• 대한수의학회지
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• 제24권2호
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• pp.245-266
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• 1984

As a fundamental study to increase the reproductive efficiency in cattle, highly motile sperm were separated and collected from raw semen, extended semen and frozen semen by different methods using various concentrations of bovine serum albumin or Tyrode's solution. Various characteristics and light microscopic and electron-microscopic morphology of sperm separated by different methods were compared. The results obtained were as follows; 1. The sperm separated from raw semen using bovine serum albumin showed significantly high value in motility, motile sperm count, percent of normal sperm and progressive motility, as compared with control sperm and revealed the highest sperm recovery rate when separated with 6% bovine serum albumin. 2. The sperm motility, percent of normal sperm and progressive motility of the highly motile sperm frozen after being separated from raw semen with bovine serum albumin, showed significantly high value than those of a control sperm and especially found the highest value when separated with 20% bovine serum albumin. 3. Light-microscopic percent of abnormality was significantly low in the prefrozen and postfrozen highly motile sperm separated with bovine serum albumin, as compared with control sperm. 4. Electron-microscopic finding of the highly motile sperm separated with bovine serum albumin showed low percent of deformity in the dilatation and vesiculation of cell membrane, in dilatation and density loss of acrosome than in those of control sperm. 5. It was impossible to separate the highly motile sperm from frozen semen with bovine serum albumin, but it was possible with Tyrode's solution. 6. Recovery rate of highly motile sperm from raw semen extended semen and frozen semen was the highest when the sperm pellet stood in Tyrde's solution for 80 minutes. 7. The highly motile sperm separated from raw semen, extended semen and frozen semen with Tyrode's solution showed significantly high value in motility, progressive motility and percent of normal sperm, as compared with control sperm. 8. Highly motile sperm, when separated from raw semen, extended semen and frozen semen with Tyrode's solution, showed significantly low percent of microscopic abnormality as compared with control sperm.

• 대한수의학회지
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• 제37권4호
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• pp.925-934
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• 1997

This study was directed at inducing the production of antibodies by immunizing heifers with bovine sperm antigen and on measuring the serum antibodies using indirect immunofluorescence assay(IFA) and agglutination test. The effect of antisperm antibodies on fertilizing capacity of bovine spermatozoa was evaluated. 1. Three heifers between 12- and 15- month old were immunized with bovine spermatozoa or phosphate-buffered saline. In heifers immunized with bovine spermatozoa serum IgG level was highest between 3 weeks and 5 weeks postimmunization detected by IFA. The antibody levels persisted through week 7 and slowly declined until week 20 and then antisperm antibodies were localized on spermatozoa. The fluorescent antisperm antibodies were detected at 2~20 weeks and at 6~9 weeks postinoculation on acrosome and tail, respectively. Among 21 sera from repeat breeder cows, only one cow has shown positive antisperm antibody response detected by IFA. 2. In spite of vital rate of bovine sperm after swim-up was not significantly affected by different concentration of antisperm antibodies in sera, the numbers of bovine sperm after swim-up were significantly reduced in proportion to the increased concentration of antibodies. Above 1/512 dilution of antibody neither influence on vital rate and numbers of bovine sperm nor sperm agglutination after swim-up. The study has also shown that the vital rate and number of sperm after swim-up and capacitation were also significantly reduced by the addition of antisperm antibodies. Although antisperm antibodies did not influence on the acrosome reaction rate of sperm during swim-up, did significantly reduce the sperm acrosome reaction rate after capacitation. The studies have resulted that the bovine antisperm antibodies can prevent the sperm motility by agglutination and block the capacitation and acrosome reaction of bovine sperm.

• 한국가축번식학회지
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• 제14권4호
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• pp.303-308
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• 1990

To investigate the effect of heparin on sperm capacitation. The acrosome reaction of bovine sperm which were incubated in mTALP containing heparin and the in vitro development of bovine follicular oocytes which were cocultured with heparin-treated sperm were evaluated, and the resutls were as follows : 1. When bovine fresh sperm were incubated in mTALP solution containing 0, 5, 10 and 25$\mu\textrm{g}$/ml heparin for 15 to 840 minutes, there was no significant difference between motilies of heparin-treated sperm and untreated sperm, but the acrosome-reaction rate of heparin-treated sperm was significantly higher than that of untreated sperm. Moreover the acrosome reaction rate of was sperm treated with heparin was significantly increased after incubating for 15 minutes. 2. When fresh sperm were incubated in mTALP solution containing 10$\mu\textrm{g}$/ml heparn for 840 minutes, the motility of sper incubated for 840 minutes was lower than those of sperm incubated for 0, 15, 60 and 120 minutes, but the acrosome-reaction rate of sperm incubated for 840 minutes was higher than those of the others. 3. When frozen-sperm were incubated in mTALP solution containing 10$\mu\textrm{g}$/ml heparin for 120 minutes, the acrosome reaction rate of sperm was significantly increased after incubating for 15 minutes. 4. When fresh sperm treated with heparin were cocultured with bovine follicular oocytes, 16.7 to 23.7% of the oocytes were developed to 2-8 cell stage.

• 농업과학연구
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• 제14권2호
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• pp.413-421
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• 1987

토끼에서 bovine serum albumin gradients로 성(性)의 전선택(前選擇)이 가능(可能)한지를 알아보기 위하여 6%, 10% 및 20%의 농도(濃度)를 달리한 bovine serum albumin을 사용한 방법(方法)으로 고활력정자(高活力精子)를 분리수집하여 정자(精子)의 성상(性狀)과 분리된 정액(精液)의 인공수정시(人工授精時)에 있어서 자토(仔兎)의 성비(性比)를 비교관찰한 결과는 다음과 같다. 1. Bovine serum albumin을 사용하여 분리(分離)된 정자(精子)는 임신율(妊娠率)에 있어서는 대조군에 비해 높은 성적(成績)을 나타내었으나, 자토(仔兎)의 성비(性比)에 있어서는 변화를 일으키지 못하였다. 2. Bovine serum albumin에 의해 분리(分離)된 정자(精子)는 대조군에 비하여 운동성(運動性), 정상정자율(正常精子率) 및 전진운동성(前進運動性)이 현저히 높았으며, 정자회수율(精子回數率)은 6%일 때 가장 높았다. 3. 원정액(原精液)으로부터 bovine serum albumin을 사용하여 고활력정자(高活力精子)를 분리(分離)한 후(後) 냉동(冷凍)한 정액(精液)의 융해후(融解後) 정자운동성(精子運動性), 정상정자율(正常精子率) 및 전진운동성(前進運動性)은 대조군에 비하여 현저히 높았다.

• Journal of Animal Science and Technology
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• 제57권4호
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• pp.16.1-16.7
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• 2015

A well-characterized sperm specific protein of the Member of immunoglobulin superfamily, IZUMO1, has crucial role in fertilization by mediating sperm binding to the egg plasma membrane in the mouse. However little is known about IZUMO1 in bovine. Here, we describe the molecular cloning and expression analysis of bovine IZUMO1 (bIZUMO1). RT-PCR and Western blot analysis of the bovine tissues indicated that bIZUMO1 was specifically expressed in the testis and sperm, Furthermore, the result of our biotinylation assay from ejaculated bovine sperm strongly suggest the assumption that bIZUMO1 is localized on the cell surface. These data imply the potential role of bovine IZUMO1 in mammalian fertilization.

• 대한수의학회지
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• 제29권1호
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• pp.57-60
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• 1989

This study was carried out to evaluate the possibility of sex preselection by gradients methods using bovine serum albumin in rabbits. Artificial insemination was performed with sperm from the top and bottom layer of rabbit semen separated by bovine serum albumin gradients. Various characteristics of separated sperm, and the conception rate and secondary sex ratio at artificial insemination with separated sperm were compared. The results obtained were as follows. 1. The sperm from the bottom layer showed significanty high value in motility, percent of normal sperm and progressive motility as compared with control sperm and the sperm and the sperm from the top layer. 2. The conception rate of sperm from the bottom layer was higher than that of the top layer. But secondary sex ratio was not altered by this methods.

• 한국수정란이식학회지
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• 제14권1호
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• pp.23-29
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• 1999

The objectives of the study were to establish sperm separation method and duration of insemination for bovine IVF. Oocytes from slaughterhouse ovaries were matured and fertilized using general protocol. After 18 or 42 h of insemination, six to ten embryos were placed into a 30${mu}ell$ drop of each medium, and the embryos were examined 7~10d post in semination without medium renewal. First, we compared Percoll gradient will swim-up technique for sperm separation. There was no difference in cleavage rates between them, but the development rates over morula stage of oocytes fertilized with sperm separated by Percoll gradient was significantly higher than that sperm selected by swim-up technique (p<0.05). Second, we evaluated development of bovine embryos derived from the IVF procedure with different durations(18 vs 42 h) of fertilization. There was also no difference in cleavage rates, but the development to blastocyst stage of oocytes exposed in cleavage rates, but the development to blastocyst stage of oocytes exposed to sperm for 42 h was significantly higher than that exposed for 18 h (p<0.05). In conclusion, Percoll gradient can be used for sperm selecton, improving of embryonic development. Also, 42h of IVF may improve the development of bovine embryos.

• 한국가축번식학회지
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• 제22권3호
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• pp.213-219
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• 1998

This study was carried out to investigate on the improvement of fertilizing ability of in vitro matured oocytes from sperm density, motility and polyvinylpyrrolidine (PVP) concentration, by intracytoplasmic sperm injection(ICSI) into the bovine oocytes. 1. The in vitro fertilization and cleavage rates of oocytes from 1.0, 2.0, 3.0, 5.0 ($\times$106/ml) sperm concentration by IVF and ICSI of bovine oocytes were 45.0%~65.0%, 65.0%~90.0% and 10.0%~30.0%, 35.0%~70.0%, respectively. 2. The in vitro fertilization and cleavage rates of oocytes from 20, 40, 60, 80% of sperm motility by IVF and ICSI of bovine oocytes were 47.8%~75.0%, 78.3%~90.0% and 8.7%~25.0%, 34.8%~70.0%, respectively. 3. The in vitro fertilization and cleavage rates of oocytes from 0.01, 0.02, 0.03, 0.05% of PVP concentration by microinjection of single into the bovine oocytes were 72.7%, 90.9%, 83.3%, 76.9% and 45.5%, 72.7%, 58.3%, 61.5%, respectively and these values of 0.02% addition of PVP were higher than other concentrations of PVP. 4. The in vitro fertilization and developmental rates of oocytes by IVF and ICSI methods were 63.3%~64.6%, 26.7%~29.2% and 88.2%, 47.1%, respectively. This ICSI method was improved high fertilization rates of bovined oocytes.

• 대한수의학회지
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• 제39권1호
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• pp.220-225
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• 1999

The aim of these experiments was to investigate the effects of bovine oviduct epithelial cells (OEC) derived from different segments to bind sperm binding and maintain their motility in vitro. In experiment 1, the number of sperm attached to OEC derived from isthmus or ampulla, the motility of unattached sperm during co-culture and fertilizing ability were assessed. In experiment 2, heparin treated sperm (hsp) or no treated sperm (nsp) were used to evaluate OEC binding ability of capacitated sperm. In experiment 1, regardless of their origin, approximately 65% of the sperm were attached to OEC within 2h. From 6h of co-culture, the numbers of unattached sperm on ampullary OEC were significantly higher than those on isthmic OEC (p<0.005). From 12h of co-culture, the motility of unattached sperm on isthmic OEC were significantly higher than those on ampullary OEC(p<0.05). The cleavage rate of oocytes inseminated on OEC derived from isthmic segment was also significantly higher than those from ampullary segment (p<0.01). In experiment 2, the numbers of unattached hsp on OEC were significantly higher than those of controls (p<0.01), between 2-24h examination. From 12h of co-culture, the motility of unattached nsp were significantly greater than those of hsp (p<0.01). These results show that bovine OEC derived from the isthmus play more important role(s) for sperm binding, maintaining motility and fertilization in vitro than those from the ampulla, and heparin induced capacitation may change sperm binding ability on OEC in vitro.

• Clinical and Experimental Reproductive Medicine
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• 제11권2호
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• pp.69-76
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• 1984

In order to obtain high proportion of Y sperm the semen was laid over 6%,10% and 20% bovine serum albumin. Separated sperm were stained with quinacrine mustard in order to see F - body which could be seen in human Y sperm. But we could'nt find F-body in the bull sperm. So sperm were compared with size of sperm head. As a result of observation separated sperm was small in size of length and width of sperm head as compared with control sperm. So it was found that the proportion of Y sperm showed a marked increase in separated layer. Then the higher albumin density was, the higher the proportion of Y sperm which had smaller head and faster motility was. But the higher albumin density was, the lower the recovery rate of sperm was. So it was hard to separate Y sperm in oligospermia.