• 한국수정란이식학회지
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• 제25권3호
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• pp.149-154
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• 2010

The objective of this study was to investigate the effects of abnormal ovarian cycles after superovulation treatment of Hanwoo donors. Thirty six, at random stages of the estrous cycle, received a CIDR. 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 On the 3th administration of FSH, CIDR was withdrawn and 25 mg $PGF_2{\alpha}$ was administered.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 Ind insemination. Embryos were recovered 7 or 8 days after the 1st insemination. The cows were considered to have resumed ovarian cyclicity on the day of ovulation if followed by regular ovarian cycles. 50.0 percentage of the cows (18/36) had normal resumption of ovarian cyclicity (resumption within 40 days after superovulation), and 50.0% (18/36) had delayed resumption(resumption did not occur until>40 days after superovulation). Delayed resumption Type II (first ovulation did not occur until $\geq$ 40 days after superovulation, i.e. delayed first ovulation 33.3%) were the most common types of delayed resumptions. The mean numbers of total ova from < 10 and 10$\leq$ of corpora lutea (CL) was 7.3 and 13.9, respectively. The number of transferable embryos differed between < 10 and 10$\leq$ CL was 4.2 and 5.1, respectively. 11.1 percentage of the cows (4/36) did not resumption their ovarian cyclicity until 60 days after superovulation treatment.

• 대한의생명과학회지
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• 제4권1호
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• pp.27-33
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• 1998

세포내, 외 $Ca^{2+}$ 농도구배 유지에는 $Ca^{2+}$-ATPase와 $Ca^{2+}$-$Na^{+}$ exachangers가 주요한 기능을 한다고 알려져 있는데 특히 $Ca^{2+}$-ATPase의 기능에 대해 많은 연구가 행해지고 있다 $Ca^{2+}$-ATPase는 체세포에서 세포막에 위치하고 있으며 $Ca^{2+}$을 세포외부로 배출하는 기능을 함으로써 세포내부의 $Ca^{2+}$농도를 낮게 유지할 수 있도록 하는 기능을 담당하고 있다. 이러한 $Ca^{2+}$-ATPase는 포유동물의 정자에도 존재하고 있지만 그 기능에 대해서는 아직 많은 설명이 되어있지 않다. 본 연구에서 정자가 수정을 하기 위한 기능적인 능력이 $Ca^{2+}$ 농도와 관련된 변화와 얼마나 연관되어 있는가를 규명하고, 이러한 $Ca^{2+}$ 농도 조절이 원형질막의 중요인자인 $Ca^{2+}$-ATPase와는 어떠한 연관성이 있는가를 알기 위해 시도한 결과, $Ca^{2+}$-ATPase는 세포내, 외 $Ca^{2+}$의 농도구배를 조절함으로써 세포내 $Ca^{2+}$의 농도를 증가시켜 정자가 수정능 획득과정으로 빨리 전환하도록 유도하고 첨체 반응에 중요한 역할을 하는 것으로 판단되며, 세포외 $Ca^{2+}$ 농도가 높게 유지될 경우에도 정자의 첨체반응이 유도됨으로써 난자와 용이하게 수정을 할 수 있는 생리적 환경이 제공될 수 있다고 사료된다.

• 한국수정란이식학회지
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• 제17권2호
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• pp.129-136
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• 2002

본 연구는 체내, 체외 소 배반포기 배의 GMP vitrification 후 활력도의 비교와 한우 수정란을 몽골 소에 수정란이식 후 산자생산 가능성을 조사하고자 실시하였다. 한우 수정란은 체외수정란 또는 과배란처리에 의한 체내수정란을 생산하여 GMP vitrification 방법으로 동결 후 몽고로 수송하였다. 수란우는 CIDR과 $PGF_2\alpha$ 처리에 의하여 동기화를 유도하였다 체내수정란생산을 위하여 7두를 과배란처리하였다. 총 64개의 배반포기를 회수하였다. ($9.1\pm2.94$per cow). 체외수정란생산은 80.1% 분할율(174／217)과 40.8% 배반포기 발달율(71／174)을 얻었다. 체내수정란(93.7%; 45／48)의 동결융해 후 생존율은 체외수정란(82.5%; 52／63)보다 유의적으로 높았다(P＜0.05). 8두의 몽골 소에 2개의 수정란을 이식하여 5두가 이식 후 60일째 임신이 확인되었으나, 그 중 1두는 240째 유산을 확인하였다. 그 중 2두의 수란우에서 2두의 산자를 275일째 생산에 성공하였다. 이러한 결과는 GMP vitrification 방법은 체내, 체외수정란의 동결보존방법으로 이용될 수 있을 뿐만 아니라 동결융해란의 몽골 소에 이식 후 한우를 생산할 수 가능성을 확인하였다.

• Reproductive and Developmental Biology
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• 제35권3호
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• pp.335-339
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• 2011

The objective of this study was to investigate the relationship between concentration of urea nitrogen, glucose, cholesterol and number of transferable embryos for the purpose of improving reproductive performance in blood of Hanwoo donors. Fifty five, at random stages of the estrous cycle, received a CIDR. 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 3th 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 1nd insemination. Embryos were recovered 7 or 8 days after the 1st insemination. Cows with BUN < 10, 11~18 and ${\geq}$19 mg/dl had number of transferable embryos of $4.32{\pm}1.3$, $5.8{\pm}1.8$ and $4.7{\pm}2.1$ respectively. The mean numbers of total ova from < 10 and 10${\leq}$ of corpora lutea(CL) was 8.9 and 14.3, respectively. The number of transferable embryos differed between < 10${\leq}$ CL was 4.8 and 5.6, respectively.

• Reproductive and Developmental Biology
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• 제35권3호
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• pp.233-238
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• 2011

This study was performed to investigate the FSH levels for superovulation procedure in Korean Native Cattle (Hanwoo). The effectiveness of 200 mg and 400 mg of FSH to initiate superovulation was examined in Hanwoo. Donors, at random stages of the estrous cycle, received a CIDR 7 days later, 200 mg FSH group was treated with 40, 30, 20, 10 mg FSH levels in declining doses twice daily by intramuscular injection fur 4 days. Also, 400 mg FSH group was treated with 80, 60, 40, 20 mg FSH levels. On the 3rd day administration of FSH, 25 mg $PGF_2$ ${\alpha}$ was administered and CIDR was withdrawn. Donors were artificially inseminated twice at 12 hr intervals. The donor cattle received 250 ${\mu}g$ GnRH at time of 1st insemination and embryos were recovered 8 days after the 1st insemination. As a results, average number of CL treated with FSH 200 mg was higher as $20.9{\pm}1.20$ than $15.8{\pm}0.63$ for donors treated with FSH 400 mg, respectively(p<0.05). Treated group of 200 mg FSH level increased (p<0.05) the number of embryos recovered per procedure compared to 400 mg FSH level ($18.2{\pm}1.18$ vs. $12.38{\pm}0.52$, respectively). When treatment of 200 mg FSH was performed, average transferable embryos/ova increased (p<0.05) to $14.1{\pm}1.12$ from $6.8{\pm}0.33$ of treated of 400 mg FSH. Group of 200 mg FSH increased (p<0.05) to $8.3{\pm}0.76$ from $2.0{\pm}0.26$ in morula stage compare to 400 mg FSH group. Mean of total early blastocyst and expanded blastocyst stage embryos was similar (p<0.05) between the 200 mg and 400 mg FSH levels group ($4.7{\pm}1.19$ vs. $2.9{\pm}0.18$ and $1.2{\pm}0.40$ vs. $1.9{\pm}0.17$). These results suggest that 200 mg FSH level-based superovulation protocol with CIDR may be effectively used fur production of superior embryos in Hanwoo. In other words, the less level of FSH may be effectively applied for Hanwoo (Korean Native Cattle), because Hanwoo was smaller body size than beef or daily cow.

• Reproductive and Developmental Biology
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• 제37권1호
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• pp.35-40
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• 2013

The objective of this study was to investigate the effects of abnormal ovarian cycles after superovulation treatment of 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. The cows were considered to have resumed ovarian cyclicity on the day of ovulation if followed by regular ovarian cycles. Seventy two point seven percentage of the cows(16/22) had normal resumption of ovarian cyclicity(resumption within 40 days after superovulation), and 27.3%(6/22) had delayed resumption(resumption did not occur until>40 days after superovulation). Delayed resumption Type II(first ovulation did not occur until ${\geq}40$ days after superovulation, i.e. delayed first ovulation 13.6%) were the most common types of delayed resumptions. The mean numbers of total ova from < 10 and $10{\leq}$ of corpora lutea(CL) was $7.8{\pm}1.8$ and $12.7{\pm}2.7$, respectively. The number of transferable embryos differed between < 10 and $10{\leq}$ CL was $5.4{\pm}1.3$ and $8.1{\pm}3.4$, respectively. Four point five percentage of the cows(1/22) did not resumption their ovarian cyclicity until 60 days after superovulation treatment. 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.

• Journal of Nutrition and Health
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• 제35권1호
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• pp.5-14
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• 2002

Alteration in the syntesis or enhanced inactivation of nitric oxide(NO) can induce impairment of endothelial cell function. Insulin dependent diabetes mellitus(IDDM) is characterized by impaired endothelial function and vascular disease. NO is produced through L-arginine pathway To elucidate the hypothesis that the decreased production on NO in IDDM reflects vascular damage and the NO production can be manipulated by either dietary fat(7% of kg diet) or the oral supplementation with L-arginine(2g/kg bw), plasma markers for vascular endothelial damage and plasma lipid profiles were measured in streptozotocin(STZ)-induced diabetic rats. Diabetic or normal Sprague-Dawley rats were fed 6 different experimental diets for 4 weeks(SO : soybean oil, SOA: soybean oil + L-arginine supplementation, BT : beef tallow, BTA_ beef tallow + L-arginine supplementation, OV olive oil, OVA : olive oil + L-arginine supplementation). Plasma glucose, total cholesterel, HDL-cholesterol, LDL-cholesterol and triglyceride were measured. Endothelial markers, plasma von Willebrand factor(vWf), thromboxane B$_2$, and 6-keto PGF1$\alpha$ of aorta were measured by ELISA. Plasma NO production was evaluated through the measurement of nitrite by EIA. Feeding saturated fatty acid(SFA, BT) increased relative liver size(RLS) in diabetic rats compared to either polyunsatunted fatty acid(PUFA, SO) or monounsaturated fatty acid(MUFA, OV) The supplementation of L-arginine inhibited the liver and kidney enlargement in olive oil find diabetic rats. Plasma glucose was lower in diabetic animal find the olive oil compared to fed beef tallow and the supplementation L-arginine decreased it in diabetic rats find beef tallow significantly(p < 0.05). Plasma TXB$_2$ levels were increased due to diabetes and the value of beef tallow group showed highest value. Plasma vWf concentration of beef tallow group was higher value in normal rats and was elevated more in diabetes. In diabetic groups, the vWf concentration of olive oil group was lower than beef tallow or soybean oil group. The supplementation of L-arginine in diabetic rats decreased plasma TXB$_2$ and vWf levels significantly(p < 0.05). NO production was higher in normal olive oil fed rats and was tend to be decreased in diabetic rats and the supplementation of L-arginine recovered to normal value(p < 0.05), Olive oil supplemented with L-arginine tended to lower plasma total cholesterol and LDL-cholesterol after 4 week treatment. These results suggest that generalized vascular endothelial changes based on plasma TXB$_2$and vWf occurs in diabetic rats. and olive oil with L-arginine supplementation contributes to a better control of the hyperglycemia, endothelial changes and hypercholesterolemia accompanying diabetes as compared with beef tallow or soy bean oil in this rat model.

• 대한수의학회지
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• 제38권2호
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• pp.280-289
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• 1998

Voltage-sensitive ion channels contribute to establishment of the cell excitablity and the generation of the cellular function. At hamster oocytes in the primitive stage during developing process, an inward current elicited by voltage pulses was found to be carried mainly by $Ca^{2+}$. Even at present, $Ca^{2+}$ channels serve as the most probable route to pass this inward current but there is no evidence of the presence of this channels in eggs. To date, both the characteristic properties and the physiological role in the early stage of development remain unclear. Here we examined the characteristic properties of the inward current and changes in this currents at unfertilized oocytes, fertilized zygotes and two-cell embryos using whole-cell voltage clamp technique. The inward current carried reportedly by $Ca^{2+}$ was remained following removing external $Ca^{2+}$ but completely abolished by further replacement of impermeants such as tetramethylammonium ion ($TMA^+$) or $choline^+$ instead of $[Na^+]_0$. Tetrodotoxin did not affect on this inward current remained at $[Ca^{2+}]_0$-free condition. Removal of $Na^+$ ion out of the experimental solution clearly decreased the current. After adding 2mM $Ca^{2+}$ to the $Na^+$-free media, the inward current was restored. Interestingly, this current carried by either $Ca^{2+}$ or $Na^+$ was decreased by the reduction of intracellular $Cl^-$ concentration, or by $Cl^-$ channel blockers such as niflumic acid, DIDS and SITS. When $Cl^-$ concentration was lowered without changes in other ionic components, this inward current was reduced. At fertilized oocytes and two-cell embryos, the inward current carried by $Ca^{2+}$ and $Na^+$ was severely reduced. Also $Cl^-$ component could not be observed. From these results, the inward current is composed of $Ca^{2+}$, $Na^+$ and $Cl^-$ component, suggesting that the channel carrying this inward current is not selective specifically to $Ca^{2+}$. During early stage of development, the voltage-sensitive ion current seems not to contribute essentially to the cell cleavage and differentiation. The loss of $Cl^-$ component after fertilization suggests that $Cl^-$ may play a role in maintaining the viability of unfertilized ova.

• 농업과학연구
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• 제18권2호
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• pp.114-118
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• 1991

본(本) 연구(硏究)는 생체(生體) 및 실험실(實驗室)에서 성숙(成熟)된 돼지 난모세포(卵母細胞)의 체외수정(體外受精) 및 배양(培養)에 적합(適合)한 배지(培地)를 찾기 위하여 실시(實施)한 바, 그 얻어진 결과(結果)는 다음과 같다. 생체내(生體內)에서 성숙(成熟)된 난모세포(卵母細胞)에 대하여 10% FCS을 함유(含有)한 M199수정배지(受精培地)와 1% BSA를 함유(含有)한 TL Hepes 수정배지(受精培地)를 비교(比較)한 결과(結果), 수정율(受精率)은 TL Hepes 수정배지(受精培地)가 M199 수정배지(受精培地)보다 우수(優秀)하였으나, 다정자침입율(多精子侵入率)이 다소 좋지 않은 편이었다. 체외수정후(體外受精後) TL Hepes 세척(洗滌) 및 배양배지(培養培地)에서 48시간(時間) 배양(培養)한 결과(結果), 배양(培養)된 난자(卵子) 53개중(個中) 39개(個) (73.6%)가 분할(分割)되었으며, 분할(分割)된 수정란(受精卵) 39개중(個中) 31개(個) (79.5%)가 2~8세포기(細胞期)까지 균등(均等)하게 분할(分割)되었다. 미성숙난포란(未成熟卵胞卵)을 Waymouth 성숙배지(成熟培地)에서 배양(培養)했을 때가 TL Hepes성숙배지(成熟培地)에서 배양(培養)했을 때 보다 수정후(受精後) 더 많은 확장(擴張)된 정자두부(精子頭部)를 유도(誘導)할 수 있었으나, 대부분 다정자침입(多精子侵入)이 되었다. 48시간(時間) TL Hepes 세척(洗滌) 및 배양배지(培養培地)에서 배양(培養)한 결과(結果) 4세포기(細胞期)까지 발육(發育)을 시킬 수가 없었다.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 2001년도 추계학술대회 및 정기총회
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• pp.102-102
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• 2001

Taurine (2-ethaneaminosulfonic acid) is one of the major intracellular ${\beta}$ -amino acids in mammals and is required for a number of biological processes including membrane stabilization, osmoregulation, antioxidation, detoxification, modulation of calcium flux and neurornodulation. The taurine transporter (TAUT) which contains 12 hydrophobic membrane-spanning domains has been cloned from dog kidney, rat brain, mouse brain, human thyroid, placenta and retina. In this study, The TAUT cDNA from the human intestinal epithelial cell, HT-29 was cloned and sequenced. Reverse-transcription polymerase chain reaction (RT-PCR) was performed to amplify partial cDNA encoding human intestinal TAUT. The coding region of the PCR product was 732 bp long. The primers were designed to encode highly conserved amino acid sequences near the transmembrane domains III (IPYFIFLF) and Ⅵ (KYKYNSYR) both in human and mouse. The TAUT cDNA amplified was ligated into the pGEX 4T-1 expression vector. The resulting sequence of human intestinal TAUT cDNA (Accession number of NCBI Genebank is AF346763) was identical to the sequences of the TAUTs previously determined in the human placenta and retina except 3 base pairs from that of the reported human thyroid. TAUT specific antibodies were generated to use them as biological tools in the studies of the biological role of TAUT. Peptides of 149-162 amino acid residue (14 amino acids) of the TAUT were synthesized. The synthetic peptide used in this study was LFQSFQKELPWAHC. This region was chosen not only to avoid putative glycosylation sites but also to exclude regions of known homology with GABA transporters in the extracellular hydrophilic domains. The synthetic peptide, TAUT-1 was conjugated with carrier protein, kehole lympet hemocyanin (KLH) to use as an antigen. When used for immunization on a rabbit to produce polyclonal antiserum, the conjugates elicited high -titered specific anti-TAUT-1 antibodies, which reacted well with the ovalbumin (OVA) conjugated peptides in ELISA. The KLH-conjugated peptide was also used as immunizing antigen in BALB/c mice to produce TAUT specific monoclonal antibodies. From the culture supernatant of the hybridoma, the specificity of anti-TAUT-1 monoclonal antibodies was confirmed by ELISA. Further applications of more tools in TAUT expression analysis will be performed such as western blotting and flow cytometry.