• The Korean Journal of Malacology
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• v.26 no.1
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• pp.51-62
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• 2010

The ultrastructures of germ cells and the accessory cells during spermatogenesis and mature sperm ultrastructure in male Gomphina veneriformis, which was collected on the coastal waters of Yangyang, East Sea of Korea, were investigated by transmission electron microscope observations. The morphology of the spermatozoon has a primitive type and is similar to those of other bivalves in that it contains a short midpiece with four mitochondria surrounding the centrioles. Accessory cells are observed to be connected to adjacent germ cells, they contain a large quantity of glycogen particles and lipid droplets in the cytoplasm. Therefore, it is assumed that they are involved in the supplying of the nutrients for germ cell development, while any phenomena associated with phagocytosis of undischarged, residual sperms by lysosomes in the cytoplasm of the accessory cells after spawning was not observed in this study. The morphologies of the sperm nucleus type and the acrosome shape of this species have a cylindrical and modified long cone shape, respectively. In particular, the axial filaments in the lumen of the acrosome, and subacrosomal granular materials are observed in the subacrosomal space between the anterior nuclear fossa and the beginning part of axial filaments in the acrosome. The spermatozoon is approximately $50-55{\mu}m$ in length including a long sperm nucleus (about $7.80{\mu}m$ in length), an acrosome (about $1.13{\mu}m$ in length) and tail flagellum ($40-45{\mu}m$). The axoneme of the sperm tail flagellum consists of nine pairs of microtubules at the periphery and a pair at the center. The axoneme of the sperm tail shows a 9+2 structure. Some charateristics of sperm morphology of this species in the family Veneridae are (1) acrosomal morphology, (2) the number of mitochondria in the midpiece of the sperm,. The axial filament appears in the acrosome as one of characteristics seen in several species of the family Veneridae in the subclass heterodonta, unlikely the subclass pteriomorphia containing axial rod instead of the axial filament. As some characteristics of the acrosome structures, 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 belong to the family Veneridae in the subclass heterodonta, unlikely 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. The number of mitochondria in the midpiece of the sperm of this species are four, as one of common characteristics appeared in most species in the family Veneridae.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.21 no.9
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• pp.57-63
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• 2020

To develop simple acrosome staining of horse spermatozoa, this study tested the binding properties of Coomassie brilliant blue G or R on the sperm smears after 3.7% paraformaldehyde (PF) or 35% methanol (MT) fixation. After being fixed with PF and stained with 0.05, 0.1, or 0.2 % of CBB G or R for 2 min, horse spermatozoa were examined for their intact acrosome status. The intact acrosome of fresh horse spermatozoa were 62.6% and 61.5% with 0.05% of the G and R CBB solution, but 80.2 and 79.7% with G type and 78.1 and 76.0% with R type. On the other hand, when MT was used for fixation, the acrosome reacting sperm ratio was 3.5%, but was 9.0% in the case of PF. These results show that the intact acrosome of horse sperm could be judged using a 0.1~0.2% CBB G or R staining technique. PF would be an essential fixative for examining acrosome reacting horse spermatozoa. This method could be used to identify sperm with a damaged acrosome during low-temperature storage or cryopreservation for artificial insemination of horses.

• Applied Microscopy
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• v.28 no.4
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• pp.603-613
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• 1998

The present study was designed in order to observe relationship between the endoplasmic reticulum and the Golgi complex during spermiogenesis of the long-fingered bat (Miniopterus schreibersi fuliginosus). The testes were obtained from adult bats and treated with the prolonged osmification or fixed with ferrocyanide reduced osmiun. In the Golgi phase, The Golgi complex shows an oval shape, and was composed of a cortex and a medullar enclosing acrosome. The Golgi vacuoles with electron-dense granules of crescent shape were fused with each other. The smooth endoplasrnic reticulum was scattered in all the area of the cytoplasm. In the cap phase, The Golgi complex was crescent in shape, and faced to a nucleus. Large and small vesicles were fused with each other, and then fused with a acrosomal vacuole. The rough endoplasmic reticulum was close to the large Golgi vacuole. In the acrosome phase, The Golgi complex was moved to behind of the acrosome face. Small vesicles were fused with an acrosome, and cisternae of the trans-face of Golgi complex was connected with an acrosome in the early acrosome phase. The smooth endoplasmic reticulum was distributed in the cytoplasm. The annulate lamellar was originated from a radial body-annulate lammellae complex. In the maturation phase, The Golgi complex with dilated cistrern appeared in the cytoplasm, and also, annulate lamellar was observed in the cytoplasm. The connection of the annulate lamellar with the cistern of radial body suggests that an annulate lamellar seems to be closely related to radial body. The smooth endoplasmic reticulum was scattered in the cytoplasm in the early Golgi phase, but annulate lamellar-radial body complex which might be a residual and disappearing form of the smooth endoplasmic reticulum appeared in the acrosome phase. The Golgi complex steadily remained in the late maturation phase when the endoplasmic reticulum began to disappear from the cytoplasm: the Golgi complex was still occurred after acrosome formation. The observations obtained in the present study, which was characterized by the presence of the Golgi complex in the late maturation phase, suggests that the Golgi complex may play an important role also even after the acrosome formation.

• Korean Journal of Animal Reproduction
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• v.15 no.1
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• pp.1-6
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• 1991

This study was carried out to investigate the effects of Ca, BSA, heparin, semen storage and individual bull on motility and acrosome reaction of bovine fresh sperm and sperm stored in lactose-egg yolk solution(LES) at 5$^{\circ}C$ for 4hours, and the results obtained were as follows: 1. When sperm was incubated in SCS containing Ca, BSA, Ca + BSA, heparin, heparin + Ca, heparin + BSA, and heparin + Ca + BSA for 15 minutes, there was significant difference in sperm motility among the treatments, especially BSA showed significantly higher sperm motility than the others. Also there was significant difference in sperm acrosome reaction among the treatments, especially BSA and Ca + BSA showed significantly higher sperm acrosome reaction than the others. 2. Bull KNC 1 showed significantly higher sperm motility than KNC 1, HOL 1 and 2 in both fresh and stored semen, however KNC 1 showed significantly lower sperm acrosome reaction than KNC 1, HOL 1 and 2. Therefore, there was significant difference in sperm motility and acrosome reaction among individual bulls. 3. When KNC 1 and KNC 2 sperm were incubated in SCS and SCS + Ca, SCS + BSA, SCS + Ca + BSA, SCS + heparin, SCS + heparin + Ca, SCS + heparin + BSA, and SCS + heparin + Ca + BSA, there was significant difference in sperm motility among individual bulls, especially BSA in KNC 1 and BSA, Ca and Ca + BSA in KNC 2 showedsignificantly higher motility than the others. However, there was significant difference in sperm acrosome reaction among individual bulls, Ca in KNC 1 and Ca + BSA in KNC 2 showed higher acrosome reaction than the others.

• Clinical and Experimental Reproductive Medicine
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• v.24 no.3
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• pp.311-318
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• 1997

Bypassing acrosome reaction and fusion process in intracytoplasmic sperm injection(ICSI), most of injected spermatozoa still contain intact acrosome contents and plasma membrane. It Is not known yet what acrosome contents and plasma membrane of spermatozoa have effect on the development of embryo. For further understanding of fertilization process after ICSI, we studied the time of pronucleus formation, disappearance and first cleavage in human zygote, and pregnancy rate in relation to acrosome reaction rate of spermatozoa after ICSI. Seventy cycles undergoing ICSI program were randomly selected. Sperm suspension from 38 cycles were treated 50% human follicular fluid(hFF) for 3 hours in order to induce acrosome reaction, others were not treated as control. Acrosome reaction in hFF treated and non-treated group was assessed by fluorescein isothiocyanate(FITC)-conjugated Arachis hypogea(PNA) and Pisum sativum agglutinin(PSA). Oocytes were classified into 'good' and 'poor' according to their morphology. After ICSI, fertilization of oocytes were assessed by detection of two pronuclei at 16 hours. The pronuclei disappearance and first cleavage of zygotes were observed at 24 hours, and then embryos were transferred to uterus after culture for 72 hours. The rate of acrosome reaction of spermatozoa in hFF treated group was significantly higher than that in control(p<0.01). Fertilization rates of good oocytes were not different both control and hFF treated group(81.3%(174/206) vs. 72.1%(102/130)). But, in poor oocytes, the fertilization rates in hFF treated group(72.1%(149/183)) were increased compared than those of control group (63.6%(98/140), p<0.01). In either good or poor oocytes, the rates of pronuclei disappearance in hFF treated-spermatozoa injected oocytes were higher than control (59.1%(103/174), 56.4%(84/149) vs. 32.4%(33/102), 37.8%(37/98), p<0.01). Also, the rates of thirst cleavage were increased in hFF treated group (31%(54/174), 24.1%(36/149)) compared than those of control group (10.8%(11/102), 13.2%(13/98), p<0.01). The pregnancy rates of hFF treated group (42.1%(16/38)) were slightly higher than control group (28.1%(9/32), p>0.05). But, the pregnancy rate of group which possessed more than one cleavaged zygote at 24 hours was higher than group which did not (45.2%(19/42) vs. 21.4%(6/28), p<0.05). From these results, the development of zygotes were faster in higher acrosome reacted sperm group than lower acrosome reacted sperm group after ICSI. Our results may be explained that acrosomal membrane and plasma membrane are easily detached from spermatozoa in acrosome reacted spermatozoa compared with acrosome intact sperm in the cytoplasm of oocyte during pronuclear formation. We conclude that the injection of acrosome reacted spermatozoa will increase the pregnancy rate as they can induce fast embryonic development in ICSI.

• Korean Journal of Animal Reproduction
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• v.3 no.2
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• pp.24-31
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• 1979

The maturation changes in morphology were studied with the spermatozoa collected from the testis and three successive parts of the epididymis in Korean native Cattle. Acrosomal granules were observed in the testis. Avoiding the cap and acrosome phases, the tail base and the striated column of the neck were formed in spermatides. The volume of the acrosome was decreased during transit from the testis to the epididymis. The cell membranes were also separated from the acrosome or damage during the spermatozoan passage through successive parts of the reproductive tract. Cytoplasmic droplets were observed in the spermatozoa collected from various parts of the reproductive tract.

• Development and Reproduction
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• v.1 no.2
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• pp.133-139
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• 1997

To study the function and structure of Golgi apparatus in the spermiogenesis of long-fingered bat (Miniopterus schreibersi fuliginosus), the testis obtained from adult bat was treated with the prolonged osmification or fixed with ferrocyanide reduced osmium. golgi apparatus was oval shape in early Golgi phase, and was composed of cortex and medullar enclosing acrosome in mid Golgi phase. The vesicles of crescent shape Golgi apparatus were closed or fused with small or large vesicles at the periphery of acrosome. Golgi apparatus moved behing the acrosome face in cap phase, but the Golgi apparatus was still active. According to this, Golgi apparatus appears to be involved in the formation of acrosome and sperm tail. Transfer of materials from Golgi to acrosme seems to be carried out not only by fusion of large vesicles with acrosomal vesicles but also by detachment of coated vesicle from various cisternae of Golgi fusing with acrosomal vesicle.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.6
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• pp.748-756
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• 2000

Effects of xanthine (X), xanthine oxidase (XO), and catalase (C), $H_2O_2$, and carbohydrates on sperm capacitation, acrosome reaction, and fertilizing ability in vitro were examined. Glucose alone, but not fructose, supported the maximum rate of sperm capacitation and acrosome reaction. However, in the combination of X, XO, and C (XXOC) or $H_2O_2$, fructose alone also supported maximum capacitation, acrosome reaction, and fertilization. Either insufficient or excessive amounts of $H_2O_2$ decreased sperm capacitation and the acrosome reaction. In order to understand how glucose generates $H_2O_2$ or other reactive oxygen species in sperm cells, 6-aminonicotinamide, an inhibitor of the pentose-phosphate pathway (PPP), and apocynin, an inhibitor of NADPH oxidase, were added to sperm suspensions in glucose-containing medium. Results appeared that sperm capacitation, acrosome reaction, and fertilization were consequently inhibited by either one of these compounds. These inhibitory effects were nullified by addition of XXOC. These results support the hypothesis that glucose, in addition to being a substrate for glycolysis, facilitates sperm capacitation and the acrosome reaction by generating reactive oxygen species through G-6-P dehydrogenase and NADPH oxidase.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.11
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• pp.1553-1558
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• 2002

This study was carried out to obtain informations regarding the effect of N-acetyl-D-glucosamine in the LEY (lactoseegg yolk) diluent according to incubation time in 5 ml maxi-straw and the effects of freezing rate, thawing temperature and thawing time in the LEN (lactose-egg yolk and N-acetyl-D-glucosamine) diluent on acrosome morphology and motility of frozen-thawed boar sperm. The study showed that the LEN diluent was higher post-thaw NAR (normal apical ridge) acrosome than the LEY diluent for 0.5 h incubation at 37$^{\circ}C$. However, there were no differences between the LEN and LEY diluents on post-thaw sperm motility according to incubation time. The straws frozen from 5.0 cm (20$^{\circ}C$/min) to 17.0 cm (1$^{\circ}C$/min) above the liquid nitrogen surface did not show any significant differences on post-thaw sperm motility. However, the straws frozen above 5.0 cm from the liquid nitrogen surface were higher NAR acrosome than those frozen above 17.0 cm. The post-thaw percentages of motile sperm and NAR acrosome were significantly higher (p<0.05) for the maxi-straws submerged for 40 or 45 sec in a 52$^{\circ}C$ water bath than for 30, 35, 50 or 55 sec. The mean sample temperatures of maxi-straws after 40 or 45 sec submersion were 20.7 or 26.4$^{\circ}C$. In conclusion, the sample temperature of the thawed semen was very important for post-thaw sperm survival in the LEN diluent of 5 ml maxi-straw. When the temperature of the thawed semen was 20.7$^{\circ}C$, the percentages of motile sperm and NAR acrosome were highest.

• Korean Journal of Animal Reproduction
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• v.15 no.3
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• pp.189-193
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• 1991

This study was carried out ot investigate effects of X-537A on hydrogen ion concentration insperm washed solution and sperm acrosome reaction. The results obtained were as follows. 1. When bovine sperm was twice washed with SHP solutions of pH 6.8 and 7.4 and again washed with SHP solution containing 4$\mu$M of X-537A, in case of pH 6.8 the sperm washed with 4$\mu$M of X-537A showed signifciantly(p<0.01) higher hydrogen ion concentration in sperm washed solution than the sperm washed without X-537A. 2. When the sperm was twice washed with SHP solution and then washed with SHP solution containing 4$\mu$M of X-537A, sperm acrosome rection rate was signifciantly(p<0.01) increased from 12min after incubation in the sperm washed without X-537A, but was signifciantly(p<0.01) increased from 8 min after incubation in the sperm washed with 4$\mu$M of X-537A. 3. When the sperm was twice washed with SHP solution and then washed with SHP solution containing 0, 4 and 40$\mu$M of X-537A, and then incubated in m-TALP for 120 min, sperm acrosome reaction rate was significantly(p<0.01) increased from 15 min after incubation in 0, 4 and 40$\mu$m OF X-537A. However at 60 min incubation 40$\mu$M of X-537A showed significantly(p<0.01) higher sperm acrosome reaction rate than 0 and 4$\mu$M and at 120 min incubation 4 and 40$\mu$M of X-537A showed signifciantly(p<0.01) higher acrosome reaction rate than 0$\mu$M of X-537A.