• Reproductive and Developmental Biology
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• 제35권1호
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• pp.85-91
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• 2011

The objective of this study was to evaluated the efficiency on sperm cryosurvival and ability of in vitro fertilization using Triladyl and Lactose Egg-Yolk(LEY) as extenders for cryopreservation of separated sperm by 65% percoll in miniature pig. Sperm viability was measured with SYBR-14/PI double stained sperm by flow cytometry. Ability on embryo cleavage rate and blastocyst development were observed by in vitro fertilization after frozen-thawing of sperm separated by 65% percoll. The experimental groups were designed that separated sperm by 65% percoll with Triladyl (ST) or LEY(SL) and unseparated sperm with Triladyl(UT) or LEY(UL) for cryopreservation. As a results, the viability was significantly(p<0.05) higher in ST(55.1%), SL(63.1%), UL(58.8%) than UT(38.2%) group. Sperm viability in SL(63.1%) group was significantly(p<0.05) higher than other experimental groups. On the other hand, embryo cleavage rate was significantly(p<0.05) higher in ST(79.1%), SL(83.2) than UT(74.1) and UL(75.7%) groups at 96h after in vitro fertilization. Blastocyst development was also significantly(p<0.05) higher in ST(21.5%), SL(20.9%) than UT(17.0%) and UL(18.8%) groups. In conclusion, cryopreservation of miniature boar sperm separated by 65% percoll were beneficial to viability and capacity on in vitro fertilization.

• 한국발생생물학회지:발생과생식
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• 제15권2호
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• pp.151-158
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• 2011

장기간 담수순화 사육한 감성돔 Acanthopagrus schlegelii(BFW) 정액의 화학적 특성과 염분 및 이온조성에 따른정자활성을 해수사육 감성돔(BSW) 정액과 비교하였다. BFW 정장의 화학적 특성은 대부분의 요인에서 BSW 정장과 차이를 보이지 않았으나, 삼투질농도는 각각 $307.0{\pm}4.6$, $337.3{\pm}10.1$ mOsm/kg으로 차이를 보였다. 염분에 따른 BFW 및BSW의 정자운동성는 0 psu에서는 운동성이 관찰되지 않았으나, 10 psu에서 낮은 운동성과 짧은 정자 운동지속시간을 보였다. 그러나 20, 32 psu에서는 높은 운동성과 긴 정자 운동지속시간 보였다. BFW와 BSW 정자의 SAI는 이온의 종류와는 상관없이 삼투질농도에 의존하여 변화하였으며, 인공해수와 비슷한 농도에서 높았다. 결론적으로 장기간 담수에서 사육한 감성돔의 정자의 운동개시요인은 환경수의 삼투질농도가 좌우하는 것으로 판단된다.

• 한국수정란이식학회지
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• 제28권2호
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• pp.87-93
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• 2013

The efficiency of artificial insemination (AI) for horses remains unsatisfactory. It is mainly because each process of AI causes a detrimental effect on semen quality. To sustain quality of semen properly, several factors including libido of stallions and sperm damage during sperm processing and preservation should be considered. Stallions with decent libido produce a high ratio of sperm to seminal plasma in their ejaculates, which is the ideal semen composition for maintaining sperm quality. Thus, to maximize the fertility rate upon AI, stallions should be appropriately managed to enhance their libido. Seminal plasma should have a positive effect on horse fertility in the case of natural breeding, whereas the effects of seminal plasma on both sperm viability and quality in the context of AI remain controversial. Centrifugation of semen is performed during semen processing to remove seminal plasma and to isolate fine quality sperm from semen. However, the centrifugation process can also result in sperm loss and damage. To solve this problem, several different centrifugation techniques such as Cushion Fluid along with dual and single Androcoll-E$^{TM}$ were developed to minimize loss of sperm and to damage at the bottom of the pellet. Most recently, a new technique without centrifugation was developed with the purpose of separating sperm from semen. AI techniques have been advanced to deliver sperm to optimal region of female reproductive tract at perfect timing. Recombinant equine luteinizing hormone (reLH) and low dose insemination techniques have been developed to maximize both fertility rate and the efficiency of AI. Horse breeders should consider that the entire AI procedure should be optimized for each stallion due to variation in individual horses for a uniformed AI protocol.

• Clinical and Experimental Reproductive Medicine
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• 제41권4호
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• pp.165-167
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• 2014

Objective: The objective of this study is to estimate the effects of Inclear, a feminine cleanser, on sperm motility. Methods: Semen samples were obtained from infertile male patients. Following liquefaction, the raw semen samples were diluted with Ham's F-10 nutrient mixture medium containing 0.4% human serum albumin solution at a ratio of 1:3. The semen samples were subsequently centrifuged to separate the seminal plasma from the serum. The supernatant was discarded, and the pellet was resuspended. The sample was again centrifuged to remove cell debris, and the supernatant was removed. The final pellet was gently loosened by resuspension and incubated in medium alone as a control, and in a 10% solution of the medium plus Inclear. A sampling time of 30 minutes was selected on the basis of sperm transport studies. Sperm motility was evaluated with computer-assisted sperm analysis. Results: A total of 20 samples were analyzed. The mean age of patients was $34.40{\pm}2.96years$. There was no difference in sperm concentration and motility in the two samples at 0 minute and 30 minutes of incubation. In both semen samples, the sperm concentration and motility decreased after an incubation period of 30 minutes. However, there was no statistical difference between the samples. Sperm concentration and motility were not significantly different between the control and Inclear samples after 0 minute and 30 minutes of incubation. Conclusion: Inclear has no negative effects on sperm motility. This product can be recommended to pregnancy planners for vaginal hygiene and as a vaginal lubricant.

• Clinical and Experimental Reproductive Medicine
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• 제40권1호
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• pp.23-28
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• 2013

Objective: The aim of the present study was to examine the relationship among male age, strict morphology, and sperm chromatin structure and condensation. Methods: Sperm samples from a total of 100 men underwent semen analysis, and sperm chromatin structure and condensation were assessed with toluidine blue (TB) and aniline blue (AB) tests. Results: Prevalence of strict morphology of less than 4%, and abnormal sperm chromatin structure and condensation did not show any statistically significant differences according to male age (p=0.605, p=0.235, and p=0.080). No significant correlation was demonstrated among age of male partners, strict morphology, and abnormal sperm chromatin structure using TB and AB tests. However, abnormal sperm chromatin condensation was positively associated with sperm chromatin structure (r=0.594, p=0.000) and showed negative correlation with strict morphology (r=-0.219, p=0.029). Conclusion: The tests for sperm chromatin condensation showed a significant association with strict morphology. Further study is needed to elucidate the relationship between clinical outcome and sperm chromatin tests.

• 한국패류학회지
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• 제27권4호
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• pp.377-386
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• 2011

The ultrastructures of germ cells during spermatogenesis and sperm morphology in male Mya arenaria oonogai, which was collected on the coastal waters of Samcheonpo, south coast of Korea, were investigated by transmission electron microscopic observations. In the early stage of the spermatid during spermiogenesis, a few granules and a proacrosomal granule, which is formed by the Golgi complex, appear on the spermatid nucleus, and then it becomes a proacrosomal vesicle. Consequently, it becomes an acrosome by way of the process of acrosome formation. The morphologies of the sperm nucleus type and the acrosome of this species have a curved cylindrical type and cone shape, respectively. The spermatozoon is approximately $48-50{\mu}m$ in length including a curved cylinderical sperm nucleus (about $2.65{\mu}m$ long), an acrosome (about $0.64{\mu}m$ in length) and tail flagellum ($40-45{\mu}m$ long). As some ultrastructural characteristics of the acrosomal vesicle, the peripheral parts of two basal rings show electron opaque part (region), while the apex part of the acrosome shows electron lucent part (region). These charateristics of the sperm belong to the family Myidae or some species of Veneridae in the subclass Heterodonta, unlike a characteristic of the subclass Pteriomorphia showing all part of the acrosome being composed of electron opaque part (region). Therefore, it is easy to distinguish the families or the subclasses by the acrosome structures. Exceptionally, In particular, a cylinder-like nucleus of the sperm is curved (the angle of the nucleus is about $20^{\circ}$), as seen in some species of Veneridae (range from $0^{\circ}-80^{\circ}$). The number of mitochondria in the midpiece of the sperm of this species are four, as one of common characteristics appeared in most species except for a few species in Veneridae in the subclass Heterodonta. Cross-sectioned axoneme of the sperm tail flagellum shows a 9+2 structure: the axoneme of the sperm tail flagellum consists of nine pairs of peripheral microtubules at the periphery and a pair of central doublets at the center.

• 대한한의학방제학회지
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• 제17권1호
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• pp.113-120
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• 2009

Objectives : This study focuses on the effect of the extract which We manufactured by adding Aliium sativum to Ojayeonjonghwan on the amount of semen, the number and the activity of sperm. Methods : We observed any changes in the amount of semen, the number and the activity of sperm after orally administering the test drug for thirty days and taking semen samples from a selected group of men - healthy men in their 20s, as well as men in their 40s who were feeling a decrease in energy. Results : This experiment showed the result that the amount of semen increased by adding Aliium sativum to Ojayeonjonghwan in the case of men in their 20s, but did not show a statistically significant result. It showed the result that the amount of semen increased by a statistically significant level as well as the activity of sperm. Consequently, the result indicated that orally administering the extract added Aliium sativum to Ojayeonjonghwan increased the amount of semen, the number and the activity of sperm for men in their 20s. Men in their 40s who are feeling a decline in energy generally decreased the amount of semen, the number and the ratio of activity of sperm. However the amount of semen remarkably increased and the number and the activity of sperm increased but the result was not statistically significant after orally administering the test drug for thirty days. Conclution : Eventually, the result after orally administering the test drug to the group of men in their 20s and 40s for thirty days indicated that both groups increased the amount of semen, number of sperm and activity of sperm to a statistically significant degree.

• Clinical and Experimental Reproductive Medicine
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• 제18권1호
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• pp.95-99
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• 1991

Swim-up and Wang's tube system are known methods of motile sperm selection and aerobic bacterial removal from the raw semen. This study was designed to evaluate the recovery rate of motile sperm, % normal morphology of sperm, the efficiency of bacterial removal after sperm preparation by the above two methods. The results were as follows. 1. There was more significant increase of sperm concentration in preparation by swim-up than Wang's tube (p<0.05). The concentration of sperm by swim up was changed from $82.5{\times}10^6/ml$ to $6836{\times}10^6/ml$, and Wang's tube was changed from $82.5{\times}10^6/ml$ to $36.0{\times}10^6/ml$. 2. There was significant increase in sperm motility after preparation by two methods in comparison with initial sperm motility (p<0.05), but no statistical difference between two methods was noted. The % motility of sperm by swim-up was increased from 66.1% to 95.7% and Wang's tube from 66.1% to 98.1%. 3. There was significant increase of % normal morphology of sperm in the samples prepared by two methods (p<0.05), from 49.2% to 85.3% in swim-up and from 49.2% to 92.1% in Wang's tube, but there was no statistical significance between them. 4. There was no bacterial growth in aerobic culture after preparation by two methods.

• Asian-Australasian Journal of Animal Sciences
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• 제14권2호
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• pp.155-162
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• 2001

Effect of the north (N) or south pole (S) of a magnet and dithiothreitol (DTT) or heparin sodium (H) on the orientation of bull sperm were studied. Sperm were collected from four Japanese Black bulls of Okinawa Prefectural Livestock Experimental Station were treated with various concentrations of DTT or H during the first and sixth days and were then exposed them to N or S with a magnetic field of 3,000 Gauss for 24 hours. Experimental results showed that both N and S significantly increased the perpendicular oriented percentage of bull sperm in most treatments not treated or treated with DTT or H. Anisotropy of magnetic field of bull sperm was the main cause which induced them to orient perpendicular to the magnetic direction. The results of the experiment also showed that, in most cases, the two polarities had the same effect on the orientation of bull sperm. However, in some cases, N exerted a stronger effect on the perpendicular percentage of bull sperm than S, such as with the 20 mM DTT treatment at the first and sixth days and the 100 units H treatment at the sixth day. These cases showed that the perpendicular oriented percentage of bull sperm at N was significantly higher than that of S. It was only the 100 units heparin treatment at S both at the first and sixth days that upward perpendicular orientation was higher than that at N. This might be due to the fact that protamin-DNA structure of a small number of bull sperm nuclei slightly changed because of heparin action.

• 한국가축번식학회지
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• 제26권4호
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• pp.361-368
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• 2002

The onset of pronucleus formation and DNA synthesis in porcine oocytes following the injection of porcine or murine sperm was determined in order to obtain insights into species-specific paternal factors that contribute to fertilization. After 44h in vitro maturation, spermatozoa was injected into the cytoplasm of oocytes. After injection, all oocytes were transferred to NCSU23 medium and cultured at 39'E under 5% CO2 in air. Similar frequencies of oocytes with female pronuclei were observed after injection with porcine sperm or with murine sperm. In contrast, male pronuclei formed 8 to 9 h following the injection of porcine sperm, and 6 to 8 h following the injection of murine sperm. After pronucleus formation maternally derived microtubules were assembled and appeared to move both male and female pronuclei to the oocyte center. A few porcine oocytes entered metaphase 22 h after the injection of murine sperm, but normal cell division was not observed. The mean time of onset of S-phase in male pronuclei was 9.7 h following porcine sperm injection and 7.4 h following mouse sperm injection. These results suggested that DNA synthesis was delayed in both pronuclei until the sperm chromatin fully decondensed, and the sperm nuclear decondensing activity and microtubule nucleation abilities of the male centrosome are cell cycle dependent.