전사 중합 효소 반응을 이용하여 mRNA 발현 변화를 검증한 결과 INHA, CIDEB, BCL10 및 FASLG만 마이크로어레이와 동일한 결과를 보였다. INHA의 경우 마이크로어레이 결과에서는 1.43배 감소하고 정량적 역전사 중합효소 반응 결과에서는 12.3배 감소하였다. 세포사멸 확인을 위하여 TUNEL 형광염색과 ERK, JNK, p38, PI3K 및 Akt/PKB의 웨스턴 블랏 분석을 실시한 결과 정상난소와의 유의한 차이를 확인할 수 없었다. Caspase의 활성 및 Bax, Bcl-2, Bcl-xL의 단백질 분석 역시 정상난소와 유의한 차이를 보이지 않았다. 오히려 PI3K와 p-Akt의 발현이 정상에 비하여 감소하는 경향을 보였다. 이러한 결과들로 미루어 보아 난소낭종 시 볼 수 있는 세포자멸사의 낮은 발현율은 세포 사멸인자의 발현 감소로 나타날 수 있을 것이라 생각된다.

• 한국가축번식학회지
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• 제11권1호
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• pp.73-84
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• 1987

This experiment was carried out to investigate the effects of hormone addition(FSH, HCG, estrogen and progesterone) and composition (BSA and FCS) of mKRB on the in vitro maturation and fertilizability of follicular oocytes of the Korean native cattle. The ovaries were removed at a slaughterhouse, returned to laboratory in a thermostat (30-35$^{\circ}C$) within 4 hr, and collected by aspirating normal follicles which had diameters of 1 to 6 mm. The oocytes with cumulus cells were cultured for 8, 16, 24 and 30 hr in a modified KRB solution containing BSA or FCS and hormones. The in vitro matured oocytes in mKRB containing FCS, FSH and steroids were transferred in the rabbit uterus for examination of their in vivo fertilizability with bovine sperm preincubated 4 to 6 hr in the rabbit uterus. 1. The mean number of oocytes collected per cattle was 6.5 from 1-3mm follicles, 1.3 from 4-6mm follicles, and total was 7.7. 2. The meiotic division at 16hr-cuture in the oocytes from 1-3mm follicles was slightly stimulated by the addition of FSH in mKRB + BSA solution compared with the control. At 30hr-culture, their maturation rates(%Met II) were also increased by FSH of 1 $\mu\textrm{g}$/ml(38.4%) and 5$\mu\textrm{g}$/ml(35.7%) as compared with the control (21.4%). The maturation rate at 30hr-culture in the oocytes from 4-6mm follicles was 53.8% and 57.1% by the FSH addition of 1$\mu\textrm{g}$/ml and 5$\mu\textrm{g}$/ml, respectively. These rates were similar with the control(57.1%), but higher than those of oocytes from 1-3mm follicles. 3. The meiotic division at 16hr-culture in the oocytes from 1-3mm follicles was stimulated by the HCG addition of 1IU/ml and 5IU/ml. However, the maturation rate at 30hr-culture was greatly decreased by the HCG addtion (26.6% and 13.3%) compared with the control(53.3%) and these rates (30.8%) in the oocytes from 4-6mm follicles were also lower than that fo the control(58.3%). 4. Low maturation rate (37.5%) of the oocytes cultured in mKRB containing BSA and 5IU/ml HCG was increased (55.0%) when 15% FCS with HCG was added to mKRB instead of BSA. 5. When 16hr-cultured oocytes in mKRB containing BSA and gonadotropins (5$\mu\textrm{g}$/ml FSH and 5IU/ml HCG) were transferred in the medium without gonadotropins and recultured for 16hr, the maturation rate of HCG-treated oocytes was greatly improved. 6. The maturation rates of oocytes were greatly affected by steroids. The combined addition of FCS+FSH+estrogen or +progesterone to mKRB increased the maturation rate compared with the combination of BSA+FSH or FCS+FSH in mKRB. 7. The fertilization rate, presence of pronuclei, was increased by the combination of FCS+FSH+p in mKRB as compared with that (5.6%) of BSA+FSH and the rates of FCS+FSH+steroids ranged from 12.5 to 17.6%.

• 한국수정란이식학회지
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• 제27권3호
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• pp.141-147
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• 2012

This study was operated to establish induction using ultrasonography by estimating the relation of follicle size and estrus manifestation. Clinical estrus symptoms were observed 97.4% in cows and 87.5% in heifers when overall 55 cows were induced to estrus in a single dose of $PGF_{2{\alpha}}$ after verifying CL through ultrasonography, which means estrus hours among those 52 cows showing the clinical estrus symptoms were estimated 2.39 days on cows and for 2.37 days on heifers which showed no differences (p>0.05). The estrus manifestation hours according to the follicle size in cows didn't have any significance each other (p>0.05), though estrus hours was 54 hours (the shortest) with follicle size bigger than 10 mm and were made up within 69 hours. The estrus manifestation hours according to the follicle size in heifers didn't have any significance each other (p>0.05) and took around 42 hours (the shortest) with follicle size of 5mm (the smallest) and were made up within 66 hours. Follicles after $PGF_{2{\alpha}}$ injection were ovulated and assigned to many phases as follows; Group 1 (growing phase) - continuously growing into ovulation, Group 2 (growing and static phase) - delaying in growth after the growth of follicles, Group 3 (static and growing phase) - growing after growth delay, Group 4 (regressing and new growing phase) - the follicle is closed and a new follicle grows. In addition, the process of follicle development and estrus hours had no significance each other (p>0.05), though estrus manifestation hours in Group 1 and 2 was relatively short, and in Group 3 and 4 for a relatively long time. In the result of all above, the estrus manifestation hours after $PGF_{2{\alpha}}$ injection has no differences accoring to the follicle size in cows and heifers. Therefore, High pregnancy rate is obtained when practicing artificial insemination within 3 days in estrus or TAI in 72 to 80 hours after adminitrating $PGF_{2{\alpha}}$.

• 대한수의학회지
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• 제36권4호
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• pp.929-939
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• 1996

The present study was undertaken to establish a relationship between bovine follicle size and oocyte diameter, compare the nuclear maturation competence of oocytes of different diameter groups and the nuclear maturation changes in Korean Native Cattle according to in vitro maturation period. To compare the relationship between follicle size and oocyte diameter, follicles were dissected, measured, and assigned to one of the following size categories($4{\geq}mm$, 3-4mm, 2-3mm, 1-2mm, and < 1mm), investigate the maturation competence in the different-sized oocytes, which were divided into three groups( < $110{\mu}m$, 110 - < $120{\mu}m$, and ${\geq}120{\mu}m$). Oocytes were cultured in the culture medium during 0, 6, 12, 18, and 24hrs, respectively, stained, and measured the nuclear maturation degree according to period. When compared the relationship between follicle size and intrafollicular oocyte diameter, oocyte diameters of three groups of ${\geq}3mm$ follicle-sized were significantly higher than < 3mm (p<0.01). After in vitro maturation, the rates reached to MI stage of < $110{\mu}m$ oocyte groups(25%) was higher than $110-120{\mu}m$ and ${\geq}120{\mu}m$ oocyte groups(11 and 10%) reached to the same stage(p<0.01), and the rates throughout MII stage of $110-120{\mu}m$ and ${\geq}120{\mu}m$ and < $110{\mu}m$(70 and 76%) groups were higher than < $110{\mu}m$(35%)(p<0.01). When nuclear maturation rates were measured according to period, < 6hr groups(7 and 10%) showed lower rates reached to MI than ${\geq}12hr$ groups(100%), 24hr groups(76%) revealed higher rates throughout MII than 18hr groups(40%). These results indicate that the preparation of oocyte for the production of in vitro fertilization embryos and nuclear transplantation ones could be adapted, as follicle increased up to appointed size there was a corresponding increase in oocyte diameter, and differences of nuclear maturation rate revealed according to oocyte diameter and maturation period.