• Clinical and Experimental Reproductive Medicine
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• v.12 no.2
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• pp.99-116
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• 1985

소의 번식효율을 증가시키기 위한 기초실험으로서 6%, 10% 및 20%의 bovine serum albumin 을 사용한 선인들의 방법과 tyrode액을 사용하여 시도한 저자의 방법으로 소의 원정액, 희석정액 및 일반시판 냉동정액으로부터 고활력정자를 분리수집하여 정자의 각종 성상과 광학현미경적 형태를 비교관찰한 결과는 다음과 같다. 1. 원정액으로부터 bovine serum albumin을 사용하여 분리한 정자는 대조군에 비하여 운동성, 운동성 정자수, 정상정자율 및 전진운동성이 현저히 높았고 정자회수율은 6%일 때 가장 높았다. 2. 원정액으로부터 bovine serum albumin을 사용하여 고활력정자를 분리한 후 냉동한 정액의 정자운동성, 정상정자율 및 전진운동성은 대조군에 비하여 현저히 높았고, 이러한 현상은 bovine serum albumin의 농도가 20%일 때에 가장 현저하게 나타났다. 3. Bovine serum albumin을 사용하여 분리한 고활력정자의 냉동전 및 냉동후의 광학현미경적인 기형율은 대조군에 비하여 현저히 낮았다. 4. Bovine serum albumin을 사용하여 분리한 고활력정자는 전자현미경으로 세포막의 확장 및 공포 형성, acrosome의 확장과 density loss의 변형율이 대조군에 비하여 낮았다. 5. 일반시판 냉동정액으로부터의 고활력정자의 분리는 bovine serum albumin을 사용할 때는 어려웠으나, tyrode액을 사용한 이 실험에서는 가능하였다. 6. 원정액, 희석정액 및 냉동정액의 고활력정자회수율은 tyrode 액을 이용하여 80분간 정치하였을때 현저히 높았다. 7. Tyrode 액을 이용하여 원정액, 희석정액 및 냉동정액으로부터 분리된 고활력정자의 운동성, 전진운동성 및 정상정자율은 대조군에 비하여 현저히 높았다. 8. Tyrode 액을 이용하여 원정액, 희석정액 및 냉동정액으로부터 분리한 고활력정자의 광학현미경적 기형율은 대조군에 비하여 현저히 낮았다.

• Journal of Embryo Transfer
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• v.11 no.2
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• pp.159-166
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• 1996

본 연구는 돼지 액상정액을 인공수정용 100ml 플라스틱 병에 보존하면서 BF5희석액과 Butschwiler 희석액 간에 보존 온도별 차이를 조사하고, BF5 희석액에서의 글리세롤 농도의 효과를 조사하여 돼지 액상정액을 좀더 장기간 사용할 수 있는 방법을 찾고자 실시하였다. 돼지 액상정액을 5$^{\circ}C$ 냉장고에 보존하면서 조사한 바에 의하면, 37$^{\circ}C$에서 0.5 및 2시간 배양후의 정자운동성은 전체 보존기간동안 BF5 희석액이 Butschwiler 희석액보다 유의하게 (P<0.05) 높게 나타났고, 정상첨체비율은 두 희석액간에 차이가 없었다. 돼지 액상정액을 15$^{\circ}C$에 보존하면서 조사한 바에 의하면, 3일부터 7일 보존시 까지 정자운동성과 정상첨체비율에 있어서 Butschwiler 희석액이 BF5 희석액보다 유의하데 높게 나타났다. BF5 희석액을 이용한 돼지 액상정액의 글리세롤 농도의 효과에 있어서는 최종 글리세롤 농도가 0, 2, 3, 및 5% 보다 1%일 때 가장 높은 정자운동성과 정상첨체비율을 나타내었다. 분만율, 복당 생존자돈수 그리고 출생시 평균 생시체중은 BF5 희석애과 Butschwiler 희석액간에 차이가 없었다. 이상의 연구 결과를 종합해 볼 때 BF5 희석액을 5$^{\circ}C$에서 Butschwiler 희석액은 15$^{\circ}C$에서 6-7일 동안 돼지 액상정액을 보존할 수 있었다.

• Korean Journal of Animal Reproduction
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• v.23 no.3
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• pp.191-203
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• 1999

The objectives of this study were to investigate 1) the effects of Selenium(Se), Vitamin E (Vit. E) or recombinant Bovine Somatotropin(rBST) administration on fresh and frozen/thawed semen characteristics and 2) the effect of taurine on frozen/thawed semen characteristics in Hanwoo sires Hanwoo sires were randomly assigned to five groups (1. control, 2. rBST, 0.09mg/kg body weight (BW), 3. Vito E 1,500IU/kg BW, 4. Se 0.l mg/kg BW, 5. Vit. E 1,500IU plus Se 0.1 mg/kg BW). The administration of Se, Vit. E and rBST for each experimental group were given 6 times at 15 days interval by intramuscular injection. The administration of Se, Vit. E or rBST in Hanwoo sires didn't affect semen volume and pH values, but sperm viability was significantly increased comparing to the control group. Also, frozen/thawed semen analysis showed that the sperm viability increased, but any other effects were not found in total sperm :lumber, motility and abnormality among treatments. The addition of taurine in semen freezing extender had a beneficial effects on frozen/thawecl semen characteristics in all groups. The administrations of rBST, Vit. E and Se did not affect the sperm capacitation and acrosome reaction, either the ratio of F pattern(uncapacitated and acrosome intact sperm) or AR pattern(capacitated and acrosome-reacted sperm), but the ratio of B patten(capacitated and acrosome intact sperm) of treatment groups was significantly higher than that of control group, These results indicated that the viability, motility and quality of semen in Hanwoo sires were slightly increased by the injection of rBST, Vit. E and Se, and the addition of taurine in semen freezing extender were also increased the semen characteristics after thawing.

• Journal of the Korean Society of Food Science and Nutrition
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• v.38 no.12
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• pp.1724-1731
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• 2009

This study was aimed to investigate the effect of phyto-extract fermented mixture (MP119) on the male sexual functions. The MP119 was evaluated for anti-impotency and anti-hypertensive effects via ACE (angiotensin converting enzyme) or PDE (phosphodiesterase) inhibition assay. $IC_{50}$ values of MP119 against ACE and PDE were 241.3${\pm}$35.5 ppm and 372.2${\pm}$33.8 ppm, respectively. To investigate the effect of testosterone expression by MP119, we performed cell media test using mouse Leydig-derived TM3 cells. Production of testosterone in TM3 cell was increased by MP119. Also, NO (nitric oxide) production of HUVEC (human umbilical vein endothelial cell) was increased when MP119 was added to the cultures. Forty male ICR mice were divided into 4 groups. MP119 was orally intubated for 7 days to group 1 and 3, and same volume of vehicle to group 2 and 4 as controls. After that, group 3 and 4 were intraperitoneally injected cadmium chloride at a single dose of 2 mg/kg. On the 8th experimental day, weights of testis, epididymis and seminal vesicle, number of sperm, concentrations of serum testosterone and cGMP were determined. The number of sperm, the concentrations of testosterone and cGMP were significantly increased in two experimental groups (group 1, 3). These results suggest that MP119 enhanced the sexual function of male mice, and could protect the sexual organs from the cadmium chloride as one of the endocrine disrupters.

• Reproductive and Developmental Biology
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• v.30 no.3
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• pp.219-224
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• 2006

The objective of this study was to investigate the toxic effects of Bisphenol A(BPA) and di-2 ethyhexyl phthalate (DEHP) as endocrine disrupters on sperm motility, viability, membrane integrity and abnormality during in vitro incubation of boar semen. Semen were randomly divide into 24 groups and healed with different concentrations of BPA md DEHP($1{\sim}100{\mu}M$) for 3, 6 and 9 hrs, respectively. The percentages of sperm motility and viability decreased by treatment time with both BPA and DEHP, and obiously differ from the controls. The percentages of sperm motility and viability significantly decreased by incubation with both $100{\mu}M$ of BPA and DEHP compared to control and other treatment groups(p<0.05). Sperm membrane integrity was significantly reduced by incubation with 10 and $100{\mu}M$ of BPA and DEHP, respectively(p<0.05), but sperm abnormality were not significantly affect both BPA and DEHP. These results indicate that high concentration of BPA and DEHP($>10{\mu}M$ can affect noxiously the sperm characteristics.

• Development and Reproduction
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• v.7 no.1
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• pp.9-13
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• 2003

A series of experiments were conducted to compare the effects of various diluents in cold storage on the sea urchin, Hemicentrotus pulcherimus sperm. Various diluents of glucose solutions, artificial sea water(ASW) and 50% ASW were used to store the sperm at 4$^{\circ}C$. The storage effect was evaluated using sperm activity index(SAI), survival rate of sperm and fertilization rate to egg. ASW and 1.2 M glucose were found to be better diluents which maintained high motility and survival rate of sperm f3r a storage period of 30 days. Optimal pH of diluent to store the sperm at 4$^{\circ}C$ is 7.0∼8.0. In order to keep high SAI and survival rate of sperm, addition of 400 ppm neomycin into the diluent revealed the best storage results.

• Journal of Embryo Transfer
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• v.22 no.2
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• pp.81-87
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• 2007

The purpose of the present study was to determine the preventive effects of the two antioxidant vitamin E and catechin on DEHP-induced disturbance of spermatogenesis in male rats. Rats at 4 weeks of age were randomly allocated into five groups with 20 animals per group. The first group was not any administrated as control. The second group was administrated DEHP (2 g/kg) daily for 14 days. The third group was administrated vitamin E (500 IU/kg) following DEHP treatment by the same method (daily for 14 days). The fourth group was administrated catechin (200 mg/kg) following DEHP treatment by the same method. The fifth group was co-administrated vitamin E (500 IU/kg) and catechin (200 mg/kg) following DEHP treatment by the same method. In order to determine the preventive effects, we examined pathological changes of testis with apoptotic index, and characteristics of sperm with computer assisted sperm analysis (CASA). Vitamin E and catechin supplementation were significantly prevented the testicular atrophy, apoptosis of germ cells in the seminiferous tubules and abnormal rate of sperm. Moreover, sperm concentration, viability and motility was significantly recovered in groups of alone and along with vitamin E and catechin. The results suggest that preventive effects of alone and along administration of vitamin E and catechin on DEHP-induced testicular atrophy damages have been demonstrated.

• Journal of Marine Life Science
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• v.8 no.2
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• pp.115-120
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• 2023

This study aims to find out a suitable extender and cryoprotective agent (CPA) for cryopreservation and its optimum concentration in order to conduct planned artificial seed production of Korean bullhead Pseudobagrus fulvidraco and to preserve superior sperm. Experiments were designed to investigate the effects of the different combinations of three extenders (I: 300 mM glycose, II: Kurokura extender, III: Li extender), four cryoprotectants (dimethyl sulfoxide, ethylene glycol, methanol and glycerol) and four concentrations (5, 10, 15, 20%) on the cryopreservation of Korean bullhead sperm. Postthawed sperm survival rate and sperm activity index (SAI) were detected to evaluate the effects of sperm cryopreservation. The optimal combination of extender and CPA for cryopreservation of Korean bullhead sperm was extender III + 10 and 15% methanol, resulting in a survival rate and SAI of 66.9 ± 8.7, 67.3 ± 13.1% and 2.6 ± 0.4, 2.6 ± 0.5 respectively, which was higher than had been achieved with other extenders and CPAs.

• Clinical and Experimental Reproductive Medicine
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• v.35 no.3
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• pp.203-212
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• 2008

Objective: During cryopreservation process, cold shock and cryo-injury affect the fertilizing capacity of the sperm by damaging cell membranes with loss of functional integrity. A longstanding concept for preventing the cryo-damage is to stabilize the plasma membrane by incorporating cholesterol. This study was to determine the effects of cholesterol in freezing media on the motility and functional integrity of human sperm after cryopreservation. Methods: Control group (non-cholesterol treated) and different concentrations of cholesterol-treated sperm (14 healthy males) were frozen and thawed. After freezing and thawing of sperm, the quality of sperm was evaluated by sperm analysis, acrosome reaction test and sperm chromatin structure assay. Results: When human sperm were incubated in sperm freezing medium (SFM) containing $0.5{\mu}g$ cholesterol and then freezing/thawing, the motility of sperm have significantly improved compared to those untreated cholesterol ($33.46{\pm}1.48%$ vs. $30.10{\pm}1.07%$, p<0.05). The rate of calcium ionophore-induced acrosome reactions in post-thawed sperm was significantly higher than that ($53.60{\pm}1.60%$ vs. $47.40{\pm}1.86%$, p<0.05) in SFM containing cholesterol. Sperm chromatin structure assay revealed that DNA damage to the sperm in the cholesterol-treated group was lower than that of non-treated group. Conclusion: These results suggest that increased cholesterol content of sperm plasma membrane by supplementation of cholesterol in SFM improves sperm motility, capacitation status, and DNA integrity. Therefore, addition of cholesterol into SFM could be a useful for protecting human sperm from cold shock and cryo-injury during cryopreservation.

• Korean Journal of Animal Reproduction
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• v.26 no.1
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• pp.1-7
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• 2002

The present study was carried out to investigate the effects of maturation media on penetrability of pig oocytes by liquid boar sperm coincubated with different sperm concentrations in a modified Tris­buffered medium (mTBM). Follicular oocytes collected from ovaries of prepubertal gilts were matured in a modified TCM-199 (mTCM-199) medium, modified Waymouth MB 752/1 (mWaymouth MB 752/1) medium or NCSU-23 medium. Oocytes (30~40) were transferred into each well of a Nunc 4-well multidish containing 0.5 $m\ell$ maturation medium. The sperm­ich portion of ejaculates with greater than 90% motile sperm were used in the experiment. The semen was cooled 22 to 24$^{\circ}C$ over 2 h period. The semen was diluted with Beltsville Thawing Solution (BTS) extender at room temperature to give 2$\times$10$^{8}$ sperm/$m\ell$ in 100 $m\ell$ plastic bottle. Liquid boar semen of 30 $m\ell$ in 100 $m\ell$ plastic bottle was kept at 17$^{\circ}C$ far 5 days. The sperm with greater than 70% motility after day 5 of storage were used for in-vitro fertilization (IVF). After 44 h maturation of immature oocytes in 5% $CO_2$in air at 38.5$^{\circ}C$, cumulus cells were removed and oocytes (30~40) were coincubated for 6 h in 0.5 $m\ell$ mTBM fertilization medium with five different (1$\times$10$^{6}$ , 2$\times$10$^{6}$ , 4$\times$10$^{6}$ , 6$\times$10$^{6}$, 10$\times$10$^{6}$ $m\ell$) sperm concentrations. At 6 h after IVF, oocytes were transferred into 0.5 $m\ell$ NCSU-23 culture medium fur further culture of 6 h. At 12 h after IVF, sperm penetration, polyspermy and male pronuclear formation of oocytes were evaluated. Oocytes of NCSU-23 maturation medium decreased polyspermy and increased male pronuclear formation compared to those of mTCM­199 and mWaymouth MB 752/1 maturation media. Of oocytes matured in NCSU-23 medium and inseminated in mTBM medium with 2~4$\times$10$^{6}$ $m\ell$ sperm concentrations, 50.8~50.9% showed sperm penetration, 13.3~19.5% polyspermy and 43.9~45.4% male pronuclear formation. In conclusion, we found out that oocytes matured in NCSU­23 medium and inseminated in mTBM medium showed superior in­vitro fertilization compared to those matured in mTCM­199 and mWaymouth MB 752/1 maturation media and inseminated in mTBM medium. The optimum sperm concentrations for in-vitro fertilization of oocytes matured in NCSU-23 medium by liquid boar semen stored at 17$^{\circ}C$ for 5 days were 2~4$\times$10$^{6}$ $m\ell$.