• 한국발생생물학회:학술대회논문집
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• 한국발생생물학회 1998년도 제4차 학술발표대회 및 정기총회
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• pp.41-42
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• 1998

The changes in calcium binding protein(CBP) of mouse epididymal sperm during their post-testicular differentiation were analyzed by two-dimensional SDS-PAGE. According to dpididymal maturation, capacitation and acrosome reaction of spermatozoa, both quantitative and qualitative changes of CBPs in the epididymal sperm was detected. It suggested that the development of fertilizing ability of epididymal sperm was closely related to the changes in the CBPs profiles of sperm during epidiyaml transit.

• Clinical and Experimental Reproductive Medicine
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• 제26권3호
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• pp.447-456
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• 1999

Objective: The purpose of this study was to evaluate outcome of intracytoplasmic sperm injection (ICSI) using epididymal and testicular sperm in patients with azoospermia. Methods: From March, 1993 to May, 1999, a retrospective clinical analysis was done of a total of 140 cycles in 112 patients who underwent ICSI. Subjects were divided into three groups: ejaculated-ICSI group included 42 cycles in 34 patients with ejaculated sperm who underwent ICSI due to severe oligospermia and past history of failed or poor fertilization in the previous in vitro fertilization and embryo tranfer (IVF-ET) cycles, microsurgical epididymal sperm aspiration and intracytoplasmic sperm injection (MESA-ICSI) group included 50 cycles in 42 patients with congenital absence of the vas deferens (CAVD) or unreconstructable obstructive azoospermia and testicular sperm extraction and intracytoplasmic sperm injection (TESE-ICSI) group included 48 cycles in 36 patients with no spermatozoa which can be retrieved from epididymis or non-obstructive azoospermia. Results: Normal two-pronuclear fertilization rates were similar in three groups: 64.4% for ejaculated-ICSI group, 59.4% for MESA-ICSI group and 60.4% for TESE-ICSI group. The pregnancy rates were 26.2%, 26.0% and 25.0% respectively. There were no significant differences in the fertilization, cleavage, and clinical pregnancy rates among ICSI cycles using ejaculated, epididymal and testicular sperm. Conclusion: Epididymal and testicular sperm obtained in azoospermic patients can fertilize oocyte successfully and may lead to be similar fertilization rates and clinical pregnancy rates to ejaculated sperm.

• Molecules and Cells
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• 제28권5호
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• pp.441-446
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• 2009

During epididymal transit, mammalian sperm acquire selected proteins secreted by the epididymis. We previously showed that a disintegrin and metalloprotease (ADAM) 7 is expressed specifically in the epididymis and transferred to the sperm surface during epididymal transit. Here, we show that mouse ADAM7 secreted to the epididymal lumen is associated with membranous vesicles known as epididymosomes. Furthermore, we found that ADAM7 can be transferred directly from epididymal vesicles to sperm and that it is an integral plasma membrane protein in sperm. Thus, our study provides new information regarding the unique mode of secretion and interaction of ADAM7 during the epididymis-to-sperm transfer process.

• 한국가축번식학회지
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• 제21권3호
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• pp.247-253
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• 1997

본 연구는 돼지난포란의 체외수정시 정소상체 미부정자의 형태학적 정상정자 비율에 따른 수정율과 배발달율을 조사하고자 실시하였다. 그 결과는 다음과 같다. 1. 정소상체 미부정자의 형태학적 정상정자 비율을 10%, 10~30%, 50%로 구분하여 수정율과 배발달율을 조사하였던 바, 50%에서 각각 64%, 26%로 나타나 다른 두 군보다 현저히 높게 나타났다 ( 10%: 27%, 6%, 10~30%: 36%, 5%) (p<0.01). 2. 50% 이상의 정상정자를 가진 정소상체 미부정자를 100% (5$\times$105 cells/ml)로 조정하여 수정하였던 바, 정소상체 미부정자군의 수정율과 배발달율 (63%, 27%)은 동결 융해된 사출정자군 (56%, 35%)과 유사한 결과를 나타냈다. 3. 또한, 정소상체 미부정자군에서 난자와 정자의 비율 (1:6000, 1:6650, 1:7700, 1:10000)에 따라 수정률과 배발달율을 조사하였던 바, 난자당 정상정자 비율이 1:6000 (68%, 32%), 1:6650 (89%, 31%)일 때 가장 높은 수정율과 배발달율을 나타내었다. 따라서, 정소상체 미부정자의 형태학적 정상정자 비율이 50%일 때, 동결 융해된 사출정자와 유사한 배발달을 얻을 수 있으며, 이러한 결과로 미루어 볼 때, 정소상체 미부정자를 체외수정에 사용함에 있어서 정자의 형태학적 평가가 선행되어진다면 더욱더 효과적인 배발달을 유기할 수 있을 것으로 사료된다.

• 한국동물생명공학회지
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• 제37권2호
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• pp.130-135
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• 2022

Epididymal sperm cryopreservation provides a potential method for preserving genetic material from males of endangered species. This pilot study was conducted to develop a freezing method for tiger epididymal sperm. We evaluated post-thaw sperm condition using testes with intact epididymides obtained from a Siberian tiger (Panthera tigris altaica) after castration. The epididymis was chopped in Tyrode's albumin-lactate-pyruvate 1x and incubated at 5% CO₂, 95% air for 10 min. The Percoll separation density gradient method was used for selective recovery of motile spermatozoa after sperm collection using a cell strainer. The spermatozoa were diluted with modified Norwegian extender supplemented with 20 mM trehalose (extender 1) and subsequent extender 2 (extender 1 with 10% glycerol) and frozen using LN₂ vapor. After thawing at 37℃ for 25 s, Isolate^® solution was used for more effective recovery of live sperm. Sperm motility (computerized assisted sperm analysis, CASA), viability (SYBR-14 and Propidium Iodide) and acrosome integrity (Pisum sativum agglutinin with FITC) were evaluated. The motility of tiger epididymal spermatozoa was 40.1 ± 2.0%, and progressively motile sperm comprised 32.7 ± 2.3%. Viability was 56.3 ± 1.6% and acrosome integrity was 62.3 ± 4.4%. Cryopreservation of tiger epididymal sperm using a modified Norwegian extender and density gradient method could be effective to obtain functional spermatozoa for future assisted reproductive practices in endangered species.

• Asian-Australasian Journal of Animal Sciences
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• 제27권6호
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• pp.791-796
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• 2014

Cryopreservation of epididymal sperm is an effective technique to preserve genetic materials of domestic cats and wild felids when they unexpectedly die. However, this technique inevitably causes detrimental changes of cryopreserved-thawed spermatozoa, for example, by physical damage and excessive oxidative stress. L-carnitine is an antioxidant that has been used to improve sperm motility in humans and domestic animals. This study aimed to investigate the effects of L-carnitine on cat epididymal sperm quality following cryopreservation and thawing. After routine castration, cauda epididymides were collected from 60 cat testes. The epididymal spermatozoa from 3 cauda epididymides were pooled as 1 replicate. Spermatozoa samples (16 replicates) were examined for spermatozoa quality and then randomly divided into 4 groups: 0 mM L-carnitine (control), 12.5 mM, 25 mM and 50 mM L-carnitine. The sperm aliquots were then equilibrated and conventionally frozen. After thawing, sperm motility, plasma membrane integrity, DNA integrity and acrosome integrity were evaluated. The 25 mM L-carnitine significantly improved sperm motility compared with a control group (p<0.05), although this was not significantly different among other concentrations. In conclusion, supplementation of 25 mM L-carnitine in freezing extender improves cauda epididymal spermatozoa motility. The effects of L-carnitine on the levels of oxidative stress during freezing and thawing remains to be examined.

• 한국수정란이식학회지
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• 제33권4호
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• pp.321-326
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• 2018

In this study, two epididymal spermatozoa recovery methods in relation to sperm number, motility, viability and acrosome reaction were examined. Seven bulls were castrated and 7 testicles with epididymides were transferred to the laboratoy. Epididymis in each bull was randomly used for flushing and mincing methods with semen extender (Optixcell, IMV, France). The recovered spermatozoa with adjusted sperm concentration to $40{\times}10^6cells/mL$ was diluted with optixcell and cryopreserved. In experiment 1, the difference in the total number of spermatozoa using flushing and mincing methods was insignificant (2570.0 and $2505.2{\times}10^6cells/mL$, respectively). For experiment 2, the percentage of motile spermatozoa and motility parameters between flushing and mincing methods were studied through the use of sperm class analyzer after frozen-thawing. The percentage of total motile sperm between flushing and mincing methods was almost the same with $89.5{\pm}12.8$ and $91.4{\pm}7.9%$, respectively. The same is the case with experiment 3 wherein the viability and acrosomal integrity of frozen-thawed epididymal spermatozoa by flushing and mincing was insignificantly different. The results from the study showed that both flushing and mincing methods can be used for epididymal spermatozoa recovery in bull.

• 한국동물학회지
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• 제38권4호
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• pp.514-522
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• 1995

생쥐 정자의 수정능력 획득과정 및 첨체반응을 전후로 정자표면과 albumin의 상호작용을 조사하였다. 부정소 정자를 체외에서 배양하는 과정에서 정자표면에서 이탈된 단백질의 분석과 함께 FITC-bovine serum albumin으로 정자를 형광염색하여 정자표면에 대한 albumin의 결합양상의 변화를 조사하였다. 90분간 정자를 배양한 수 정자를 제거한 후 농축한 배양액내에 정자 또는 부정소액에서 기원한 여러종의 단백질가 함께 albumin이 다량으로 발견되었다 정자의 체외배양 과정에서 일어나는 albumin의 이탈은 배양액내의 $Ca^2$+과 무관하게 일어났다. BSA-FITC는 정소내 정자의 두부표면에 미약하게 결합한 반면 미부부정소 정자의 첨체표면에는 다량 결합하였다. $Ca^2$+-ionophore인 A23187으로 첨체반응을 유발한 정자의 두부 표면에서는 후첨체부위만이 강하게 염색되었다. 이러한 결과는 정소 및 웅성 생식수관니에서 정자표면에 부착된 albumin이 자성 생식수관을 거치는 동안 이탈됨을 시사하며 이러한 현상은 정자의 수정능력획득과 밀접한 관련이 있는 것으로 사료된다.

• Clinical and Experimental Reproductive Medicine
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• 제30권2호
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• pp.119-126
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• 2003

Objective: This study was carried out to compare the clinical outcomes of intracytoplasmic sperm injection (ICSI) in patients with obstructive azoospermia according to sperm retrieval site and technique; microsurgical epididymal sperm aspiration (MESA), percutaneous epididymal sperm aspiration (PESA), testicular sperm extraction by open biopsy (TESE). Methods: The outcomes of ICSI and IVF-ET were evaluated and compared among 3 groups. Seventy three men suffering from infertility due to obstructive azoospermia had 107 ICSI cycles using MESA (21 cycles in 15 patients), PESA (26 cycles in 17 patients) and TESE (60 cycles in 41 patients). Results: In the clinical outcomes in patients undergoing ICSI with epididymal or testicular sperm, there were no significant differences in fertilization rate (66.1% vs. 60.5%), cleavage rate (94.9% vs. 97.6%), cumulative embryo score (CES) (51.3 vs. 58.8), implantation rate (7.9% vs. 6.1), and clinical pregnancy rate per ET (30.4% (14/46) vs. 25.4% (15/59)) between both groups. Also, in the clinical outcomes in ICSI patients using MESA, PESA, TESE, there were no significant differences in fertilization rate (61.8%, 69.4%, 60.5%), cleavage rate (92.1%, 97.3%, 97.6%), CES (38.1, 52.0, 58.8), implantation rate (9.5%, 6.6%, 6.1%), and clinical pregnancy rate per ET (35% (7/20), 26.9% (7/26), 25.4% (15/59)) among 3 groups. Conclusion: When compared with MESA or TESE, PESA, the clinical outcomes were similar in ICSI patients with obstructive azoospermia whatever the origin or the technique of sperm retrieval. However, we considered PESA is more time-saving and cost effective for ICSI in patients with obstructive azoospermia.

• 한국가축번식학회지
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• 제21권3호
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• pp.239-246
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• 1997

본 연구는 돼지 난포란의 체외수정시 정소상체 미부정자의 형태학적 정상정자 비율에 따른 수정능의 차이를 조사하기 위하여 실시하였다. 그 결과는 다음과 같다. 1. 정소상체 미부로부터 회수된 전체의 정자중 형태학적 정상정자의 비율에 따른 ( 10%, 10~30%, 50%) 정자침입율과 다정자침입, 전핵형성율 그리고 난자에 침입한 평균 정자수를 조사하였던바, 50%의 정자침입율과 다정자침입은 82.4%와 87.4%로 10% (29.7%, 22.6%)와 10~30% (20.3%, 37.0%)보다 유의하게 높게 나타났다 (p<0.01). 또한, 공시된 난자의 전핵형성율도 50%이상의 형태학적 정상정자를 가진 실험군에서 유의하게 높게 나타났다(p<0.01). 2. 50% 이상의 정상정자를 가진 정소상체 미부정자를 100% (5$\times$105 cells/ml)로 조정하여 수정시킨 후 그 결과를 동결 융해된 사출정자와 비교하였던 바, 다정자침입과 전핵형성율은 정소상체 미부정자 (86.7%, 35.1%)와 동결 융해된 사출정자 (86.0%, 39.4%)간에 차이를 나타내지 않았으나, 정자침입율은 정소상체 미부정자가 79.7%로 동결 융해된 사출정자의 95.5%에 비해 유의하게 낮게 나타났다(p<0.01) 3. 또한, 정소상체 미부정자군에서 난자와 정자의 비율 (1:6000, 1:6650, 1:7700, 1:10000)에 따라 정자침입율과 다정자침입 그리고 전핵형성의 차이를 조사하였던 바, 수정시 난자당 정자의 수가 증가할수록 정자침입율, 다정자침입 그리고 난자에 침입한 평균 정자 수는 함께 증가하는 것으로 나타났다. 그러나, 전핵형성율은 난자당 정자의 수가 1:6000과 1:6650에서 높게 나타났다. 이상의 결과는, 돼지 난포란의 체외수정시 정소상체 미부정자를 사용했을 때, 50% 이상의 형태학적 정상정자를 가진 정자를 사용하면 동결 융해된 정자와 유사한 수정율을 얻을 수 있었으며, 정자의 형태학적 평가는 좀더 효율적인 수정능획득을 위해 선행되어져야 한다는 것을 시사한다고 하겠다.