• 한국임상수의학회지
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• 제7권2호
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• pp.497-500
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• 1990

This study was carried out determine whether Unopette can be used for the observation of sperm morphology and sperm Concentration. Rabbit sperm and frozen-thawed bovine sperm were observed with phase contrast microscope after dilution with Unopette acooriding to duration of preservation at 3～5$^{\circ}C$. Sperm using Unopette showed high normal sperm(％) than sperm using hematoxylin-eosin until 48 hours. Sperm using Unopette revealed no difference in sperm concentration until 24 hours, as compared with control sperm. As a result, Unopette was assessed as appropriate solution for preservation in terms of morphological observation and sperm concentration.

• 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.

• 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.

• 대한생식의학회:학술대회논문집
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• 대한불임학회 2001년도 제41차 추계학술대회
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• pp.57-62
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• 2001

One of recent advances of mammalian fertilization is the understanding of the molecular basis of fertilization. Several proteins localized in sperm nucleus or on sperm surface are necessary for the fertilization process. Protamines, sperm nuclear proteins, are required for normal sperm function that leads to fertilization. Fertilin and cyritestin are sperm surface proteins and essential for sperm-egg binding. Fertilin is also required for sperm transport in the female reproductive tracts. Metalloproteses on sperm plasma membrane are found to play a role in sperm-egg fusion. The functional analysis of these proteins provides a new insight into the molecular mechanisms underlying mammalian fertilization and male fertility.

• 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.

• 한국동물생명공학회지
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• 제38권1호
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• pp.38-46
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• 2023

In this study, an experiment was conducted in order to determine what cryopreservatives (CPVs) were more effective in supporting the motility and viability of sperm from experimental animals. The sperm of mice, rats, beagle dogs, and rabbits were frozen using different CPVs, including DMSO, TYB, and Sperm CryoProtec. The results from freezing the sperm of each laboratory animal in Sperm CryoProtec showed a high level of sperm motility and viability in sperm samples from mice, rats, and beagle dogs melted at the end of the first week. For rabbits, a high level of motility was observed in sperm thawed during the first week, whereas a high level of viability was observed in sperm thawed during the second week. The results of analysis of sperm motility and viability using different CPVs according to laboratory animals showed a significantly higher level of sperm motility (26.28%) and viability (36.20%) for mice in Sperm CryoProtec and the lowest levels of motility and viability were observed in DMSO (p < 0.05). Significantly higher levels of motility (27.94%) and viability (37.94%) were observed for rats in Sperm CryoProtec compared with TYB, which showed the lowest levels of motility and viability (p < 0.05). The study findings described above suggest that the selection of appropriate cryopreservatives is required for each experimental animal. This is because there are differences in the levels of sperm motility and viability of experimental animals depending on the CPVs that are typically used for freezing human sperm, including Sperm CryoProtec and TYB.

• Clinical and Experimental Reproductive Medicine
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• 제18권1호
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• pp.23-34
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• 1991

In order to study the sperm-egg interaction during fertilization process in mouse, the effects of sperm concentration, the duration of capacitation and insemination, the stages of maturation and development of eggs, and sperm extracts and BSA on sperm binding to egg were examined. Sperm-egg binding was increased depending on sperm concentration within the range of $10^3-10^6$ sperm/ml. It showed the most numbers of sperm-egg binding at 60min from the beginning of preincubation(capacitation) and insemination, respectively. During sperm capacitation, sperm-egg binding inhibitor was released from sperm into the incubation medium. Sperm extracts containing trypsin-like enzyme which is secreted through the acrosome reaction increased the binding. BSA in the culture medium showed a positive effect on the binding. It is suggested that physicochemical alterations of zona pellucida in the process of maturation and fertilization of eggs leaded to inhibition of sperm-egg binding.

• 한국수정란이식학회지
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• 제24권4호
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• pp.275-279
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• 2009

The beneficial effect of glycerol as a cryoprotectant, especially for sperm cryopreservation, has been shown in many studies. However, glycerol is toxic to living cells, and boar sperm in particular show greater sensitivity to glycerol than sperm from other domestic animals. Amides have been studied as alternative cryoprotectants for freezing stallion sperm. Sperm frozen in methylformamide or dimethylformamide as cryoprotectants show similar motility when thawed compared with sperm frozen in glycerol. We evaluated the cryoprotective effects of dimethylformamide on boar sperm freezing. To test the effect of amides, the concentration of boar semen was adjusted to $10^9sperm/mL$, and seminal plasma was removed using Hulsen solution. After centrifugation, the pellet was diluted in modified-Modena B extender. Lactose-egg yolk (LEY) extender was used as the cooling extender. The freezing extender was madeed aaddition of the optimal amount of glycerol and amides to LEY-Glycerol-Orvus ES Paste extender, and this extender was used for the second dilution. Diluted sperm were frozen in liquid nitrogen using the 0.5 mL straw method. Sperm frozen in extender with glycerol as a cderol were compared with those frozen in extender including the different amides. Sperm were tested for motility, viability, the sperm chromatin structure assay, and normal apical ridge after thawing. The percent of motile sperm diluted in glycerol was as high as that in the stallion study (61%). Dimethylformamide showed positive effects on sperm quality and was better than glycerol. Methylformamide provided similar sperm quality as glycerol. Therefore, dimethylformamide is useful for reducing cryoinjury in boar sperm and is expected to be useful as an alternative cryoprotectant.

• Asian-Australasian Journal of Animal Sciences
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• 제20권3호
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• pp.351-358
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• 2007

Carbohydrate-protein interactions are known to be important in gamete interactions. Several evidence indicated that a fucose-containing sulfated polysaccharide fucoidan was potential inhibitor of fertilization in vitro and thus fucose seemed to be part of the recognition signal of gamete interaction in mammals. In recent investigation we found that ${\alpha}$-L-fucosidase activity was present in boar spermatozoa and it was related to sperm binding to and penetration into zona pellucida (ZP) in vitro. The objective of this study was to determine the effects of monosaccharide L-fucose and polysaccharide fucoidan on sperm ${\alpha}$-L-fucosidase activity and relation to sperm-oocyte interaction in pig. Results indicated that the activity of sperm ${\alpha}$-L-fucosidase was largely inhibited (62%) when sperm suspension was treated with monosaccharide L-fucose. It also significantly inhibited the number of sperm binding to ZP (32%) and penetration into zona-intact oocytes (72%), but did not inhibit penetration into zona-free oocytes when fertilization medium contained L-fucose. The chlorotetracycline (CTC) assessment showed that L-fucose did not affect induction of sperm capacitation and acrosome reaction. In contrast, the activity of sperm ${\alpha}$-L-fucosidase was not inhibited when sperm suspension was treated with polysaccharide fucoidan but sperm-ZP binding was greatly inhibited (85%) and completely blocked sperm penetration into zona-intact or zona-free oocytes. The CTC assessment showed that fucoidan increased the F pattern and decreased the AR pattern sperm. These results suggested that the different inhibitory mechanisms were present between monosaccharide L-fucose and polysaccharide fucoidan on sperm-oocyte interaction, the inhibition effect of ${\alpha}$-L-fucose on sperm binding and penetrating into ZP caused sperm ${\alpha}$-L-fucosidase inhibited by ${\alpha}$-L-fucose.

• 한국수정란이식학회지
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• 제14권1호
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• pp.59-67
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• 1999

To make up the medium for quantitative selection of capacitated-sperm through sucrose layer, the effects of BSA, caffeine, heparin and progesterone on sperm swim-up migration and movement were examined. And the results obtained were as follows; 1. BSA of 4mg/ml in bMSS stimulated sperm migration and movement, and attracted capacitated-sperm. 2. Caffeine of 5mM in bMSS containing 4mg/ml BSA stimulated sperm movement and attracted capacitated-sperm. 3. Heparin of 20$\mu\textrm{g}$/ml in bMAA containing both 4mg/ml BSA and 5mM caffeine stimulated movement and capacitation of sperm. 4. Progesterone of 50$\mu\textrm{g}$/ml in bMSS containing all 4mg/ml BSA, 5mM caffeine and 20$\mu\textrm{g}$/ml heparin (BCHP-MSS) attracted capacitated-sperm. 5. Effect of BCHP-MSS on sperm on sperm attraction was not different from effect of 10% follicular fluid solution (FF-MSS) on sperm swim-up separation. In conclusion, bMSS with 4mg/ml BSA, 5mM caffeine, 20$\mu\textrm{g}$/ml heparin and 50$\mu\textrm{g}$/ml progesterone(BCHP-MSS) was a optimal condition for selection of capacitated-sperm through sucrose layer.