• Asian-Australasian Journal of Animal Sciences
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• 제26권2호
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• pp.178-188
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• 2013

The glycosaminoglycans (GAGs) present in the female reproductive tract promote sperm capacitation. When bovine sperm were exposed to 10 ${\mu}g/ml$ of one of four GAGs (Chondroitin sulfate, CS; Dermatan sulfate, DS; Hyaluronic acid, HA; Heparin, HP) for 5 h, the total motility (TM), straight-line velocity (VSL), and curvilinear velocity (VCL) were higher in the HP- or HA-treated sperm, relative to control and CS- or DS-treated sperm. HP and HA treatments increased the levels of capacitated and acrosome-reacted sperm over time, compared to other treatment groups (p<0.05). In addition, sperm exposed to HP or HA for 1 h before IVF exhibited significantly improved fertilizing ability, as assessed by 2 pronucleus (PN) formation and cleavage rates at d 2. Exposure to these GAGs also enhanced in vitro embryo development rates and embryo quality, and increased the ICM and total blastocyst cell numbers at d 8 after IVF (p<0.05). A real-time PCR analysis showed that the expression levels of pluripotency (Oct 4), cell growth (Glut 5), and anti-apoptosis (Bax inhibitor) genes were significantly higher in embryos derived from HA- or HP-treated sperm than in control or other treatment groups, while pro-apoptotic gene expression (caspase-3) was significantly lower in all GAG treatment groups (p<0.05). These results demonstrated that exposure of bovine sperm to HP or HA positively correlates with in vitro fertilizing ability, in vitro embryo developmental potential, and embryonic gene expression.

• 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.

• Asian-Australasian Journal of Animal Sciences
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• 제19권5호
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• pp.632-637
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• 2006

Oviduct-specific glycoprotein 1 (OVGP1) is implicated in playing a role in fertilization and early embryo development. In this study, we have obtained the sequence of intron 9 of OVGP1 gene in swine. Comparative sequencing of Meishan (a native Chinese breed) and Large White pig breeds revealed an A/T substitution at position 943. A PCR-EcoRI-RFLP assay was developed to detect this mutation. Polymorphism analysis in Qingping animals showed that pigs with BB genotype had lower number of piglets born alive (NBA) in multiple parities than pigs with AA (p<0.05) and AB genotype (p<0.01). In Large $White{\times}Meishan$ ($LW{\times}M$) $F_2$ offspring, the weight of both ovaries (OW) of the BB genotype was significantly lighter than that of AB (p = 0.05) and AA (p<0.01) genotypes. Analysis of the data also revealed that the mutation locus affected these two traits mostly by additive effects. These studies indicated that the polymorphism was associated with NBA and OW in two distinct populations and further investigations in more purebreds or crossbreds are needed to confirm these results.

• 한국수정란이식학회지
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• 제28권3호
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• pp.163-168
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• 2013

Embryo transfer (ET) technology is of high importance in modern cattle breeding programs. ET is one step in the process of removing one or more embryos from the reproductive tract of an outstanding donor female and transferring them to one or more recipient females. Embryos also can be produced in the laboratory via techniques such as in vitro fertilization (IVF). But the actual transfer of an embryo is only one step in a series of processes that may include some or all of the following: superovulation and insemination of donors, collection of embryos, isolation, evaluation and short-term storage of embryos, micromanipulation and genetic testing of embryos, freezing of embryos and embryo transfer. Cryopreservation and direct transfer of frozen-thawed embryos is common-place with pregnancy rates near that of fresh embryos. Polymerase chain reaction (PCR) technology is currently being used for sexing embryos, and this technology will be used for "embryo diagnostics" and "embryo genomics" in the future. Although, many limitations and problems remain to overcome, these and other new technologies promise to change livestock breeding drastically in the next decade.

• 한국임상수의학회지
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• 제31권6호
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• pp.539-543
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• 2014

질 종양은 개 암컷생식기에서 두 번째로 많이 발생하는 종양 중 하나이며, 대부분은 평활근종으로 보고되고 있다. 질 종양의 기원은 질의 내벽 또는 외벽 유래로 구분되며, 성호르몬의 영향을 받아 성장하는 경향이 있다. 본 증례는 질 외벽 유래의 종양으로 종양은 모두 복강 내 존재했으며, 방광 및 직장, 특히 요관과 같은 주변장기와의 유착이 확인되었다. 종양은 탐색적 개복술을 통해 주변장기의 손상 없이 제거했으며 중성화 수술 또한 동시에 시행되었다. 제거된 종양은 조직학적으로 평가 한 결과 질 평활근종으로 확인되었다.

• 한국동물생명공학회지
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• 제37권4호
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• pp.285-291
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• 2022

In 1951, Colin Russell Austin and Min Chueh Chang identified "capacitation", a special process involving ejaculated spermatozoa in the female reproductive tract. Capacitation is a phenomenon that occurs in vivo, but almost all knowledge of capacitation has been obtained from in vitro studies. Therefore, numerous trials have been performed to establish in vitro capacitation methods for various studies on reproduction. Although a series of studies have been conducted to develop an optimal protocol for inducing capacitation, most have focused on identifying the appropriate chemical compounds to induce the capacitation of boar spermatozoa in vitro. Therefore, the purpose of this study was to identify the optimal incubation time for inducing capacitation in vitro. Duroc semen was incubated for various periods (60, 90, and 120 min) to induce capacitation. Sperm function (sperm motility, motion kinematic parameters, and capacitation status) was evaluated. The results showed that total sperm motility, rapid sperm motility, progressive sperm motility, curvilinear velocity, and average path velocity significantly decreased in a time-dependent manner. However, the capacitation status did not show any significant changes. Taken together, these results indicate that an incubation time of more than 60 min suppresses sperm motility and motion kinematic parameters. Therefore, we suggest that 60 min may be the best incubation time to induce capacitation without negative effects on sperm motility and motion kinematics in boar spermatozoa in vitro.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.52-52
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• 2003

Granulocyte-macrophage colony stimulating factor (GM-CSF) is synthesized in the female reproductive tract and has been shown to play an important role in human and murine embryo development and implantation. However, the mechanism of GM-CSF on the embryo development is unknown. Recent studies suggested that GM-CSF may be increase the expression of implantation relented genes, such as interleukin-1 (IL-1) system. Our aim of this study was to compare the interleukin-1$\alpha$ (IL-1$\alpha$), interleukin-1$\beta$ (IL-1$\beta$) and interleukin-1 receptor antagonist (IL-lra) mRNA between the GM-CSF supplemented group and control group in mouse embryos. Mouse 2-cell embryos were cultured in P-1 medium supplemented with or without mouse GM-CSF (10 ng/ml). The number of total and apoptotic cell in blastocyst were assessed by TUNEL. And then, the expression of IL-1$\alpha$, IL-1$\beta$ and IL-1ra mRNA in blastocyst were examined by RT-PCR.

• Anatomy and Cell Biology
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• 제57권1호
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• pp.119-128
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• 2024

Glucocorticoids play a physiologic role in the adult male reproductive functions, modulating gonadal steroid synthesis and spermatogenesis, through the glucocorticoid receptor (GR). The expression of GR has been described in several key testicular cell types, including somatic cells and early germ cell populations. Nothing is known on GR in human spermatozoa. Herein, we explored the GR expression and its possible role in normal and testicular varicocele semen samples from volunteer donors. After semen parameter evaluation by macro- and microscopic analysis, samples were centrifuged; then spermatozoa and culture media were recovered for further investigations. By western blotting and immunofluorescence analyses we evidenced for the first time in spermatozoa the presence of GR-D3 isoform which was reduced in sperm from varicocele patients. By treating sperm with the synthetic glucocorticoid dexamethasone (DEXA), we found that survival, motility, capacitation, and acrosome reaction were increased in both healthy and varicocele samples. GR involvement in mediating DEXA effects, was confirmed by using the GR inhibitor mifepristone (M2F). Worthy, we also discovered that sperm secretes different cortisol amounts depending on its physio-pathological status, suggesting a defence mechanism to escape the immune system attach in the female genital tract thus maintaining the immune-privilege as in the testis. Collectively, our data suggests a role for glucocorticoids in determining semen quality and function, as well as in participating on sperm immune defensive mechanisms. The novelty of this study may be beneficial and needs to take into account in artificial insemination/drug discovery aimed to enhancing sperm quality.

• Applied Microscopy
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• 제29권2호
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• pp.241-253
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• 1999

북부 온대지방에 서식하는 박쥐는 그 대부분이 동면을하며, 그 생식유형은 특이하여 가을에 교미하여 사정된 정자는 자성생식도내에 긴 동면기간 동안 저장되어 있는 것으로 알려져 있다(예: 한국큰관박쥐). 환언하면 저장되어 있던 정자는 이듬해 봄에 배란되는 난자와 수정하여 초여름에 출산하게 된다. 본 연구는 월동하는 정자 두부 Plasma membrane의 성분과 활성 등 형태적 및 기능적 특성을 관찰 분석함으로써, 인공임신율을 높일 수 있는 보다 효과적인 한냉온도 정자저장방법을 모색하기 위하여 시도되었다. 동면하는 한국큰관박쥐와 햄스타 및 비동면동물(마우스)의 정자 Plasma membrane이, 실온과 한냉온도(박쥐-동면온도)에서, 생존율, 첨체반응을, 단백질분포, 능동수송에 관여되는 효소$(Na^+-K^+-ATPase)$의 활성 및 주사전자현미경적 조직화학상을 분석하였다. 본 연구에서 얻은 실험결과에 의거하면, 동면동물인 한국큰관박쥐와 햄스타 및 비동면동물인 마우스 정자의 생존율과 첨체반응율은, 실온과 한냉(박쥐-동면)온도간에 의의있는 차이를 발견할 수 없었고, 단백질 분포와 $Na^+-K^+-ATPase$ 효소활성 및 주사전자현미경적 조직화학적 소견에 있어서도 두 온도간에 의의있는 차이는 없었다. 이와 같이 실온과 한냉온도 간에 형태적 기능적 차이가 없다고 하는 사실은, 동면동물이든 비동면동물이든 Spermatozoa plasma membrane이 동면중 또는 동면 이후에 안정적인 수정을 보장해 주는 형태적 및 기능적 보호구조임을 시사해 준다.

• 한국수정란이식학회지
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• 제22권4호
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• pp.245-249
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• 2007

The aim of present experiment was to examine hatching rate as in vitro indicator of viability of porcine embryos before early stage embryo transfer such as zygotes or 2-cell stage embryos. Cumulus-oocyte complexes (COCs) collected from ovaries were matured in North Carolina State University 23 (NCSU-23) containing 10% porcine follicular fluid (pFF), 10 ng/ml epidermal growth factor (EGF), $10{\mu}g/ml$ follicle stimulating hormone (FSH), $35{\mu}g/ml$ luteinizing hormone (LH), and 1mg/ml cysteine. After 24 hours, the COCs were transferred to the same medium without hormones. After 65h of maturation, oocytes were exposed to phosphate buffered saline (PBS) with 7% ethanol (v/v) for 7 minutes, and then the oocytes were washed and cultured in tissue culture medium (TCM) 199 containing 5 ug/ml cytochalasin B for 5h at $38.5^{\circ}C$ in an atmosphere of 5% $CO_2$ and 95% air with high humidity. After cytochalasin B treatment, the presumptive parthenotes were cultured in porcine zygote medium (PZM)-5 and cleavage of the parthenotes was assessed at 72h of activation, Normally cleaved parthenotes were cultured for an additional 8 days to evaluate their ability to develop to blastocyst and hatching stages. The fetal bovine serum (FBS) were added at Day 4 or 5 with concentrations of 2.5, 5 or 10%. The blastocyst rates were ranged within $39.1{\sim}70%$ in each treatment. However hatching rate was dramatically decreased in non-addition group. In this experiment, embryo viability in female reproductive tract may be estimated before embryo transfer with in vitro culture adding FBS by hatching ability.