• 제목/요약/키워드: SPERM

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Excretory-Secretory Products of Trichomonas vaginalis Cause Apoptosis in Mouse Sperm in Vitro

  • Keum, Jihyun;Roh, Jaesook;Ryu, Jae-Sook;Ryu, Ki-Young
    • Parasites, Hosts and Diseases
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    • 제60권5호
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    • pp.357-360
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    • 2022
  • Excretory-secretory products (ESP) of T. vaginalis have been shown to inhibit sperm motility, viability, and functional integrity, leading to a decreased fertilization rate in vitro. This study investigated whether T. vaginalis induce apoptosis and ultrastructural changes of sperm using flow cytometry and electron microscopy. Incubation of sperm with T. vaginalis ESP increased phosphatidylserine externalization and DNA fragmentation, and decreased mitochondrial membrane potential. Transmission electron microscopy of sperm incubated with ESP revealed abnormal features such as distorted heads, broken necks, and acrosomes exocytosis. This is the first report that demonstrates a direct impact of T. vaginalis ESP on sperm apoptosis and architecture in vitro.

난포액이 Sucrose 층을 이용한 정자의 Swim-up 분리에 미치는 효과 (Effect of Follicular Fluid on Sperm Swim-up Separation with Sucrose Layer)

  • 김경화;여영근;박영식
    • 한국수정란이식학회지
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    • 제13권3호
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    • pp.277-289
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    • 1998
  • 정자의 무분별한 이동을 충분히 억제하면서 운동성을 저해하지 않는 sucrose 층으로부터의 swim-up 분리체계를 구축하기 위하여, 정자의 이동에 장애가 될 수 있는 sucrose 층에 첨가되는 sucrose수준과 정자의 이동과 운동을 극대화시킬 수 있는 배양시간 및 sucrose 이중층의 형태를 조사하였던 바, 얻어진 결과는 다음과 같다. 1. 10mM의 sucrose 층은 정자의 swim-up 이동을 억제하였다. 2. Sucrose 층을 이용하지 않은 swim-up 분리에서 25분간 배양후 회수된 정자의 수가 유의하게 증가되었다. 한편, sucrose 층을 이용한 swim-up 분리에서 10분간 배양후 회수된 정자의 수가 유의하게 증가되었으며 정자의 운동성도 유지되었다 3. Sucrose 이중층 Type I과 Type II 간에 장애효과에는 유의한 차이는 없었으나, 장애공간이 짝은 Type II 이중층에서 시료 채취가 편리하였다. 정자의 주화적 이동과 관련하여 난포액의 기능 을 밝히기 위하여, 난포액의 희석과 열처리 및 난포액으로부터 추출한 단백질과 지질이 Type II Sucrose 층으로부터 10분간 배양한 정자의 swim-up 분리에 미치는 효과를 조사하였던 바 얻어진 결과는 다음과 같다 4. 난포액은 정자의 이동과 운동을 자극하였고 수정능획득정자를 유인하였으며, 특히 10% 난포액에서 수정능획득정자의 유인효과가 가장 크게 나타났다. 5. 저온 열처리 (56$^{\circ}C$, 30분)는 정자의 이동과 운동을 자극하는 난포액의 효과를 감소시키지 않았다. 6. 난포액으로부터 추출한 단백질 성분은 정자의 이동을 자극하였으나, 단백질 성분의 여과는 정자의 이동을 자극하는 단백질 성분의 효과를 감소시켰다. 7. 난포액으로부터 추출한 지질 성분은 정자의 이동과 운동을 유의하게 자극하지 않았다. 8. 난포액으로부터 추출된 단백질과 지질의 병용처리는 정자의 이동과 운동을 자극하는 난포액의 효과를 감소시켰다. 결론적으로 정자의 swim-up 분리에 sucrose 이중층을 이용할 수 있으며, 난포액은 sucrose 층으로부터 정자의 이동과 운동을 자극하고 수정능획득정자를 유인하였다. 특히 열내성을 가진 단백질 성분이 정자의 이동을 자극하였다.

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자주복 (Takifugu rubripes) 정자의 동결보존 (Cryopreservation of Tiger Puffer (Takifugu rubripes) Sperm)

  • 장윤정
    • 한국발생생물학회지:발생과생식
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    • 제1권1호
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    • pp.29-36
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    • 1997
  • Experiments were performed to study the effects of diluents, cryoprotectant, equilibration time, thawing temperature and addition of BSA and egg yolk. Among the various diluents, Alsever's solution was the best for sperm cryopreservation. A combination of Alsever's solution and 15% ethylene glycol showed the better results than others did. Sperm activity indection and survival rate gradually decreased with the equilibration time. The appropriate thawing temperature was 30 ${\pm}1^{\circ}$C. These results indicate that sperm cryopreservation methods can be developed in tiger puffer.

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Analysis of calcium binding proteins of mouse epididymal spermatozoa

  • Park, Seung-Ho;Gye, Myung-Chan
    • 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.

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돼지의 정액성상에 관한 연구 (Studies on Semen Quality of Boar)

  • 문승주;임계택
    • 한국가축번식학회지
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    • 제14권2호
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    • pp.141-146
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    • 1990
  • 12 crossbreed boars received 4 rations containing varying levels of lysine and DL-methionine. The results obtained from this study are summarized as follows ; 1. Semen volume, total sperm number of the treatment B, C and D were increased significantly (p<0.05) as compared with the treatment A but abnormal sperm percent of treatment B and C was decreased significantly (p<0.05) as compared with A. Sperm number and sperm mortility were not different from treatments. 2. Amino acids contents of sperm plasma were not different from treatments.

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精子 洗滌液의 pH와 洗滌頻度 및 수소個體가 精子洗滌液內 水素이온濃度와 소 精子의 尖帽反應에 미치는 影響 (Effects of pH of Washing Solution, Washing Frequency and Individual Bull on Proton Concentration in the Sperm Washed Solution and Sperm Acrosome Reaction)

  • 박영식;임경순
    • 한국가축번식학회지
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    • 제15권1호
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    • pp.7-13
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    • 1991
  • 본 實驗은 精子 洗滌液의 pH와 洗滌頻度 및 수소개체가 精子洗滌液內 水素이온과 精子尖帽反應에 미치는 影響을 究明하기 위하여 실시하였는바 그 결과는 다음과 같다. 1. 精子를 SHP液으로 4차례 反復洗滌하여 培養하였을 때, 洗滌液의 吸光度差間 變化는 2차 精子洗滌液에서 가장 높았다. 2. 精子를 pH 5.99, 6.38, 6.78, 7.10, 7.40, 7.69, 8.15, 8.45 및 8.83 SHP液으로 3번 洗滌하였을 때, 精子洗滌液의 吸光度差間 變化는 pH 7.69~8.83에서 유의하게 증가하였으며, 1차 精子洗滌液이 2차와 3차 精子洗滌液보다 유의하게 높았다. 3. Holstein, KNC 및 Hereford 精子를 pH 5.99, 6.38, 6.78, 7.10, 7.40, 7.69, 8.15, 8.45 및 8.83의 SHP液으로 3번 洗滌하였을 때, 精子洗滌液의 光度差間 變化는 Holstein이 KNC와 Hereford보다 유의하게 높았으며, Holstein은 pH 7.69~8.83에서, KNC과 Hereford는 8.15에서 각각 유의하게 증가하였다. 4. 精子를 pH가 6.8, 7.1 및 7.4인 SHP液으로 3번 洗滌한 다음 pH 7.4인 mTALP液에서 15분간 培養하였을 때, 精子洗滌液의 吸光度差間 變化는 1차 및 2차 精子洗滌에서 pH 7.1과 7.4가 6.8보다 유의하게 높았으며, 精子尖帽反應率은 pH 7.1 및 7.4가 각각 68.8 및 72.9%로 pH 6.8의 49.1%보다 유의하게 높았다(P<0.05).

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무력정자증 환자에서 인간 난포액과 SpermGrad를 이용한 정자처리법의 비교 (Comparison between Human Follicular Fluid and SpermGrad for Sperm Preparation in Asthenozoospermia)

  • 정연경;이정렬;문정희;김현준;한상훈;지병철;구승엽;서창석;김석현;최영민;김정구;문신용
    • Clinical and Experimental Reproductive Medicine
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    • 제33권1호
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    • pp.53-60
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    • 2006
  • 목 적: 보조생식술에서의 정자처리법으로서 인간 난포액을 이용한 swim-down 방법의 유용성을 확인해 보고자 하였다. 연구방법: 분당서울대학교병원 산부인과에서 불임평가 목적으로 정액검사를 시행할 때 정자무력증 (asthenozoospermia, sperm motility < 50%)을 보이는 12명의 남성을 대상으로 하였다. 이들에서 검사 후 남은 정액을 100% 인간 난포액을 이용한 swim-down법과 SpermGrad를 이용한 밀도차 분리법을 적용하여 각각 처리하고 컴퓨터 정액분석기를 이용하여 정액검사를 시행하였다. 결 과: 두 군 모두에서 운동성과 빠른 운동성 정자의 비율, VCL (curvilinear velocity), ALH (amplitude of lateral head displacement)치 및 과활동성 정자의 비율이 통계적으로 유의하게 상승하였고 LIN (mean linearity)치는 유의하게 감소하였다. 100% 인간 난포액을 이용한 swim-down법에서 정자의 운동성이 SpermGrad를 이용한 밀도차분리법에 비하여 유의하게 높게 나타났으며 ($81.2{\pm}4.7$ vs. $67.6{\pm}2.3$, p=0.02), 다른 변수들은 두 군에서 차이를 보이지 않았다. 결 론: 100% 인간 난포액을 이용한 swim-down법은 정자무력증을 보이는 경우 유용한 정자 처리법으로 사료된다.

산화스트레스에 노출된 정자의 생존성 및 운동성에 있어서 커큐민의 이중효과 (Dual effect of curcumin on viability and motility of bovine sperm exposed to oxidative stress)

  • 화정석;김은진;류지현;;박창윤;최창용;강다원
    • 한국수정란이식학회지
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    • 제31권3호
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    • pp.299-305
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    • 2016
  • Although cryopreservation of sperm is routinely used for clinical requirement, it has some problems, such as high generation of reactive oxygen species (ROS) and cold-shock. To reduce the detrimental damage in sperm, anti-oxidants were added to cryoprotectant for sperm. Curcumin is one of anti-oxidants, which are added in cryoprotectants. However, recent studies have demonstrated that curcumin decreases sperm viability and motility. This study was performed to identify the effect of curcumin on hydrogen peroxide ($H_2O_2$)-exposed bovine sperm, which were cryopreserved-thawed. In $H_2O_2$-exposed bovine sperm, reactive oxygen species (ROS) were significantly reduced by treatment with curcumin in a dose-dependent manner (p < 0.05). Among tested concentrations of curcumin (1 to $50{\mu}M$), 30 and $50{\mu}M$ curcumin showed anti-oxidant effect on $H_2O_2$-induced ROS generation. On the other hand, combination of 30 or $50{\mu}M$ curcumin with anti-oxidant $H_2O_2$ increased the percentage of apoptotic sperm compared to only $H_2O_2$ treatment. Sperm viability was also decreased in the combination of 30 or $50{\mu}M$ curcumin with $H_2O_2$ as judged by FDA/PI staining. $H_2O_2$-induced decrease in sperm progressive motility was recovered by treatment with $1{\mu}M$ curcumin. These results show that high concentration of curcumin has anti-oxidant effect, but it has also cytotoxic effect on bovine sperm. Sperm viability and motility might be more affected by cytotoxic signals of curcumin compared to antioxidant signals.

The Usefulness of Selected Physicochemical Indices, Cell Membrane Integrity and Sperm Chromatin Structure in Assessments of Boar Semen Sensitivity

  • Wysokinska, A.;Kondracki, S.;Iwanina, M.
    • Asian-Australasian Journal of Animal Sciences
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    • 제28권12호
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    • pp.1713-1720
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    • 2015
  • The present work describes experiments undertaken to evaluate the usefulness of selected physicochemical indices of semen, cell membrane integrity and sperm chromatin structure for the assessment of boar semen sensitivity to processes connected with pre-insemination procedures. The experiments were carried out on 30 boars: including 15 regarded as providers of sensitive semen and 15 regarded as providers of semen that is little sensitive to laboratory processing. The selection of boars for both groups was based on sperm morphology analyses, assuming secondary morphological change incidence in spermatozoa as the criterion. Two ejaculates were manually collected from each boar at an interval of 3 to 4 months. The following analyses were carried out for each ejaculate: sperm motility assessment, sperm pH measurement, sperm morphology assessment, sperm chromatin structure evaluation and cell membrane integrity assessment. The analyses were performed three times. Semen storage did not cause an increase in the incidence of secondary morphological changes in the group of boars considered to provide sperm of low sensitivity. On the other hand, with continued storage there was a marked increase in the incidence of spermatozoa with secondary morphological changes in the group of boars regarded as producing more sensitive semen. Ejaculates of group I boars evaluated directly after collection had an approximately 6% smaller share of spermatozoa with undamaged cell membranes than the ejaculates of boars in group II ($p{\leq}0.05$). In the process of time the percentage of spermatozoa with undamaged cell membranes decreased. The sperm of group I boars was characterised with a lower sperm motility than the semen of group II boars. After 1 hour of storing diluted semen, the sperm motility of boars producing highly sensitive semen was already 4% lower ($p{\leq}0.05$), and after 24 hours of storage it was 6.33% lower than that of the boars that produced semen with a low sensitivity. Factors that confirm the accuracy of insemination male selection can include a low rate of sperm motility decrease during the storage of diluted semen, low and contained incidence of secondary morphological changes in spermatozoa during semen storage and a high frequency of spermatozoa with undamaged cell membranes.

난자 세포질 내 정자 주입술시 부고환 및 고환 정자의 체외수정능력에 관한 비교 연구 (The Study on Comparison of Clinical Outcomes of Intracytoplasmic Sperm Injection in Patients with Epididymal Sperm and Testicular Sperm)

  • 성기청;강문주;김희선;오선경;구승엽;서창석;김석현;최영민;김정구;문신용
    • 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.