• 한국수정란이식학회지
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• 제27권4호
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• pp.237-243
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• 2012

This research was investigated the relationship between the number of the transferable embryos and estrus expression rate, BCS (Body Condition Score), which affect the nutritional state of the cow, in Holstein donor cows. CIDRs were inserted into the vaginas of twenty two head of Holstein cows, regardless of estrous cycle. Superovulation was induced using folliclar stimulating hormone (FSH). For artificial insemination, donor cows were injected with $PGF_{2{\alpha}}$ and estrus was checked about 48 hours after the injection. Then they were treated with 4 straws of semen 3 times, with 12-hour intervals. Embryos were collected by a non-surgical method 7 days after the first artificial insemination. When BCS was $$\leq_-$$2.5, the total number of collected ova was 7.3 + 1.9, which is significantly lower (p<0.05) than the numbers 15.4 + 2.8 and 15.4 + 2.1 that were obtained when BCSs were 2.75 and $$\geq_-$$3.0, respectively. Whereas the numbers of transferable embryos were 5.2 + 1.4 when BCS was $$\leq_-$$2.5, which was smaller than the numbers 6.0 + 2.1 and 8.5 + 1.8 that were obtained when BCSs were 2.75 and $$\geq_-$$3.0, respectively; however, the differences were not significant. As for estrus induction rate, the cow groups whose BCSs were 2.75 and $$\geq_-$$3.0 showed 100.0% and 95.0%, respectively. Whereas the cow group whose BCS was $$\leq_-$$2.5 showed 57.1%, and the differences were significant (p< 0.05). As for estrous expression rate, the cow groups whose BCSs were $$\leq_-$$2.5, 2.75 and $$\geq_-$$3.0 showed 100.0%, 100.0% and 85.7%, respectively; however, the differences were not significant. According to the result of this research, it is considered that the total number of collected ova and the number of transferable embryos will be affected by the nutritional state before and after in vivo embryo production and superovulation treatment, and that although the mechanism is not clear, poor stockbreeding management and nutritional level would cause the decrease of ovum recovery rate and the number of transferable embryos in high-producing cows. On the other hand, diverse researches on the superovulation treatment method that is suitable for high-producing Holstein donor cows would contribute to preventing ovarian cyclicity disorder, as well as to the early multiplication of cows with superior genes by increasing the utilization value of donor cows.

• The Korean Journal of Physiology and Pharmacology
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• 제1권6호
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• pp.759-767
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• 1997

In the present study, we characterized the non-vascular smooth muscle relaxant effects of a novel benzoyran derivative ,SKP-450 (2-[2'(1',3'-dioxolone)-2-methyl-4- (2'-oxo-1'-pyrrolidinyl) -6-nitro-2H-1- benzopyran) and its metabolite, SKP-310, in comparison with levcromakalim (LCRK). In the rat stomach fundus, the spontaneous motility stimulated by $10^{-6.5}\;M$ bethanechol was completely eliminated not only by $10^{-7}\;M$ SKP-450 but also by $10^{-6}\;M$ LCRK, which were blocked by $10^{-6}\;M$ glibenclamide. The inhibitory effect of SKP-450 $pD_2,\;3.94{\pm}0.66)$ was much less than LCRK $(pD2,\;5.73{\pm}0.38,\;p<0.05)$. In the bethanechol $(10{-6.5 }\;M)-stimulated$ urinary bladder, the tonus was decreased in association with elimination of spontaneous motility by $10^{-7}\;M$ M SKP-450 and $10^{-6}\;M\;LCRK\;(pD2,\;6.77{\pm}0.06)\;(P<0.05)$, which were inhibitable by $10^{-6}\;M$ glibenclamide. The inhibitory effect of SKP-450 $(pD2,\;7.66{\pm}0.05)$ was significantly more potent than that of LCRK $(pD2,\;6.77{\pm}0.06,\;p<0.05)$. In the rat uterus stimulated by $PGF_{2\alpha}\;(10^{-7}\;M)$, both increased tonus and spontaneous motility were eliminated by $10^{-6}\;M$ LCRK with slight depression of the tonus, but not by SKP-450 $(10^{-5}\;M)$. The stimulated trachea of guinea-pig by $10^{-6.5}\;M$ bethanechol was moderately suppressed by SKP-450 $(10^{-6}{sim}10^{-5}\;M)$ but little by SKP-310. In association with the relaxant effects, SKP-450 $(10^{-6}\;M)$ and LCRK $(10^{-5}\;M)$ caused a significant stimulation of the $^{86}Rb$ efflux from rat urinary bladder and stomach fundus, which were antagonized by $10^{-5}\;M$ glibenclamide, whereas the $K^+$ channel openers could not exert a stimulation of the $^{86}Rb$ efflux from rat uterus. In conclusion, it is suggested that SKP-450 exerts potent relaxant effects on the urinary bladder detrusor muscle and duodenum, whereas it shows much less effect on stomach fundus and uterus as contrasted to LCRK.

• 한국수정란이식학회지
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• 제26권3호
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• pp.159-164
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• 2011

This study was performed in order to determine optimum flushing solution using the direct embryo collection (DEC). Donors, at random stages of the estrous cycle, received a CIDR. 7 days later, 200 mg FSH was treated with 40, 30, 20, 10 mg FSH levels in declining doses twice daily by intramuscular injection for 4 days. On the 3$^{rd}$ day administration of FSH, 25 mg $PGF_2{\alpha}$ was administered and CIDR was withdrawn. After FSH injections were complete, donors were artificially inseminated twice at 12 hr intervals. The donor cattle received 250 ${\mu}g$ GnRH at time of 1$^{st}$ insemination and embryos were recovered 8 days after the 1$^{st}$ insemination. Embryo collection from superovulated donors were performed to flushing by DEC and conventional method. As a results, the average number of recovered embryos were significantly higher as 19.1${\pm}$1.40 with DEC method than 12.0${\pm}$0.44 with conventional embryo collection method, respectively (p<0.05). Also, The average number of transferable embryos were significantly higher (p<0.05) as 15.8${\pm}$1.72 with DEC method than 6.9${\pm}$0.35 from conventional embryo recovery procedures. Meanwhile, number of recovered embryos and number of recovered transferable embryos following the number of flushing times until 6${dr}$ flushing were significantly higher as 8.6${\pm}$0.53 and 8.6${\pm}$0.53 from 2$^{nd}$ flushing time than other groups (p<0.05). No. of Ear. B stage embryos were significantly higher as 3.9${\pm}$0.90 and 3.9${\pm}$0.90 with 2$^{nd}$ flushing time in total collected embryos and transferable embryos (p<0.05). Com M stage embryos were significantly higher as 3.7${\pm}$1.00 in 2$^{nd}$ flushing time and as 2.2${\pm}$0.76 in 3$^{rd}$ flushing time for recovered embryos (p<0.05). In transferable embryos, Com. M stage embryos were significantly higher (p<0.05) as 3.7${\pm}$1.00 in 2$^{nd}$ flushing time and as 2.2${\pm}$0.76 in 34$^{dr}$ flushing time, also. No. of degradation embryos was significantly higher as 2.2${\pm}$0.72 in 5${rd}$ flushing time, On the other hand, degradation embryos was not observed in transferable embryos (p<0.05). In conclusion, these results suggest that DEC method should effective methods for production of in vivo embryos using less flushing solution following perform until 4$^{rd}$ flushing time than conventional embryo collecting method. Also, it might be effectively collection of transferable embryos following more less procedure times compared to conventional embryo recovery methods.

• 한국수정란이식학회지
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• 제27권1호
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• pp.15-20
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• 2012

The objective of this study was to investigate the relationship between body weight, body condition score (BCS), blood urea nitrogen (BUN), glucose, cholesterol and number of transferable embryos for the purpose of improving reproductive performance in Hanwoo donors. Seventy five cows, at random stages of the estrous cycle, received a CIDR together with injection of 1mg estradiol benzoate and 50 mg progesterone, and gonadotropin treatment begann. Four days later, the animals were superovulated with a total of 28AU FSH (Antorin, 2AU = 1 ml) administered twice daily in constant doses over 4 days. On the 3rd administration of FSH, CIDR was withdrawn and 25 mg $PGF_2{\alpha}$ was administered. Cows were artificially inseminated twice after estrous detection at 12 hr intervals. The cows received $100{\mu}g$ GnRH at the time of 1st insemination. Embryos were recovered 7 days after the 1st insemination. In conclusion, cows with body weight < 400, 400~450 and > 450kg had number of transferable embryos of $4.2{\pm}1.7$, $6.1{\pm}2.7$ and $4.8{\pm}2.6$, cows with BCS <2.25, 2.25~2.75 and ${\geq}2.75$ had number of transferable embryos of $4.6{\pm}1.6$, $5.7{\pm}2.4$ and $5.1{\pm}2.7$ respectively. These data indicate that a body weight and BCS for superovulation of CIDR-treated Korean native cows does not affect the embryo yield.

• 한국수정란이식학회지
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• 제26권3호
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• pp.153-158
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• 2011

This study was performed in order to simplify the operation and minimize stress of donor and be readily available in the field with low cost and high quality embryos using the Direct Embryo Collection (DEC). Donors, at random stages of the estrous cycle, received a CIDR. 7 days later, 200 mg FSH was treated with 40, 30, 20, 10 mg FSH levels in declining doses twice daily by intramuscular injection for 4 days. On the 3rd day administration of FSH, 25 mg $PGF_2{\alpha}$ was administered and CIDR was withdrawn. After FSH injections were complete, donors were artificially inseminated twice at 12 hr intervals. The donor cattle received 250 ${\mu}g$ GnRH at time of 1 st insemination and embryos were recovered 8 days after the 1st insemination. Embryo collection from superovulated donors was performed to flushing by non-surgical methods of 3-way, 2-way and DEC (l-way). The average number of recovered embryos were 11.25${\pm}$0.63, 12.5${\pm}$0.65 and 11.75${\pm}$0.48 from operations of 3-way, 2-way and DEC methods, respectively. There were no significant differences among the embryo collection methods. Also, The average number of transferable embryos were 6.25${\pm}$0.48, 7.25${\pm}$0.48 and 7.25${\pm}$0.63 from each embryo collection procedures. The number of transferable embryos was no differences among the 3-way, 2-way and DEC methods, respectively. Meanwhile, the ratio of transferable embryos for all recovered embryos from DEC methods was higher as 61.7 % than 55.6 %, 58 % from methods of 3-way, 2-way. And the flushing solution required for recovering embryos by DEC method was significantly lower as 0.28${\pm}$0.32 1 than 1.8${\pm}$0.12 1, 1.75${\pm}$0.10 1 from 3-way, 2-way methods (p<0.05). Also, the time required for recovering embryos by DEC methods was significantly lower as 27${\pm}$2 min than 51${\pm}$3, 45${\pm}$2 min, respectively (p<0.05). In conclusion, these results suggest that DEC method for embryo collection may be effectively used for production of in vivo embryos using less flushing solution and, it might be effectively available in the field compared to conventional embryo recovery methods using 3-way or 2-way balloon catheter.

• 한국가축번식학회지
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• 제13권2호
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• pp.113-120
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• 1989

In order to determine the sex of preimplantation embryos prior to transfer in cattle, a series of experiments were carried out using 45 Holstein donor cows to examine the ovarian response on the gonadotropin and PGF2${\alpha}$, and the morphology of fresh embryos or frozen/thawed embryos after deep freezing at -196$^{\circ}C$. The sexing of embryos treated with the medium containing H-Y antiserum(10%, v/v) and FITC anti-mouse IgG(10%, v/v) were analysed by chromosomal analysis, and the sex of the embryos which survived were ascertain after delivering the pups. The results obtained were summarized as follows ; 1. The average number of developed follicle and corpus luteum per cow were 13.5 and 8.1, and the ovalation rate was 60.1%. 2. Of 220-ova recovered, 75(34.1%) were morula and 91(41.4%) were blastocyst, and the morphological normal and abnormal rate of ova recovered were 75.5% and 24.5%, respectively. 3. Of 39 frozen/thawed embryos, the scores of normal morula and blastocyst, after thawing were 79.2%(19/24) and 73.3%(11/15). The average rate of frozen/thawed embryos which appeared morphologically normal post thawing was 76.9%(30/39). 4. The sex ratio was measured using the embryos treated with immunofluorescence assay to examine the relationship between embryo developmental stage, sex ratio of morula stage embryo was 42.2%(19/45) fluorescing and 57.8%(26/45) non-fluorescing, on the other hand, the ratio switched to 46.8%(29/62) fluorescing and 53.2%(33/62) non-fluorescing embryo in blastocyst stage. The sex ratio was also measured between fresh and frozen/thawed embryos, fresh and frozen/thawed treated embryos were indicated 45.8%(38/83) fluorescing, 54.2%(45/83) non-fluorescing and 41.7%(10/24) fluorescing, 58.3%(14/24) non-fluorescing. This trend indicated the approximal sex ratio was 1 : 1. 5. The result of karyotype test showed the successful rate of sexing embryo is fluorescing and non-fluorescing was 21.2%(7/33) and 29.6%(8/27). The female to male ratio within 33 fluorescing was 28.6 : 71.4, and the ratio of 27 non-fluorescing embryos was 87.7 : 12.5. 6. Of the embryo transferred after assignment of H-Y phenotype, five of the fluorescing embryos survived to term, all was males. Whereas six non-fluorescing embryos also survived to term and the sexes of the calves were 1 male 5 female.

• Tuberculosis and Respiratory Diseases
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• 제45권4호
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• pp.723-735
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• 1998

배 경: 비타민 C가 동물실험에서 기도 과민성을 감소시키는 역할을 하는 것으로 보고되고 있는데 이는 비타민 C가 히스타민 분해를 촉진하고, cyclooxygenase 통로를 $PGF_{2\alpha}$에서 $PGE_2$로 이동시키며 평활근 수축을 감소시키고, 산화물의 환원제로 작용하는 사실에 기인한다고 한다. 또한 비흡연자에 비하여 흡연자에서 혈중 비타민 C 농도가 감소하는데 이는 비타민 C가 흡연시 생성되는 산화물의 환원제로 소모되며, 니코틴에 의해 분비되는 catecholamine, serotonin의 합성에 관여하며, 결핍된 식이로 인한 것으로 알려져있다. 연구자들은 기도 과민성이 있는 흡연자와 대조군인 비흡연지를 대상으로 비타민 C의 기도 과민반응에 대한 효과를 알아보고 비타민 C의 기도 과민성에 대한 역할을 알아보고자 본 연구를 시행하였다. 방 법: 17명의 흡연자와 8명의 비흡연자에서 기도 과민반응에 대한 비타민 C의 급성 효과를 알아보기 위하여 혈중 비타민 C 농도 측정을 위한 채혈과 폐기능 검사와 메타콜린 기관지 유발검사를 시행하고 비타민 C 3 g 투여 후 1시간 뒤에 상기 검사들을 다시 시행하였고 1주 후 기도 과민반응에 대한 비타민 C의 만성 효과를 알아보기 위하여 17명의 흡연자에서 1주 동안 매일 비타민 1 g 투여 후 혈중 비타민 C 농도 측정을 위한 채혈과 폐기능 검사와 메타콜린 기관지 유발검사를 시행하였다. 또한 기도 과민성에 대한 비타민 C 의 역할을 알아보기 위하여 l주 후 메타콜린에 반응하는 15명중 12명의 흡연자에서 비타민 C 3 g과 cyclooxygenase 억제제인 indomethacin 100 mg을 동시 투여 후 폐기능 검사와 메타콜린 기관지 유발검사를, 1 주 후 메타콜린에 반응하는 12명의 흡연자에서 indomethacin 100 mg만 단독 투여 후 폐기능 검사와 메타콜린 기관지 유발 검사를 시행하였다. 결 과: 전혈 비타민 C 농도를 측정한 결과 흡연자, 비흡연자 각각 $1.17{\pm}:0.22$ mg/dL, $1.14{\pm}:0.19$ mg/dL로 유의한 차이는 없었다(p>0.05). 흡연자 17명중 15명(88.2%)이 메타콜린 기관지 유발검사 양성이었고 이중 10명은 $PC_{20}FEV_1$이 8mg/mL 이하였고, 비흡연자군 8명중 7명(87.5%)이 기관지 유발검사 음성이었다. 기관지 유발집사 양성인 15명의 흡연자중 13명에서 비타민 C 3 g 복용 1 시간 후 기관지 반응이 유의하게 감소하였고 l일 비타민 C 1 g 씩 1 주간 복용 후에도 기관지 반응의 감소가 계속 유지되었다. 흡연자의 $PC_{20}FEV_1$과 전혈 비타민 C 농도 사이에는 유의한 상관관계는 없었다. 비타민 C 3 g 투여 후 흡연자에서 관찰할 수 있었던 기관지 반응의 유의한 감소가 indomethacin 100 mg 첨가 후 소실되었다(p>0.05). 결 론: 이상의 결과로 볼 때 혈중 비타민 C 농도는 흡연자군, 비흡연자군에 있어서 유의한 차이가 없지만 흡연자군은 비흡연자군에 비하여 기도 과민성이 유의하게 증가되어 있었으며, 비타민 C 투여에 의해 증가된 기도 과민성이 유의하게 감소하고 indomethacin 첨가 후 비타민 C의 이러한 효과가 소실됨으로 보아 비타민 C가 arachidonic acid 대사작용에 관여함으로 기도 과민성이 감소되리라 사료된다.

• 한국수정란이식학회지
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• 제27권4호
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• pp.229-235
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• 2012

This research was investigated the relationship, in high-producing Holstein donor cows, between the number of the transferable embryos and the blood serum concentrations of Blood Urea Nitrogen (BUN), glucose and cholesterol, which affect the nutritional state of cows. CIDRs were inserted into the vaginas of twenty two heads of Holstein cows, regardless of estrous cycle. Superovulation was induced using folliclar stimulating hormone (FSH). For artificial insemination, donor cows were injected with $PGF_{2{\alpha}}$ and estrus was checked about 48 hours after the injection. Then they were treated with 4 straws of semen 3 times, with 12-hour intervals. Embryos were collected by a non-surgical method 7 days after the first artificial insemination. The total numbers of ova collected from 3 experimental groups whose blood BUN concentrations were <10 mg/dl, 11~18 mg/dl and ${\geq}19$ mg/dl were 8.9, 12.5 and 19.0, respectively; whereas the numbers of transferable embryos were 5.8 + 1.9, 7.9 + 2.8 and 5.2 + 1.4, respectively. When glucose concentration was <60 mg/dl, the total number of collected ova was 9.9, which was smaller than when the concentration was 60~70 mg/dl or ${\geq}70$ mg/dl. When glucose concentration was 60~70 mg/dl, the number of transferable embryos was 7.1 + 2.4, which was slightly larger than the numbers 6.4 + 2.1 and 6.1 + 1.7 that were obtained when the concentrations were <60 mg/dl and ${\geq}70$ mg/dl, respectively ; however, the differences were not significant (p>0.05). When cholesterol concentrations were <150 mg/dl, 150~200 mg/dl and ${\geq}200$ mg/dl, the total numbers of collected ova were 11.2, 11.3 and 8.6, respectively. Whereas the numbers of transferable embryos were 7.1 + 2.1, 7.3 + 1.9 and 5.6 + 1.3, respectively ; however, the differences were again not significant (p>0.05). The result of this research showed no significant difference in ovum recovery rate and the number of transferable embryos according to major metabolite concentrations in high-producing Holstein donor cows. However, it is considered that the failure of maintaining proper nutritional status would cause the fall in in vivo embryo productivity.

• 한국가축번식학회지
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• 제23권1호
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• pp.13-18
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• 1999

정소내 성숙한 정자를 생산하는 전능성을 가진 정조세포는 체세포와 동일수준으로 외래 유전자를 삽입 가능한 것으로 알려져 있다. 그러나, 이들 세포가 반수체 이후의 단계로 분화한 경우에는 왜래 유전자를 삽입하기보다는 단순히 결합하는 능력이 있는 것으로 알려져 있다 . 따라서, 본 연구는 외래 유전자를 정소실질내 주입함으로써 형질전환 동물생산이 가능한지에 대하여 검토하기 위하여, 한쪽 정소를 거세한 한국 재래산 양을 사용하였다. Liposome /DNA 복합체를 1 : 2의 비율로 희석한 후 정소실실내에 주입하여 정자를 경유한 유전자 전이의 가능성을 확인하였다. 또한 동결보존한 정액을 인공수정하기 위하여 PGF$_2$$\alpha$(0.15mg/kg/BW)를 근육 주사함으로써 인위적으로 발정을 유도한 후, 인공수정을 이용하여 임신과 분만을 유도하였다. 이들 결과를 요약하면 다음과 같다. 1. PCR에 의하여, 유전자 도입 후 채취한 정액에서 외래유전자는 80일 이상 존재하였으며, 가장 높은 전이율은 40 일째 얻어졌다. 이들 결과는 정조세포에 외래 유전자가 성공적으로 삽입되었음을 제안하였다. 2. 23 두 (평균 96 % 의 발정 유기율)의 재래산양에게 인공수정을 실시한 결과 이들 중 4두가 임신되어 7두의 자양이 생산되었다. 3. 생산된 7두의 산자중 genome DNA를 추출하여 PCR 및 Southern blotting을 실시 한 결과 2두가 형질전환으로 확인되었다. 이상의 결과로서 정자를 매개로 한 정소실질내 외래 유전자의 주입법은 형질전환의 생산을 위한 매우 유용한 수단으로 사용 가능함을 제안하였다.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.59-59
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• 2003

재래산양은 우리나라 고유의 유전자원으로서 보존뿐만 아니라 개량이 절실히 요구됨에도 불구하고 첨단생명공학기술을 이용한 연구가 극히 미진한 실정이다. 본 연구는 재래산양에 있어서 과배란처리에 의한 미수정 난자의 회수율 개선과 최적의 quality를 확보하기 위하여 과배란 처리 후 회수시간이 난포의 발달과 난자의 회수율에 미치는 영향을 조사하였다. 공시동물은 체중 15kg 전후의 성숙한 미경산 재래산양으로서 발정동기화를 위하여 CIDR(Eazi Breed, InterAg, New Zealand)를 10일간 질내에 삽입하고 과배란 처리는 FSH를 CIDR 삽입 8, 9, 10일째에 12시간 간격으로 70mg을 감량법으로 투여하였으며, PGF2$\alpha$는 8일째에 FSH와 함께 투여하고 CIDR제거는 10일째에 제거와 동시에 hCG 400IU를 투여하였다. 수정란회수는 외과적인 방법으로 실시하였으며, 난관에서 관류방법으로 난자를 회수한 다음 난포에서 흡입방법으로 난포란을 회수하였다. hCG 투여 후 29~34시간과 46~50시간에 관찰된 난포수는 두당 10.29개(144/14) 및 7.83개(47/6)였다. 또한 난포로부터 난포란의 채란율은 각각 66.0%(95/144) 및 53.2%(25/47)였으며, 두당 6.79$\pm$0.9개 및 4.17$\pm$1.1개의 난자를 채란하였다. hCG 투여 후 회수시간에 따른 난관에서 회수한 난자 수는 29~34시간 및 46~50시간 후 채란시 배란점은 평균 4.36개(61/14) 및 7.33개 (44/6)였으며, 난관에서 난자 채란율은 39.3%(24/61) 및 29.5%(13/44)였으며, 재래산양 한마리당 평균 1.71$\pm$0.5개 및 2.17$\pm$1.4개의 난자를 채란하였다. 난관으로부터 회수한 난자의 회수율은 29~34시간째에는 61개의 배란점을 확인하였으나 회수한 난자수는 24개로서 39.3%의 회수율을 보였고 두당 회수율은 1.71$\pm$0.5개였으며, 46~50시간에서는 29.5%의 회수율과 2.17$\pm$1.4개의 두당 회수율을 보였다. 회수한 난포란의 등급에 있어서 29~34시간에 등급별 회수율이 각각 GI(27.4%), GII(14.7%), GIII(43.2%), GIV(14.7%)로 나타났으며, 46~50시간에는 GI(12.0%), GII(12.0%), GIII(28.0%), GIV(48.0%)로 나타났다. 이상의 결과로 볼 때 미성숙 난자의 회수는 hCG 투여 후 29~34시간이 적합한 것으로 생각된다.