• Clinical and Experimental Reproductive Medicine
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• 제11권2호
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• pp.59-67
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• 1984

In vitro fertilization have been performed to know whether the frozen semen has fertilizing ability and can be used clinically. The results of cultured and developed embryos obtained are as follows: 1. The semen was frozen in three media for the good viability. The viability was more than 50% and the motility was also moderate (grade III), 2. As the 33 oocytes were collected from 45 follicles, the oocyte recovery rate was 73.3%. Among them, mature and immature ova were 5% each, and premature ova were 69.7%, When the first polar body was appeared, above ova were inseminated after adequate incubation with activated sperms. 3. The main components of three freezing medium containing egg yolk, glycerol and pyruvate respectively were the best for sperm viability, and Ham's F-10 medium was used for the fertilization and culture of eggs. 4. The results of in vitro fertilization of 33 ova, showed the second polar body developed in 12%, polyspermia in 24%, 1-cell embryo in 21% and 2-cell embryo in 9%. One mature ova developed to blastocyst via 16-cell to 32-cell embryo. The fertilization rate was 66%. 5. Above mentioned results represent that the frozen semen has fertilizing ability and can be used practically in the clinic.

• 한국가금학회지
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• 제27권3호
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• pp.215-220
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• 2000

This experiment was carried out to investigate the effect of reproductive ability on weeks and body weights of broiler breeder males. From 26 to 36 week, volume of semen and body weight were increased. Total concentration was highest at 36 week(P〈0.05), being 21.14$\times$10(sup)8 cells. Mean of total concentration was 14.66$\times$10(sup)8 cells and 11.59$\times$10(sup)8 cells in young(26∼55 weeks) and old(60∼89 weeks) broiler breeder males, respectively. Percentage of fertile eggs, viability and hatchability on weeks were 96.33, 89.55, 86.25% in the young broiler breeder males, 94.84, 91.73 and 86.97% the old broiler breeder males, respectively. The values were not significantly different between the young and old broiler breeder males. While the body weight of cocks was less than 3.5kg, it was not possible to collect semen. Semen collection rate was the best in 4.5∼5.0kg of males, total concentration was highest in 5.0∼5.5kg, volume of semen was highest in 5.5∼6.0kg body weight. More than 80% of the males were in the range of 4∼6kg body weight. Sperm motility was acceptable for breeding.

• Asian-Australasian Journal of Animal Sciences
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• 제10권6호
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• pp.609-613
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• 1997

The reproductive performance of forty two male broilers divided into three similar groups and fed on isocaloric and isonitrogneous diets containing 0, 10 or 20% water washed neem seed kernel meal (WWNSKM) was investigated from 20 to 32-wks of age. Results on semen characteristics revealed that feeding of WWNSKM led to significant (p < 0.05) reduction in semen volume, sperm concentration associated with increased incidences of morphological abnormalities in the spermatozoa when compared to that of the control birds. A drastic reduction in the fertilizing ability of spermatozoa was observed, the adverse effects being more at higher inclusion level of the cake. Hatchability of eggs also declined in the WWNSKM fed group. Histological examination of testes revealed a higher number of degenerating cells and poor spermatogenesis along with multinucleated giant cells in the seminiferous tubules of the testes of birds receiving the high dose of WWNSKM in diet. It may be concluded that the feeding of WWNSKM by incorporating in isocaloric and isonitrogneous diets to cockerels is associated with adverse effect on their fertility.

• Reproductive and Developmental Biology
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• 제36권3호
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• pp.147-154
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• 2012

본 연구에서는 제주흑우의 유전자원 보존과 증식에 있어 성공적인 인공수정을 위한 안정적인 동결정액 제조법을 수립하고 동결 융해 후 정자의 품질을 개선하기 위하여 제주흑우의 동결 정액 제조시 LDL, taurine, hypotaurine 그리고 trehalose를 첨가하여 이들이 동결 융해 후 정자의 성상에 미치는 영향에 대하여 알아보고자 수행하였다. 제주흑우의 정액 동결 시 LDL, LDL-taurine, LDL-hypotaurine 그리고 LDL-trehalose의 첨가는 ($63.40%{\pm}7.39$, $69.70%{\pm}6.12$, $67.25%{\pm}3.21$, $64.55%{\pm}2.43$) 대조구에($56.25%{\pm}6.42$) 비하여 모두 유의적인 생존율의 증가를 나타냈다 (p<0.05). 정자막 온전성 검사를 통한 꼬리막 팽창 정자의 비율은 LDL-taurine을 첨가 실험구에서 $70.55%{\pm}5.16$, LDL-hypotaurine 첨가 실험구에서 $64.45%{\pm}5.85$, LDL-trehalose 실험구에서 $64.50%{\pm}2.78$, LDL 단독 첨가 실험구에서 $61.65%{\pm}5.18$로 대조구 $51.90%{\pm}9.99$보다 모두 유의적으로 높은 결과를 나타냈다 (p<0.05). 동결 융해 후 정자의 첨체막 변화 양상에 있어서는 B pattern의 비율은 대조구와 실험구 모두 유의적 차이가 없었으며, F pattern은 LDL-taurine의 조합만이 대조구와 유의적인 차이를 나타내며 증가하였다 (p<0.05). 그러나 AR pattern의 비율은 LDL-taurine, LDL-trehalose, LDL-hypotaurine 그리고 LDL 처리의 순으로 대조구에 비하여 모두 유의적으로 낮은 수준의 AR pattern의 비율을 나타냈다(p<0.05). 정자의 수정능력 평가에 있어 LDL-taurine, LDL-hypotaurine 그리고 LDL-trehalose 모두 대조구에 대하여 유의적으로 높은 웅성전핵 형성율과 SFI를 나타냈으며 (p<0.05), 특히 LDL-taurine의 첨가는 LDL의 단독 처리에 비하여도 유의적으로 높은 수준의 웅성전핵 형성율과 SFI를 나타냈다 (p<0.05). 또한, decondenced sperm 비율에 있어 LDL, LDL-taurine, LDL-hypotaurine 그리고 LDL-trehalose 실험구 모두 대조구에 비하여 유의적으로 높은 결과를 나타냈다 (p<0.05). 본 연구의 결과는 제주흑우 정액의 동결정액 제조에 있어 안정적인 제조법을 공급할 수 있으며, 동결 융해 후 정자의 기능 개선 방법에 보다 많은 정보를 제공할 수 있을 것으로 사료된다.

• 한국발생생물학회지:발생과생식
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• 제1권1호
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• pp.57-65
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• 1997

생쥐의 부정소에서 진행되는 정자 성숙과정 동안의 첨체반응 능력의 변화를 조사하였다. 자발적 첨체반응 및 난포액, 프로게스테론, 또는 A23187에 의해 유발되는 첨체반응은 모두 정자의 성숙에 의존저긍로 일어났다. 두부 부정소의 매우 적은 정자만이 난포액 및 프로게스테론에 반응하여 첨체반응을 일으켰으며 체부 및 미부 부정소간에 첨체반응율에 차이가 없었다. 반면 A23187 처리시 상당수의 두부 부정소 정자가 첨체반응을 진행하였다. 이러한 결과에서 두부 부정소를 거쳐 체부 부정소에 도달한 생쥐 정자는 첨체반응 능력을 획득하여 이 과정에서 정자의 원형질막 표면에서는 첨체반응을 유발하는 물질과 상호작용에 필요한 변화가 진행되는 것으로 사료된다. 반면 정자내로의 $Ca^{2+}$ 유입 후 진행되는 막융합과 첨체내용물의 분비에 필요한 능력은 두부 부정소 정자도 일부 갖고 있는 것으로 사료된다.

• 한국가축번식학회지
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• 제9권2호
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• pp.148-152
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• 1985

When goat spermatozoa were preincubated for 4-6 h and 6 h in the uteri isolated from hamster and rat, and for 6 h in the hamster uterus in situ, they developed the ability to penetrate zona-free hamster eggs in vitro. Zona-free hamster eggs were not penetrated after insemination with goat spermatozoa preincubated in the isolated hamster uterus 4 h before and 2 h after expected time of ovulation, respectively. Zona-free hamster eggs were not penetrated after insemination with goat spermatozoa preincubated for 4 h in the isolated hamster uterus, but 10 and 18% of eggs were penetrated by spermatozoa preincubated for 5 and 6 h in the isolated uterus.

• 한국가축번식학회지
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• 제16권3호
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• pp.225-230
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• 1992

Many transgenic mice expressing human growth hormone gene were infertile. To investigate the infertility of these transfenic mice, it was looked into the estrus cycle and sexual behaviour and also tested through in vitro fertilization whether the germ cells of these mice normal or not. The infertile female transgenic mice were mated to the fertile males of ICR strain, but in almost all of them the vaginal plugs were not detected and their estrus cycles by vaginal smear were almost irregular which kept up estrus or diestrus stage. Many male transgenic mice did not have the ability of sexual behaviour. Therefore the viability of germ cells in infertile male transgenic mice was investigated by in vitro fertilization, but the sperm were normally fertilized with the eggs and the transgene of parent was passed on to the progeny. These results consequently suggest that the infertility of transgenic mice experssing human growth hormone gene may be due to the physiological activity of human growth hormone, not germ cells.

• Molecules and Cells
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• 제26권6호
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• pp.558-565
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• 2008

Although the fertilizing ability of spermatozoa is greatly reduced after freezing, complete understanding of alterations induced by cryopreservation has not been elucidated. The present study evaluates the effects of cryopreservation on the $Ca^{2+}$ handling of boar spermatozoa using several sperm activators. Intracellular $Ca^{2+}$ signals from single spermatozoa were measured using confocal $Ca^{2+}$ imaging of unfrozen samples and of other spermatozoa after having been frozen. Elevation of the external $K^{2+}$ concentration elicited a three times larger $Ca^{2+}$ increase in fresh spermatozoa than in cryopreserved spermatozoa. Caffeine elicited $Ca^{2+}$ transients with some oscillations in the fresh spermatozoa, but not in the thawed spermatozoa. Depletion of the $Ca^{2+}$ store with thapsigargin induced a rapid rise in $Ca^{2+}$ in the control but generated a smaller increase of $Ca^{2+}$ after thawing. Exposure to progesterone induced a biphasic rise of the $Ca^{2+}$ level in the fresh spermatozoa only. Sperm viability was reduced by cryopreservation. Resting $Ca^{2+}$ levels in fresh and cryopreserved spermatozoa were similar. Longer incubation (2.5 h) of thawed spermatozoa partly recovered the $Ca^{2+}$ response to the interventions. These results suggest that cryopreservation reduces the responsiveness of spermatozoa to depolarization, modulators of the internal $Ca^{2+}$ store and progesterone in terms of the $Ca^{2+}$ signal, thus providing a possible mechanism for reduced fertility observed in cryopreserved boar spermatozoa.

• Clinical and Experimental Reproductive Medicine
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• 제24권1호
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• pp.51-56
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• 1997

Recently, through the development of the primer extension preamplification(PEP) method which amplifies the whole genome, simultaneous multiple DNA analysis has become possible. Whole genome from each single cell can be amplified using 15 base oligonucleotide random primer. The greatest advantage of PEP-PCR is the ability to investigate several loci simultaneously and confirm results by analysing multiple aliquots for each locus. This technique led to the development of preimplantation genetic disease diagnosis using blastomere from early embryo, sperm, polar body and oocyte. In this study, we applied PEP-PCR in 20 cases of single amniocyte and 20 cases of single chorionic villus cell for the clinical application of the prenatal and preimplantational genetic diagnosis. We analysed 7 gene loci simultaneously which are 46, 47 exons related to dystrophin gene, two VNTR (variable number tandem repeat) markers using 5'dysIII, 3'CA related to dystrophin gene and DYZ1, DYZ3, DYS14 regions on chromosome Y. In all the tests, 97.5% of PEP-PCR amplifications with single cells were successful. We obtained 38/40 (95%) accuracy in gender determination through chromosome analysis comparison. Therefore, these results have significant implications for a sperm or oocyte analysis and prenatal or preimplantational genetic diagnosis.

• Clinical and Experimental Reproductive Medicine
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• 제24권1호
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• pp.67-81
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• 1997

To determine the concentration and the physiologic role of metal components in blood plasma and seminal plasma in relation to male infertility, the concentrations of twelve metal components in blood plasma and seminal plasma including Na, Mg, K, Ca, Cr, Mn, Fe, Cu, Zn, Se, Cd and Pb were measured by atomic absorbance spectrophotometery or ion selective electrode analysis. Semen and blood samples were obtained from a total of 110 men including 70 male infertility patients, 20 vasectomized persons and 20 fertility proven volunteers visited to the Male Infertility Clinic of Pusan National University Hospital. The concentrations of Ca, Zn, Mg, Cr and Cd in control group were higher in seminal plasma than in blood plasma, and additionally Pb were higher in infertility group. The concentrations of all metal components revealed no significant difference according to patients' age, resident, occupation, sperm density, motility and hormone level in blood plasma, but some metal components including Ca, Mg, Cu, Mn, Cd and Pb revealed a significant difference according to each these parameters except patient's age in seminal plasma. The concentrations of Mn, Cd and Pb in the vasectomy persons were higher than in the infertility group III including testicular and epididymal factors, but not in blood plasma. We conclude that the quantitative changes of metal components in the seminal plasma may have effects on not only spermatogenesis and sperm function, but also contribute to diagnostic parameter according to organ specificity of the metal in the male reproduction.