• Journal of Embryo Transfer
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• v.29 no.1
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• pp.59-66
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• 2014

Transcription activator like effector nucleases (TALENs) are artificial restriction enzymes generated by fusing a TALE DNA binding domain to a DNA cleavage domain which remove and introduce specific genes to produce transgenic animals. To investigate the efficient laboratory techniques for the injection of TALEN mRNA, pEGFP-N1 commercial plasmid were microinjected into porcine parthenogenetic and in vitro fertilization (IVF). In Experiment 1, to investigate injection time, compared 4 different time durations (2 hr, 4 hrs, 6 hrs & 8 hrs) after post activation of parthenogenetic embryos and after 6 hrs of co-incubation with sperms in IVF embryos. There were significant difference (P<0.05) in development to the blastocysts (4.4, 8.9, 3.9, 0.6%), GFP expression in blastocysts (1.3, 5.7, 2.3, 0.0%) which injected after post activation of 4 hrs compared with other 3 groups. IVF embryos after 2 hrs and 4 hrs injected were expressed GFP significantly higher than rest of two groups (P<0.05). In Experiment 2, compared development of 2 different concentrations ($20ng/{\mu}l$ and $50ng/{\mu}l$) of EGFP injection. There were significant difference (P<0.05) between two treatments which has higher cleavage (58.8 vs 41.9%), blastocysts development rate (13.0 vs 11.1%) and GFP expressed blastocysts (5.7 vs 0.0%) in $20ng/{\mu}l$ than the $50ng/{\mu}l$ in parthenogenetic embryos. In IVF embryos, only $20ng/{\mu}l$ injected embryos were expressed GFP (4.2%) after 7 days of incubation and 77.3 vs 64.7% of cleavage, 26.4 vs 23.5% development to blastocysts. In Experiment 3, three different volumes (5, 10 and 20 pl) were microinjected into porcine embryos to determine the most appropriate volume. Out of 3 groups, significantly higher development rates of cleavage (68.3, 58.0, 29.3%), blastocysts (11.7, 12.7, 0.5%) and GFP expressed blastocysts (2.9, 7.8, 0.0%) were shown in the 10 pl group (P<0.05). In conclusion, these results imply that $20ng/{\mu}l$ concentration, 10 pl of volume and injection at 4 hrs after post activation for parthenogenetic and 2~4 hrs after IVF, $20ng/{\mu}l$ concentration and 10 pl volume for IVF embryos were more effective microinjection conditions.

• Journal of Embryo Transfer
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• v.19 no.2
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• pp.75-80
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• 2004

This study was performed to examine the cytotoxic effects of nicotine on the seminiferous tubules and Leydig cells in mice testis. A different amount of nicotine (2～15 mg/kg, for two weeks, one dose of 100 mg/kg) were administered to four-month male mice, and then the optical microscopic findings of its effect on testis of the mice are as follows: 1. The group that 2 mg/kg of nicotine was administered showed normal findings that nucleus and cytoplasm of Leydig cells are distinct, while the other group that 5 mg/kg of nicotine was given to showed nucleus and cytoplasm are swollen and thickened a little, and slightly dyed. 2. The group that 10 mg/kg of nicotine was given had irregular arrangement of spermatogenesis inside seminiferous tubules so it was impossible to distinguish phages of seminiferous tubules. It was also impossible to observe cells due to fusion of their nucleuses, and distinct cytoplasm. 3. The group that 15 mg/kg of nicotine was administered showed destruction of nucleuses and cytoplasm of spermatocytes and sperms and a fill of fibered connective tissues so that it is impossible to observe rumens of seminiferous tubules.

• Journal of Aquaculture
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• v.8 no.4
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• pp.295-306
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• 1995

The present study was performed to produce supermales and superfemales in Oreochromis niloticus by combination of induced sex reversal and diploid gynogenesis. More than $95\%$ of female was obtained by oral administration of $17\beta-estradiol$ (480 mg/kg diet) to the eutheroembryonic larvae of this species. The result of progeny tests with XY pseudofemales showed that incidences of male progeny were ranged from $71.4\%\;to\;73.7\%$. p.opo.lion of male from XY pseudofemale was not significantly different (P>0.05) from that of $\chi^2$ (1:3), but it was significantly different (P<0.01) from that of $\chi^2$(1:1) Gynogenetic diploids were produced by applying cold shocks 3 min after insemination to the eggs of XY pseudofemale sperms were genetically inactivated by ultraviolet rays of 4,050 $erg/mm^2$. Fertilization rates of gynogenetic diploids were not different from that of their controls, however, hatching rates and early survival rates were silghtly lower than those of controls. In the cytogenetic studies, there was no difference in the size of cell and nucleus between the gynogenetic diploids and the controls. At 80 days after hatching, frequencies of gynogenetic male were higher than those of gynogenetic female (P<0.01) in their population.

• Korean Journal of Environmental Biology
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• v.27 no.4
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• pp.323-333
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• 2009

Parabens are alkyl esters of p-hydroxybenzoic acid, which are widely used in foods, cosmetics, and pharmaceutic products as preservatives. Absorbed parabens are metabolized fastly and excreted. Actually human body is exposed to complex mixture of parabens. Safety assessment at various toxicological end points revealed parabens have a little acute, subacute and chronic toxicities. Some reports have argued that as parabens have estrogenic activity, they are associated with the incidence of breast cancer through dermal absorption by cosmetics. There is an inference that antiandrogenic activity of parabens may give rise to a lesion of male reproductive system, but also there is an contrary. At cellular level, parabens may inhibit mitochondrial function of sperms and androgen production in testis, but also there is an contrary. Parabens seem to have little or no toxicity in embryonic development. Parabens can cause hemolysis, membrane permeability change in mitochondria and apoptosis, suggesting cellular toxicity of parabens. Parabens evoked endocrine disruption in several fish species and have toxic effect on small invertebrates and microbes. Therefore, the toxicity of parabens should be considered as a potentially toxic chemical in the freshwater environment. In conclusion, though parabens may be considered as a low toxic chemical, more definite data are required concerning the endocrine disrupting effect of parabens on human body and aquatic animals according to route and term of exposure as well as the residual concentration of parabens.

• Reproductive and Developmental Biology
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• v.38 no.3
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• pp.115-121
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• 2014

Humulus japonicus is an ornamental plant in the Cannabaceae family. Although the mode of action of Humulus japonicus is not fully understood, a strong relationship was observed between anti-inflammatory and anticancer in some types of cells. Recent studies also have shown that Humulus japonicus possesses anti-inflammatory activities and may significantly improve antioxidant potential in Raw 264.7 macrophage cells. Thus, the aim of this study was evaluated the effect of Humulus japonicus extract on sperm motility and subsequent preimplantation developmental competence of the bovine embryos. After in vitro maturation, the oocytes with sperms were exposed in in vitro fertilization (IVF) medium supplemented with Humulus japonicus extract (0.01, 0.05, $0.1{\mu}g/mL$, respectively) for 1 day. In our results, exposure of IVF medium to Humulus japonicus extract did not affect sperm motility and percentage of penetrated oocytes but ROS intensity was significantly decreased by $0.01{\mu}g/mL$ compared with other groups (p< 0.05). Moreover, treatment with $0.01{\mu}g/mL$ of Humulus japonicus extract was higher the frequency of blastocyst formation than the any other groups (p<0.05). Otherwise, treatment with $0.01{\mu}g/mL$ of Humulus japonicus extract not increased the total cell number but reduced apoptotic-positive nuclei number. In conclusion, our results indicate that supplementation of Humulus japonicus extract in IVF medium may have important implications for improving early embryonic development in bovine embryos.

• 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.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.31 no.2
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• pp.210-216
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• 1998

Relationship between population group properties and docosahexaenoic acid (DHA) content of the Pacific oyster had been carried out at the Bukman oyster farm near Tongyeong based on the regular monthly sampling from November 1994 through April 1996. DHA content in lipids was least abundant in June and most abundant in November. Minimum DHA content close to 'zero' in lune suggests that much of DHA are being used for maturing and discharging of eggs and sperms, in consideration of the fact that lure is the spawning periods of the oyster. The corelationships among DHA content, mortality, and growth coefficient, have not been recognized. The approximate positive relations have been acknowledged between DHA content and the individual density in times of harvest, and also the individual weight. But it seems that the relaltions between DHA content and the individual weight are not directly related, rather it seems that it is the result from the population density effects caused by the relations between DHA and the number of individuls. But the meaning of the above mentioned relations can not be clearly defined yet.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.25 no.5
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• pp.413-431
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• 1992

Gonadal development, fertilization and egg development in the maternal body and reproductive cycle of ovoviviparous rockfish, Sebastes inermis, were investigated histologically. Gonadosomatic index(GSI) of male and female were increased from September and reached maximum values in December. In the male, GSI decreased from January, but in the female maintained high values till February and decreased from March. Hepatosomatic index(HSI) was related to GSI conversely. In both sex, HSI increased from February and reached maximum in August as the gonad were degenerating and resting, and began to decrease from September as gonad were glowing. This ovoviviparous rockfish copulates in December. Fertilization with sperms maintained between ovulated oocytes in the ovary occurs in January mainly. Egg development in the ovarian cavity and discharging of hatched preiarva occurs from January to February. The reproductive cycle includes the successive stages: Growing(September), Mature (October-November), Ripe and Fertilization(Decembr-Janua), Egg development and Discharging of hatched larva(January-February), Degeneration and Resting(February-August). According to the frequency distribution of egg diameter and histological observation, the ovoviviparous rockfish discharged the prelarva at a time in a spawning season. The sexual maturation is first attained at 2 ages. All females and males reaches first maturity at body length of 17.1cm and 15.1cm respectively. The mean number of the embryos increased with the increase of the total length of female.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.28 no.5
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• pp.539-548
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• 1995

The process of yolk absorption and the growth pattern of churn salmon, Oncorhynchus keta alevin were studied. Matured males and females of chum salmon were sampled from the Namdaechun River in the east coast of Korea, and then an artificial fertilization was carried out using their eggs and sperms. Fertilized eggs were incubated in controlled water temperature of $13^{\circ}C$ and samples of 40 alevins were randomly taken at just hatching and at every three days between 12 days alter hatching and yolk absorption. The means of total length and total weight of hatched alevins were respectively 1.97cm and 1.85g and the yolk absorption took about 33 days after hatching, based on the morphological measurement. Growth curves of total length, total weight and somatic weight were fitted well with the Gompertz growth model. However, the shrinking in yolk-sac length, yolk-sac weight, yolk-sac height and yolk-sac volume revealed a linear phenomenon until the absorption of yolk was completed. The relative growth of chum salmon alevin, such as the relationship between total length and total weight, or between total weight and yolk-sac weight, was also studied.

• The Korean Journal of Malacology
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• v.29 no.2
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• pp.105-111
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• 2013

The gonadal development of triploid and diploid Pacific abalones, Haliotis discus hannai was histologically investigated in spawning season. Diploid abalones had matured oocytes and spermatozoa, but most triploid had spermatocytes or developing oocytes that was slightly retarded in gonadal development compared to diploid abalones. In spawning experiment of triploid and diploid abalones, spawning rates of diploid male and female were 100%, but those of triploid female was 50% and male was 25% respectively. Investigation of spawned abalone eggs and spermatozoa revealed that length of diploid sperms head were 17.47 ${\mu}m$, breadth of head were 10.31 ${\mu}m$, length of spermatozoa were 130.72 ${\mu}m$, but those of triploid spermatozoa were 11.83 ${\mu}m$, 7.89 ${\mu}m$ and 103.36 ${\mu}m$ respectively. Triploid spermatozoa were significantly small to diploid spermatozoa (p < 0.05). The eggs of diploid and triploid were not different in size. The cross experiment between oocytes produced by triploids and spermatozoa by diploids ($3n{\times}2n$ cross) revealed that no fertilization were occurred, and $2n{\times}3n$ cross also revealed same result.