• 한국발생생물학회지:발생과생식
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• 제3권1호
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• pp.87-93
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• 1999

생쥐 정자의 수정능력획득과 첨체반응에 작용하는 $Na^{+}$/H$^{+}$ antiporter의 역할을 조사하고자 하였다. $Na^{+}$/H$^{+}$ antiporter를 특이적으로 억제하는 dimethyl amiloride는 정자의 자발적인 첨체반응을 농도 의존적으로 억제한 반면 난포액 및 calcium ionophore인 A23l87에 의해 유도된 첨체반응은 억제하지 못하였다. 이러한 결과는 정자내 $Na^{+}$/H$^{+}$ antiporter에 의한 1가이온의 출입과 이에 따른 세포질내 pH 조절이 정자의 수정능력 획득과 자발적인 첨체반응에 조절요인으로 작용함을 암시한다. 수정능력을 획득한 정자에서 난포액 등의 agonist 또는 A23l87에 의해 유도되는 첨체반응은 $Na^{+}$/H$^{+}$ antiporter와는 무관하게 진행되는 것으로 사료된다.

• Clinical and Experimental Reproductive Medicine
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• 제27권1호
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• pp.9-14
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• 2000

Objective: The sperm acrosome reaction is a $Ca^{2+}$-dependent exocytotic event that is triggered by adhesion to the mammalian egg's zona pellucida. Previous studies suggested a role of $Ca^{2+}$ channels in acrosome reactions. This study was conducted to investigate the T-type calcium channel is operated in acrosome reaction of human spermatozoa. Method: Human semen samples were obtained from healthy donors with normal criteria. The spermatozoa were divided into five groups: Group 1 were non-treated as a control; Group 2 where spermatozoa were exposed to 5 ${\mu}M$ $Ca^{2+}$ A23187 $(Ca^{2+}i)$; Group 3 where spermatozoa were exposed 5 ${\mu}M$ $Ca^{2+}i$ and mibefradil; Group 4 where spermatozoa were exposed 5 ${\mu}M$ $Ca^{2+}i$ and nifedipine, and Group 5 where spermatozoa were treated with 5 ${\mu}M$ $Ca^{2+}i$ and both of mibefradil and nifedipine. Spermatozoa in all groups were retrieved after incubation for 15 and 30 minutes at $37^{\circ}C$. After staining with PSA-FITC, fluorescence was observed under a fluorescence microscope, and AR was evaluated on a total>100 spermatozoa/side. Result and Conclusion: We observed on acrosome reaction inhibition rate in human spermatozoa the various of concentration of mibefradil, nifedipine. Maximum response was noted with 1.0 ${\mu}M$ mibefradil and the decrease of acrosome reaction inhibition rate 45%. Nifedipine in acrosome reaction inhibition rate was only about 25%. The $Ca^{2+}i$-induced AR of spermatozoa was significantly suppressed by mibefradil. Incidence of the suppression was depending on concentration of mibefradil. Results from the present study suggest that the human spermatozoa possess T-type channel. The observation that reversible inhibitor of T channels in male germ cells provides a new mechanism of contraceptive action.

• Asian-Australasian Journal of Animal Sciences
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• 제19권4호
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• pp.486-494
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• 2006

In order to protect the spermatozoa against cold shock, hen egg yolk is widely used as a cryoprotective agent in semen freezing extenders for domestic animals. The protective action of yolk is largely presumed to be due to low density lipoproteins (LDL). The effects of LDL on sperm quality of bull and northern pike (Esox lucius) after freezing-thawing have been reported, but no study has been made to evaluate the effect of LDL on boar sperm motility and other characteristics. The experiment was carried out to investigate the effect of LDL on the freezing of boar sperm in 0.25 ml straws. The aim was to evaluate the quality of boar spermatozoa cryopreserved in the presence of LDL. Motility of semen cryopreserved in LDL was analyzed and compared to semen cryopreserved with Tris-citric acid-glucose (TCG) and Tris-citric acid-fructose (TCF), two basic freezing extenders containing egg yolk. Similarly, acrosome and plasma membrane integrity were also evaluated and compared to semen cryopreserved with TCG and TCF. Analysis of sperm quality after freeze-thaw showed that the motility, acrosome and plasma membrane integrity were improved with LDL in the extender, as compared to the TCG and TCF. The highest post-thaw integrity of acrosome and plasma membrane and motility were obtained with 9% LDL (w/v). Consequently, the optimum LDL concentration in the extender was 9%. It is also suggested that the concentration of LDL addition is important for the effect on boar sperm protection during freezing and thawing. The percentage of motile spermatozoa was significantly higher after freezing in 9% LDL than in TCG and TCF 54.4% versus 30.4% and 30.1% (p<0.05), respectively. The integrity of acrosome and plasma membrane were also significantly higher at 70.3% and 50.5% respectively with semen frozen in 9% LDL extender compared to TCG at 37.8% and 30.3% and TCF at 36.4% and 29.9%, respectively (p<0.05),. In conclusion, we propose that extender containing LDL extracted from hen egg yolk could be used as a cryoprotective media with a better efficiency than TCG and TCF. LDL improved boar semen quality, allowing better spermatozoa motility, acrosome and plasma membrane integrity after the freeze-thaw process. Furthermore, we found out that the extender with 9% LDL concentration significantly enhanced motility, acrosome and plasma membrane integrity of boar sperm after freezing and thawing.

• Asian-Australasian Journal of Animal Sciences
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• 제29권9호
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• pp.1247-1255
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• 2016

Heparin binding proteins (HBPs) are produced by accessory glands. These are secreted into the seminal fluid, bind to the spermatozoa at the time of ejaculation, favour capacitation, acrosome reaction, and alter the immune system response toward the sperm. The present study was conducted with an objective to assess the effect of purified seminal plasma-HBPs (SP-HBPs) on cross bred cattle bull sperm attributes during two phases of cryopreservation: Pre freezing and freezing-thawing. SP-HBPs were purified from pooled seminal plasma by heparin affinity chromatography. Three doses of SP-HBPs i.e. 10, 20, $40{\mu}g/mLs$ semen were standardized to find out the optimum dose and $20{\mu}g/mLs$ was found to be an optimum dose. Semen as such and treated with SP-HBPs was diluted with sodium citrate-egg yolk diluter and cryopreserved as per the standard protocol. Sperm parameters i.e. motility, viability, Hypo-osmotic swelling test (HOST), acrosome damage, in vitro capacitation and lipid peroxidation were evaluated in SP-HBP treated and untreated (control) semen at both phases of cryopreservation. A considerable variation in percent sperm motility, viability, membrane integrity (HOST), acrosome damage, acrosome reaction and lipid peroxidation was observed at both phases among the bulls irrespective of the treatment. Incubation of neat semen with $20{\mu}g/mL$ SP-HBP before processing for cryopreservation enhanced the average motility, viability, membrane integrity by 7.2%, 1.5%, 7.9%, and 5.6%, 6.6%, 7.4% in pre-frozen and frozen-thawed semen in comparison to control. There was also an average increase of 4.1%/3.9% in in vitro capacitation and acrosome reaction in SP-HBPs-treated frozen-thawed semen as compared to control. However, binding of SP-HBPs to the sperm declined acrosome damage and lipid peroxidation by 1.3%/4.1% and 22.1/$32.7{\mu}M$/$10^9$ spermatozoa in SP-HBP treated pre-frozen/frozen-thawed semen as compared to control, respectively. Significant (p<0.05) effects were observed only in motility, HOST and in vitro acrosome reaction. It can be concluded that treatment of neat semen with SP-HBPs before cryopreservation minimized the cryoinjury by decreasing the generation of reactive oxygen species.

• 한국발생생물학회지:발생과생식
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• 제17권2호
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• pp.121-132
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• 2013

Ultrastructural characteristics of the germ cells and accessory cells in testis during spermatogenesis and taxonomic values of mature sperm morphology of Ruditapes philippinarum were investigated by the transmission electron microscope and scanning electron microscope observations. The testis is the diffuse organ that consists of branching acini containing developing germ cells and accessory cells associated with spermatogenesis. The morphology of the spermatozoon is of the primitive type and is somewhat different to those of other bivalves. The morphologies of the sperm nucleus type and the acrosome shape of this species have a cylinderical type and a modified cone shape, respectively. As some ultrastructural characteristics of the acrosomal vesicle, the peripheral parts of two basal rings show electron opaque part, while the apex part of the acrosome shows electron lucent part. These characteristics of sperm belong to the family Veneridae in the subclass Heterodonta, unlike a characteristic of the subclass Pteriomorphia showing all part of the acrosome being composed of electron opaque part. In particular, a cylinder-like nucleus of the sperm is curved. The spermatozoon is approximately $48-51{\mu}m$ in length, including a long acrosome (about $2.4{\mu}m$ in length), a curved sperm nucleus (about $3.40{\mu}m$ in length), and a tail flagellum. The axoneme of the sperm tail shows a 9+2 structure.

• 한국수정란이식학회지
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• 제26권4호
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• pp.287-296
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• 2011

Mammalian fertilization is a complex cascade process consisting of sperm migration through the female reproductive tract, physiological changes to sperm such as sperm capacitation and acrosome reaction, and sperm-egg interaction in the oviduct in vivo. On the other hand, in vitro fertilization (IVF) is a process by which egg cells are fertilized by sperm outside the body: in vitro. IVF has been used for a variety of purposes in reproductive biotechnology for human and animals. The discovery of sperm capacitation in 1951 promoted the development of IVF technology. In the initial stage of IVF, sperm capacitation in preincubation medium was shown to be essential to fuse with eggs. Besides, sperms should detour some of the in vivo regulations for IVF. This review introduces a general mammalian fertilization process, including sperm capacitation, removal of cumulus matrix, acrosome reaction, and sperm-egg fusion and focuses on the roles of key biochemical molecules, signal mechanisms, and genes involved during IVF and novel results of sperm-oocyte interaction elucidated in various gene-knockout mice models.

• 한국발생생물학회지:발생과생식
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• 제15권3호
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• pp.239-247
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• 2011

Spermatogenesis and taxonomic values of mature sperm morphology of in male Septifer (Mytilisepta) virgatus were investigated by transmission electron microscope observations. The morphologies of the sperm nucleus and the acrosome of this species are the cylinder shape and cone shape, respectively. Spermatozoa are approximately 45-50 ${\mu}m$ in length including a sperm nucleus (about 1.26 ${\mu}m$ long), an acrosome (about 0.99 ${\mu}m$ long), and tail flagellum (about 45-47 ${\mu}m$). Several electron-dense proacrosomal vesicles become later the definitive acrosomal vesicle by the fusion of several Golgi-derived vesicles. The acrosome of this species has two regions of differing electron density: there is a thin, outer electron-dense opaque region (part) at the anterior end, behind which is a thicker, more electron-lucent region (part). In genus Septifer in Mytilidae, an axial rod does not find and also a mid-central line hole does not appear in the sperm nucleus. However, in genus Mytilus in Mytilidae, in subclass Pteriomorphia, an axial rod and a mid-central line hole appeared in the sperm nucleus. These morphological differences of the acrosome and sperm nucleus between the genuses Septifer and Mytilus can be used for phylogenetic and taxonomic analyses as a taxonomic key or a significant tool. The number of mitochondria in the midpiece of the sperm of this species are five, as seen in subclass Pteriomorphia.

• Fisheries and Aquatic Sciences
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• 제10권3호
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• pp.143-149
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• 2007

This study describes spermatogenesis and sperm ultrastructure of the granular ark, Tegillarca granosa using light and electron microscopy. In the active spermatogenic season, the testis comprises many spermatogenic follicles that contain germ cells in different developmental stages. Primary spermatocytes in the pachytene stage are characterized by synaptonemal complexes. The early spermatids are characterized by the appearance of several Golgi bodies, increased karyoplasmic electron density, and tubular mitochondria. The mass of proacrosomal granules consists of numerous heterogeneous granules with high electron density that are about 20 nm in diameter. From the midstage of spermiogenesis, the well-developed mitochondria in the cytoplasm aggregate posterior to the nucleus and surround the proximal and distal centrioles. The proacrosomal granules condense and form a single acrosome with a thin envelope. During late spermiogenesis, the acrosome begins to elongate becoming conical. The sperm is approximately $35.0{\mu}m$ long and consists of a head, midpiece, and tail. The head comprises a round nucleus and a conical acrosome. A micro fibrous axial rod is observed between the nucleus and acrosome. The midpiece has a calyx-like structure with five mitochondria, and the tail, which has the typical "9+2" microtubular system, originates from the distal centriole.

• 한국가축번식학회지
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• 제14권4호
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• pp.309-313
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• 1990

This stduy was carried out to investigate the effect of concentration of PC12 and washing of sperm of sperm motility and acrosome reaction, and the effect of sperm incubated in mTALP solution containing PC12 112.5$\mu\textrm{g}$/ml on development of follicular oocytes matured in vitro. The results obtained were as follows. 1. When fresh sperm was once washed and then incubated in mTALP solution containing 0, 75, 112.5 and 225$\mu\textrm{g}$/ml PC12 for 15minutes, 225$\mu\textrm{g}$/ml showed significantly higher percent of acrosome reacted sperm than 0, 75, 112.5 and 225$\mu\textrm{g}$/ml. 2. When once or twice washed fresh sperm was cultured in mTALP containing 112.5$\mu\textrm{g}$/ml PC12 for 15minutes, no-washing showed significantly higher percent of motile sperm than that once- and twice-washing showed significantly higher percent of acrosome reacted seprm than no- and once-washing. 3. When sperm was cultrued in mTALP containing PC12, 112.5$\mu\textrm{g}$/ml for 15minutes and then was cocultured with bovine follicular oocytes matured in vitro, 11.2 to 22.4% of the oocytes were coleaved to more than 2cell stage.

• Advances in nano research
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• 제11권6호
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• pp.649-655
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• 2021

Nanotechnology is widely considered a major technology of the twenty-first century. Nanoparticles (NPs) has been shown to pass through reproductively significant biological barriers such as the blood-testicle and placental barriers. Thus, the purpose of this study was to determine the effect of silver Nanoparticles (Ag-NPs) on sperm-egg interaction and spermatozoa quality parameters in quail spermatozoa. Semen was suspended in Ringer solution containing Ag-NPs levels at 5.5 × 106 sperm/ml (0, 0.01, 0.1, 1 and 10 ppm). The results indicated that when sperm were counted at 0.1 ppm, the number of holes formed on the inner perivitelline layer was significantly increased compared to the control. The 10 ppm group had a significant reduction in sperm viability. At 0.1 and 1 ppm, the membrane integrity was significantly decreased (P < 0.05). All treatments (except 0.01 ppm Ag-NPs) had a significant (P < 0.05) effect on the percentage of spermatozoa with an intact acrosome when compared to the control group. At 0.1, 1, and 10 ppm Ag-NPs, morphological defects in the acrosome were observed. As a result, Ag-NPs is likely capable of destroying the acrosome membrane. This research indicates that Ag-NPs may be cytotoxic to spermatozoa by impairing sperm functionality and increasing sperm mortality.