• Korean Journal of Ichthyology
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• v.19 no.4
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• pp.286-291
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• 2007

The bitterling, Acheilognathus koreensis spermatozoon has been examined by electron microscopy. The epididymal spermatozoa of A. koreensis are representing typical characteristic of cyprinid spermatozoa including the lateral insertion of flagellum, the organization of centriolar complex in shallow nuclear fossa and the asymmetrical arrangement of mitochondria. The sperm mid-piece contains a large mitochondrion characteristic enclosed by membranous vesicles. The mitochondria aspect is different from that of other cyprinid spermatozoa, which their mitochondria have a conventional aspect and never fuse to form a mitochondrial derivative. In term of sperm evolution, the fused mitochondria are regarded as the apomorphic character in comparison with the separate mitochondria. The single mitochondrion is not found in cyprinid spermatozoon except for Rodeus and Pungtungia.

• 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.

• Clinical and Experimental Reproductive Medicine
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• v.29 no.1
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• pp.57-66
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• 2002

Objective : This study was designed to investigate the interrelationship and clinical usefulness of sperm morphology by strict criteria (SM), acrosome reaction following ionophore challenge test (ARIC) and sperm penetration assay (SPA) using zona-free hamster ova as prognostic factors in in vitro fertilization. Materials and Methods: Semen samples were provided by 83 patients undergoing IVF. We first evaluated the differences between normal fertilization group and poor fertilization group on three andrologic tests. Secondly, we analyzed the relationship between the three andrologic tests and in vitro fertilization on IVF settings. Finally, we evaluated the effectiveness of the three andrologic tests as the prognostic indicators for fertilizing ability. Results: The fertilization rate of all men in the poor fertilization group was less than 30%; but there was no evidence that this poor fertilization was due to oocyte defects. The results of three andrologic tests were significatly higher in normal fertilization group. Fertilization rate (%) in vitro was highly correlated (p<0.001) with % normal sperm by SM, ARIC value (%), and SPA result. By using Receiver-Operator-Characteristic curve (ROC), we evaluated the effectiveness of these three tests. The sensitivity and specificity of SM, ARIC test and SPA in predicting fertilization potential in IVF setting were 76% and 75%, 84% and 90%, and 76% and 95%, respectively. Conclusion: Our data suggest that the three andrologic tests can be reliable tools as prognostic factors of sperm fertilizing ability. Among these test, ARIC test and SPA gave more accurate information on fertilizing capacity. ARIC test was shown to have a predictive value for fertilizing ability comparable to that of SPA that appears to be a simple and cost-effective addition to current andrology laboratory. Combined application of these three tests may give more information on predicting sperm fertilizing capacity.

• Journal of Embryo Transfer
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• v.33 no.1
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• pp.17-22
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• 2018

This study was conducted to determine the effect of pentoxifylline levels on sperm motility, survival rate, sperm membrane integrity of frozen semen and fresh-extended equine semen in Jeju cross-bred horses. As a result of sperm characteristic comparison depending on pentoxifylline levles at 30 minutes post-thaw, the progressive motilities were $53.25{\pm}2.87$ (4mM pentoxifylline) and $50.28{\pm}2.14$ (8mM pentoxifylline) and significantly higher compared to the control group($40.09{\pm}5.15$) and other treatment group (16mM pentoxifylline, $41.27{\pm}2.82$). The progressive fast motility were $22.44{\pm}1.62$ (4mM pentoxifylline,) and $22.74{\pm}3.07$ (8mM pentoxifylline) and significantly higher compared to the control group ($13.47{\pm}1.48$) and other treatment group (16mM pentoxifylline, $14.66{\pm}3.68$) (p<0.05). As a result of sperm characteristic comparison depending on pentoxifylline levles at 30 minutes post-thaw were $68.96{\pm}1.64$ (4mM pentoxifylline) and $67.90{\pm}6.72$ (8mM pentoxifylline) and significantly higher compared to the control group ($53.48{\pm}4.84$) and other treatment group (16mM pentoxifylline, $58.14{\pm}2.65$) (p<0.05). In conclusion, these results suggest that treatment groups with 4mM and 8mM pentoxifylline were higher compare to equine seperm mobility and the control group and treatment groups with more than 16mM pentoxifylline has a negative effect on sperm characteristics. After thawing, the total motility in post-thawed equine sperm has increased by 10 percent for 1 hour. these results suggest that pentoxifylline contributes to the improvement of the equine sperm motility and characteristics in post-thawed semen.

• Journal of Embryo Transfer
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• v.29 no.2
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• pp.163-169
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• 2014

The objective of this study was to evaluate the characteristics of Korean Native Cattle sperm frozen-thawed with L-cysteine and/or catalase. The semen from bulls was collected by the artificial vagina method, and Triladyl containing 20% egg-yolk and/or L-cysteine (L), catalase (C) and L-cysteine + catalase was added to the diluted semen for cryopreservation. The results showed that sperm viability was significantly higher in the L-cysteine + catalase ($69.49{\pm}3.16%$) group than in the control ($60.5{\pm}3.94%$) group (p<0.05). Acrosome damage was significantly lower in the L-cysteine ($17.12{\pm}1.08%$) group than in the control ($21.46{\pm}1.14%$), catalase ($20.54{\pm}0.76%$), and L-cysteine + catalase ($19.29{\pm}0.65%$) groups (p<0.05). In addition, the level of intact mitochondria in the spermatozoa was significantly higher in the L-cysteine ($58.65{\pm}1.39%$) group than in the control ($50.63{\pm}2.37%$) group (p<0.05). The hydrogen peroxide level in the frozen-thawed sperm was significantly lower in the L-cysteine ($3.74{\pm}1.66%$), catalase ($4.65{\pm}1.87%$), and L-cysteine + catalase ($8.11{\pm}2.15%$) groups than in the control ($13.22{\pm}1.6%$) group (p<0.05). The glutathione level was significantly higher in the L-cysteine ($1.33{\pm}0.03%$) group than in the control ($1.08{\pm}0.06%$), catalase ($1.05{\pm}0.02%$) and L-cysteine + catalase ($1.11{\pm}0.03%$) groups (p<0.05). In conclusion, L-cysteine and catalase could protect the membrane of Korean Native Cattle sperm from damage during sperm cryopreservation. Especially, L-cysteine was more effective for keeping acrosomes and mitochondria intactness during sperm cryopreservation.

• Clinical and Experimental Reproductive Medicine
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• v.47 no.1
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• pp.61-67
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• 2020

Objective: In this study, specimens from testicular biopsies of men with nonobstructive azoospermia (NOA) were used to investigate whether RNF8 gene could serve as a biomarker to predict the presence of sperm in these patients. Methods: Testicular biopsy specimens from 47 patients were classified according to the presence of sperm (positive vs. negative groups) and investigated for the expression of RNF8. The level of RNF8 gene expression in the testes was compared between these groups using reverse-transcription polymerase chain reaction. Results: The expression level of RNF8 was significantly higher in testicular samples from the positive group than in those from the negative group. Moreover, the area under the curve of RNF8 expression for the entire study population was 0.84, showing the discriminatory power of RNF8 expression in differentiating between the positive and negative groups of men with NOA. A receiver operating characteristic curve analysis showed that RNF8 expression had a sensitivity of 81% and a specificity of 84%, with a cutoff level of 1.76. Conclusion: This study points out a significant association between the expression of RNF8 and the presence of sperm in NOA patients, which suggests that quantified RNF8 expression in testicular biopsy samples may be a valuable biomarker for predicting the presence of spermatozoa in biopsy samples.

• Journal of Embryo Transfer
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• v.30 no.1
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• pp.7-15
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• 2015

The objective of this study was to investigate the efficiency of nicotinic acid on sperm cryosurvival and fertilization ability in frozen-thawed boar semen. Boar semen was collected by glove-hand method and was frozen using freezing solution treated to 0, 5, 10 and 20 mM of nicotinic acid. The frozen sperm for sperm characteristic analysis was thawed such as viability, acrosome reaction, and mitochondrial integrity. The frozen-thawed sperm was estimated by SYBR14/PI double staining for viability, FITC-PNA/PI double staining for acrosome reaction and Rhodamine123/PI double staining for mitochondrial integrity using a flow cytometry. The embryo was estimated in vitro development and DCFDA staining for reactive oxygen species assessment. As results, frozen-thawed sperm viability was significantly higher in 5 and 10 mM ($61.1{\pm}1.5%$,$64.7{\pm}2.0%$) of nicotinic acid than other groups (0 mM, $52.1{\pm}2.3%$; 20 mM, $47.8{\pm}5.1%$, P<0.05). The live sperm with acrosome reaction was significantly higher in 5 and 10 mM of nicotinic acid ($26.1{\pm}1.8%$, $24.9{\pm}1.5%$) than other groups (0 mM, $35.3{\pm}0.8%$; 20 mM, $36.5{\pm}1.9%$, P<0.05). The live sperm with mitochondrial integrity was significantly higher in 5 and 10 mM ($84.2{\pm}3.6%$, $88.4{\pm}2.3%$) of nicotinic acid than other groups (0 mM, $77.3{\pm}4.4%$; 20 mM, $73.3{\pm}3.6%$, P<0.05). Blastocyst rate of in vitro development was significantly higher in 10 mM ($17.0{\pm}1.3%$) of nicotinic acid than other groups (0 mM, $9.4{\pm}0.5%$; 5mM, $12.6{\pm}0.8%$; 20 mM, $5.0{\pm}1.0%$, P<0.05). Moreover, total cell number was higher in 5 and 10 mM ($53.6{\pm}2.9%$, $57.9{\pm}2.8%$) of nicotinic acid than other groups (0 mM, $41.0{\pm}1.4%$; 20 mM, $23.2{\pm}2.8%$, P<0.05). Hydrogen peroxide in embryos was lower in 5 mM nicotinic acid ($0.7{\pm}0.1%$) than other groups (0 mM, $1.0{\pm}0.1%$; 10mM, $0.9{\pm}0.0%$; 20 mM, $1.4{\pm}1.0%$, P<0.05). In conclusion, nicotinic acid-treated semen improves cryosurvival and quality of spermatozoa. Also, the fertilized oocytes with nicotinic acid improve quality of embryo and blastocyst formation.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.12
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• pp.1721-1727
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• 2008

Four experiments were conducted to study the following: i) the influence of different concentrations of sucrose (0.15, 0.3 and 0.5 M with osmolality of 308, 500 and 760 mOsm/kg, respectively) in diluents and control diluent (370 mOsm/kg) on intensity of motility and progressive motility of goat sperm without rehydration and freezing step in four incubation periods (0, 0.5, 2 and 4 h after dilution); ii) the influence of gradual dilution (in 3 steps) on improvements in ascertained results of the first experiment; iii) cryoprotective effects of different concentrations of sucrose (0.15, 0.22, 0.29 and 0.37 M with osmolality of 450, 560, 740 and 920 mOsm/kg, respectively) plus 7% glycerol and 20% egg yolk in basic diluent (Tris-Citric acid-Fructose) and iv) the effect of two concentrations of sucrose (0.15 and 0.22 M) with and without glycerol (7%). In experiment 1, we obtained better results for control diluent, 0.15 and 0.3 M sucrose supplemented diluents with 0 and 0.5 h incubation periods. In experiment 2, apart from a slight improvement, similar tendencies to experiment 1 were observed. In experiment 3, we obtained the best result for diluent with 0.22 M sucrose with regard to intensity of motility, progressive motility, live sperm and normal acrosomes ($40{\pm}4%$, $3.1{\pm}0.2$, $37{\pm}4%$ and $37{\pm}4%$, repectively). In experiment 4, we obtained the best result for diluent with 0.22 M sucrose plus 7% glycerol in regard to intensity of motility, progressive motility and live sperm ($39{\pm}3%$, $3.6{\pm}0.4$ and $41{\pm}4%$, respectively). The characteristic normal acrosomes in diluents without glycerol, i.e. diluents with 0.15 and 0.22 M sucrose showed better results ($39{\pm}8$ and $42{\pm}6%$ respectively). With regard to the release of hyaluronidase enzyme there were no significant differences between diluents (p>0.05). The results of the diluents with 0.15 and 0.22 M sucrose without glycerol were slightly lower than those with glycerol ($69{\pm}11$ and $70{\pm}11$ vs. $72{\pm}11$ and $70{\pm}11{\times}120{\times}10^6$ units $ml^{-1}$, respectively). In conclusion, the use of concentrated sucrose solutions showed that goat sperm can tolerate osmolality up to 560 mOsm (0.22 M) in the freezing period. In addition, glycerol proved to be a necessary cryoprotective agent in the cryopreservation of goat sperm, particularly for intensity of motility, progressive motility and live sperm.

• Korean Journal of Animal Reproduction
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• v.26 no.1
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• pp.53-59
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• 2002

This study were performed to investigate the effect of magnetic field exposure on semen characteristic and the weights of body, reproductive organs and liver, kidney and spleen in mice. In magnetic field exposure for 15 days, sperm concentrations and viability were significantly lower in magnetic field(15.7$\times$10$^{6}$ $m\ell$, 29.3%) than that in control group(25.1$\times$10$^{6}$ $m\ell$, 34.4%)(P＜0.05). The proportion of sperm abnormality were significantly increased in magnetic field exposure groups for 15 days than that in control group(P＜0.05). The exposure of magnetic field in mice didn't affect the body and reproductive organ weight such as testis, epididymis, vasicular gland and coagulatin gland. The weight in liver and kidney were not affect in magnetic field exposure groups. However, the spleen weight were significantly higher(P＜0.05) in group exposed with than without magnetic field. This studies indicate the short or long term magnetic field exposure in mice were noxious effects on the sperm characteristics and spleen weight, but didn't affect body, reproductive organs, and liver and kidney weight.

• Reproductive and Developmental Biology
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• v.33 no.1
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• pp.25-28
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• 2009

This study was carried out to investigate the general characteristics, such as volume, pH, sperm motility and sperm concentration of the semen collected from Beagle dogs (age $24{\sim}48$ months, weight $10{\sim}15\;kg$) by using the method of digital manipulation of the penis, and the effect of preservation temperature and time on motility of fresh semen. Multiple ejaculates were collected from 4 male Beagles. The average volume, pH, motility and sperm concentration of the second fraction (contained with small volume of the third fraction) per ejaculation were $2.94{\pm}0.24(SD)\;ml$, $6.43{\pm}0.42(SD)$, $97.04{\pm}3.50(SD)%$ and $1.67{\pm}0.23(SD){\times}10^8\;cells/ml$, respectively. Average semen volume per ejaculate, semen pH, sperm motility and sperm concentration of the first fraction from the ejaculate were $1.24{\pm}0.20(SD)\;ml$, $6.03{\pm}0.26(SD)$, $1l.30{\pm}4.02(SD)%$ and $7.25{\pm}1.02(SD){\times}10^5\;cells/ml$. Those of second fraction were $2.52{\pm}0.32(SD)\;ml$, $6.32{\pm}0.31(SD)$, $96.25{\pm}3.52(SD)%$ and $2.35{\pm}0.35(SD){\times}10^8\;cells/ml$. Those of third fraction were $2.71{\pm}0.27\;(SD)\;ml$, $6.52{\pm}0.20(SD)$, $95.65{\pm}2.78(SD)%$ and $5.72{\pm}0.29(SD){\times}10^7\;cells/ml$. Motility of semen was higher at $17^{\circ}C$ preservation temperature than $5^{\circ}C$ or $36^{\circ}C$ during preservation period. When preservation temperature was $17^{\circ}C$, motility was $96.54{\pm}2.05(SD)%$ at 1 h, $90.20{\pm}3.90(SD)%$ at 6 h, $89.05{\pm}2.01(SD)%$ at 12 h, $78.21{\pm}3.50(SD)%$ at 18 h, $45.24{\pm}6.25\;(SD)%$ at 24 h and $30.75{\pm}17.24(SD)%$ at 30 h, respectively.