• Journal of the Korea Academia-Industrial cooperation Society
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• v.22 no.4
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• pp.504-512
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• 2021

Sperm formation disorders and sperm quality degradation comprises the largest proportion of male infertility caused by TCDD. To solve this problem, this study examined the effects of Gobonyangjeonbang oriental medicine prescription on the endocrine function and reproductive toxicity-related indicators in rat-induced TCDD-induced reproductive. Male SD rats were divided into five groups of seven animals and tested. The normal control group was administered the vehicle and saline, the TCDD alone group was administered intraperitoneally with TCDD (2 ㎍/kg, weeks) and physiological saline, and the test group was administered orally by dividing GYB (75, 150, and 300 mg/kg) into three concentrations for six weeks. Weight loss was observed in all groups administered TCDD. Regarding the hormonal changes, a significant decrease in free testosterone was observed in the GYB 300 mg/kg group (p<0.01). In addition, some of the germ cell destruction, seminiferous tubular atrophy, and decrease in sperm count was improved in a concentration-dependent manner in the testicular tissue of the GYB-treated group. In addition, Johnsen's score and serotoli cell index (SCI) were improved in a concentration-dependent manner (p<0.05). Overall, GYB can be used in drug therapy rather than medical procedures to solve male infertility in the future.

• Journal of Aquaculture
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• v.7 no.4
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• pp.189-205
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• 1994

Formerly, adult-tiger puffer, Takifugu rubripes with ova caught in the sea, were used for seedling production. But it was difficult to secure naturally-ripened adults. For the purpose of adult tiger puffer in captivity, this study was carried out. To determine the growth 220 tiger puffers hatched in 1990 (3-year-old) and 1991 (2-year-old) were used. For spawning and egg incubation leading to fry development, eggs were stripped from tiger puffers hatched in 1988 (5-year-old) and 1990 (3-year-old) through human chorionic gonadotropin (BCG) treatments. In May, 1993, mean body length and mean body weight of 2-year-old tiger puffer were $30.72\pm1.35cm\;and\;1,048\pm228 g,$ and that of 3-year-old tiger puffers were $36.02\pm1.17cm$ and $1,402\pm66g$ respectively. The relationship between body length (L) and body weight (W) of 2-year-old the tiger puffers during the experiment period was represented as $W\;=\;1.7892L^{31524}\times10^5$ (r= 0.9436) and that of 3-year-old, $W=\;3.2840L^{36099}\times10^6$ (r= 0.9070) respectively. The GSI in female 2-year-old-fish changed from $0.23\times0.l2\;to\;0.74\pm0.08$, during the experiment period, and in male it didn't change remarkably until November, but thereafter it increased and showed a peak of $8.69\pm5.09$. The GSI of 3-year-old-fish showed a peak of $8.05\pm5.58$ in April in female and $12.65\pm4.60$ in May in male. The change of HSI in 3-year-old-fish was correlative to the change of GSI, but in 2-year-old-fish it was little correlative. In female gonad of 2-year-old tiger puffer, the mature oocytes reached $350{\mu}m$ in April, but thereafter they didn't spawn and became atrophied. But in male gonad, a great number of spermatozoa were crowded in the testicular lobuli in April. Female gonad of 3-year-old tiger puffer had the mature oocytes of 650 pm in March and the ripe oocytes, $900{\mu}m$ in April. Male testis development was similar to that of 2-year-old-fish. Egg-stripping after hormone treatments was possible past 139 hours and 142 hours from each of two 5-year-old-fish (500IU/kg, BW), and after 114 hour from a 3-year-old-fish (1,000 IU/kg, BW) under water temperature $16.3\~17.8^{\circ}C$. Eggs stripped amounted was 650 g and 400 g from two 5-year-old-fish and 610 g from the 3-year-old-fish, and fertilization rates were $98.0\%,\;97.4\%\;and\;96.5\%$ respectively. All the hatched larvae devloped into normal fry.

• Development and Reproduction
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• v.13 no.2
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• pp.105-114
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• 2009

Ethane 1,2-dimethane sulfonate (EDS) is a well-known alkylating agent used as selective Leydig cell (LC) toxicant to create a testicular dysfunction model. Previous studies including our own clearly demonstrated the dramatic weight loss of the androgen dependent accessory sex organs such as epididymis, seminal vesicle and prostate gland in this 'LC knock-out' rats. The present study was performed to evaluate the effect of EDS administration on histological changes of the epididymis, seminal vesicle and prostate in adult rats. Adult male Sprague-Dawley rats (350$\sim$400 g B.W.) were injected with a single dose of EDS (75 mg/kg, i.p.) and sacrificed on weeks 0, 1, 2, 3, 4, 5, 6 and 7. Tissue weights (testis, epididymis, seminal vesicle and prostate gland) were measured. The histological changes of tissue were observed by a light microscopy using hematoxylin & eosin staining. Weights of the reproductive and accessory organs progressively declined after the EDS treatments (weeks 1, 2 and 3). After this, the decrease was stopped, then gradually returned to the normal levels. There was a partial (about 60%) recovery of the epididymis weight during weeks $6{\sim}7$. The cross section of epididymis revealed an increase in thickness of the epithelium during weeks $1{\sim}3$. In contrast, considerable reduction of epithelial thickness in seminal vesicle was observed during same period. Similarly, a reduction in thickness of prostate epithelial layer was found during weeks $1{\sim}3$, then it was back to normal thickness after week 4. Taken together, the present study demonstrated that the temporally induced androgen-deficiency by EDS treatment could result the prominent alterations in histology of the accessory sex organs. Further studies on the physiological and molecular regulation of these androgen-sensitive organs using EDS model will be helpful to understand the normal and pathological development and differentiation mechanism of these organs.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.22 no.5
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• pp.266-280
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• 1989

Gonadal maturation of the Korean pomfrets, Pampus echinogaster (Basilewsky) and Pampus argenteus (Euphrasen) were histologically investigated based on the samples captured in the East China Sea from January 1987 to December 1988. Gonadosomatic index (GSI) of P. echinogaster began to increase from March, and reached maximum between May and July. It began to decrease from July and reached mini-mum between August and February. P. argenteus had a similar cycle, however, P. argenteus has higher values in April than P. echinogaster. Hepatosomatic index (HSI) were positively related to GSI. HIS of P. echinogaster and P. argenteus reached maximum in $April\~July$ and $April\~August$, respectively, Fatness coefficient of two Pampus species were low in the summer, and high in the winter. Ovary is of saccular structure, and testis is of lobular structure. From February, the early oocyte (ca. $100\mu$ in diameter grows) rapidly at the germinal epithelium of ovarian sacs. From March to April the oocytes grew up to cu $400\~500\mu$ in diameter. At this stage, the yolk globules are accumulated rapidly in the cytoplasmic layer. From May, the oocytes roached ca. $650\~850\mu$ in diameter, and they are spawned in $May\~July$. After spawning the residual follicles and remained ripe eggs degenerate. From February, spermatogonia grows into spermatocyte on the epithelium of the testicular lobuli. From May, spermatozoa appeared and spawning occurs. After spawning, the epithelium is thickened and the remained spermatozoa degenerate. Annual reproductive cycle of two Pampus species could be divided into four successive stages: Growing stage ($March\~April$), Mature stage ($April\~May$), Ripe and spent stage ($June\~July$) and Recovery and resting stage ($August\~January$). Absolute fecundity of P. echinogaster was $9,441\~135,294$, and that of P. argenteus was $50,678\~221,894$. Absolute fecundity of two Pampus species were positively related to body length and total weight. Relative fecundity was positively related to body length, while it was reversely related to total weight. The increasing rate of absolute fecundity of P. echinogaster was lower than P. argenteus. In P. echinogaster half of female and male reached first maturity at body length of $15.0\~$17.9cm and $12.0\~14.9cm$, respectively. All of females and males reached first maturity at body length of $18.0\~20.9cm$ and $21.0\~23.9cm, respectively. In P. argenteus all of females and males reached first maturity at body length of 18.6cm and 16.7cm$, respectively.

• Toxicological Research
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• v.33 no.2
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• pp.107-118
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• 2017

Although alternative test methods based on the 3Rs (Replacement, Reduction, Refinement) are being developed to replace animal testing in reproductive and developmental toxicology, they are still in an early stage. Consequently, we aimed to develop alternative test methods in male animals using mouse spermatogonial stem cells (mSSCs). Here, we modified the OECD TG 489 and optimized the in vitro comet assay in our previous study. This study aimed to verify the validity of in vitro tests involving mSSCs by comparing their results with those of in vivo tests using C57BL/6 mice by gavage. We selected hydroxyurea (HU), which is known to chemically induce male reproductive toxicity. The 50% inhibitory concentration ($IC_{50}$) value of HU was 0.9 mM, as determined by the MTT assay. In the in vitro comet assay, % tail DNA and Olive tail moment (OTM) after HU administration increased significantly, compared to the control. Annexin V, PI staining and TUNEL assays showed that HU caused apoptosis in mSSCs. In order to compare in vitro tests with in vivo tests, the same substances were administered to male C57BL/6 mice. Reproductive toxicity was observed at 25, 50, 100, and 200 mg/kg/day as measured by clinical measures of reduction in sperm motility and testicular weight. The comet assay, DCFH-DA assay, H&E staining, and TUNEL assay were also performed. The results of the test with C57BL/6 mice were similar to those with mSSCs for HU treatment. Finally, linear regression analysis showed a strong positive correlation between results of in vitro tests and those of in vivo. In conclusion, the present study is the first to demonstrate the effect of HU-induced DNA damage, ROS formation, and apoptosis in mSSCs. Further, the results of the current study suggest that mSSCs could be a useful model to predict male reproductive toxicity.

• Journal of Aquaculture
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• v.11 no.4
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• pp.529-546
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• 1998

Gonadal part that developed by indifferentiation period for 6 months after hatching is made as gonad and fat body. These gonad are thin semi-transparant and undistinguished germ cell. Germinal epithelium is distinguished by development of gonad epithelial tissue from 7 months after hatching. Sex differentiation is begun by oogonia develoment at 8 months after hatching. Primary oocytes grow over germinal epithelium of gonadal cavity, at 9 months after hatching, gonadal cavity become ovarian cavity as they increasing. As soon as oocytes at 13 months after hatching are filled with the whole part of gonad, degeneration of oocyte is begun. And then, gonad has cavity tissue, a small number of oocyte are located in gonadal cavity. At 15 months after hatching, new primary oocyte develop and cavity of ovarian tissue in the central of ovarian cavity. Spermatogonia multiplicate and cavity tissue consist of testicular tissue. These gonad become hermaphrodite and then ditermine the sex of female and male. These results show the red sea bream is juvenile hermaphrodite and undif-ferentiated gonochoristic teleost. Male and female differentiation type of gonad is divided in undifferentiation stage, oogonia-like stage, ovary-like stage, ovary development stage, hermaphroditic testis stage, hermaphroditic ovary stage, and testis development stage. Undifferentiation stage is continued total lenth 18cm at 13 months after hatching. ovary-like stage is continued total length 11~18cm at 13 months after hatching. Ovary-like stage is continued total length 14~26cm at 10~14 months after hatching. Ovary development stage begins from total length 20cm, 14 months after hatching. At 20 months after hatching, 44 percent of total sampled individuals had ovary. Hermaphroditic ovary stage first begins total length 19~20 cm at 15 months after hatching, but it is not observed total length 28~29cm at 20months after hatching. Hermaphroditic testis stage first begins total length 21~22cm at 20months after hatching and is continued for 20months. Testis development stage first begins total length 20~21cm at 20 months after hatching, and is occupied 33 percent total length 28~29cm at 20 months. The beginning of sex differentiation more than 50 percent is from total length 16cm at 11 months after hatching. Sex determination begins total length 20cm, 14months after hatching in female and total length 20cm, 15 months after hatching in male. Sex determination more than 50 percent begins total length 23cm,, 17 months after hatching. Undifferentiated gonadal part of red sea bream consist gonad and fat body. As differentiation is going on and gonad is growing, fat body shrinks. This appearence is showed the same tendency in 3-year old red sea bream. 1.9mm larvae after hatching grow about 19mm larvae for 47 days. The relationship between the total length and body weight of larvae and juveniles in $BW=4.45{\times}10^{-6}TL^{3.4718}$ r=0.9820. Fishes in cage culture grow to maximum total length 28.4cm. The relationship between the total length and body weight of these fishes is $BW=2.36{\times}10^{-2}TL^{2.9180}$, r=0.9971. Undifferentiated gonadal part of red sea bream consist gonad and fat body. As differentiation is going on and gonad is growing, fat body shrinks.