• The Korean Journal of Malacology
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• v.22 no.2
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• pp.125-134
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• 2006

We investigated the reproductive cycle of the hard clam, Meretrix petechialis with its gonadal development by histological observations. The seasonal changes in biochemical component of the adductor muscle, visceral mass, foot muscle and mantle of the clam were studied by biochemical analysis, from January to December, 2002. The reproductive cycle of this species can be divided into five successive stages: early stage (January to March), late active stage (February to May), ripe stage (April to August), partially spawned stage (July to August) and spent/inactive stage (September to January). Total protein content in the visceral mass was over two times higher than that in the adductor muscle. Monthly changes of total protein content in the adductor muscle were not statistically significant (ANOVA, p = 0.071), while the changes in the visceral mass were significant (p < 0.001). Total protein content in visceral mass was higher during the early active, late active, and ripe stages (from January to May), while the lowest in July. Glycogen content in the adductor muscle was higher than that in the visceral mass. Monthly changes in glycogen contents were statistically significant in both adductor muscle (F = 237.2, p < 0.001) and the visceral mass (F = 64.04, p < 0.001). Glycogen content in the adductor muscle was the highest in the ripe stage (April). Its content was lower in the partially spawned and the spent/inactive stages (June-September). Glycogen contents in the visceral mass were relatively lower until the early active stage, while the highest in the late active stage. RNA content was higher in visceral mass than that in the adductor muscle. Monthly changes in RNA contents were significant in both adductor muscle (F = 195.2, p < 0.001) and visceral mass (F = 78.85, p < 0.001). RNA content in the adductor muscle was high in the early active stage (January-February), and then it decreased rapidly in the late active stage (March-April), thereafter, slightly increased during the partially spawned stage (June-July). RNA content in the visceral mass reached a maximum during the ripe stage (May), and then it decreased rapidly during the partially-spawned stage (June-July). There was significant positive correlation in total protein contents between adductor muscle and visceral mass (r = 0.715, p = 0.020). However, there was no correlation between adductor muscle and visceral mass in glycogen (p = 0.550), while a negative correlation was found between the adductor muscle and visceral mass in RNA (p = 0.518) contents. Especially, changes in RNA content showed a negative correlation between the adductor muscle tissue and visceral mass. Therefore, these results suggest that the nutrient content of the adductor muscle, visceral muscle and foot muscle changed in response to gonadal energy needs.

• Journal of Aquaculture
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• v.11 no.4
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• pp.475-485
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• 1998

To clarify annual reproductive cycle of Koran dark sleeper, odontobutis platycephala, we examined the seasonal changes of gonadosomatic index(GSI), testicular development stages and sex steroid hormones in blood from December 1995 to November 1997. Testis was podlike shape from July to October, and tadpole-like shape from November because of its expanded posterior part. GSI was 0.14~0.18 from July to September and increased to $0.43{\pm}0.04$ in October and then was not changed significantly until February. GSI was reincreased to $0.52{\pm}0.09$ from March and then was kept at similer levels until May, but fell down to $0.28{\pm}0.05$ in June. As results of histological observation, testis was divided into 3 parts(anterior, boundary, posterior) in the development progress of germ cells. In July, the testis was composed of only spermatogonia without seminiferous tubules in most fishes. In the anterior part of testis, the ferquency of spermatogenesis stage seminiferous tubules appearing in August was more than 80% from September to December. decreased gradually from January to March and drastically in April, and then disappeared in June. The frequency of spermiogenesis stage seminiferous tubules appearing in December, increased gradually from January to March and drastically to 80% in April, and reached to 90% the highest levels of the year in June. Post-spawning stage seminiferous tubules did not appear throughout the year. The frequency of spermatogonia was 100% and 65% in July and August, and less than 20% in the rest period of the year. In the boundary part, the frequency of spermatogenesis stage seminiferous tubules appearing in August increased from September and reached to 82% in November, decreased from December, adn disappeared in March. The frequency of spermiogenesis stage seminiferous tubules appearing in November was less than 18% until February, and increased to 29%~57% from March to June. The frequency of post-spawning stage seminiferous tubules appeared 12%~25% only from March to June. The frequency of spermatogonia was 100% in July, decreased to 85% in August and 10% in November, and increased gradually from December to 50% in April, and decreased again from May to June. In the posterior part, seminiferous tubules with some seminiferous tubules increased drastically 80%~85% in August and September, decreased drastically from October to November and remained below 10% until February, and disappeared after March. The frequency of spermiogenesis stage seminiferous tubules appearing in August increased sharply from October and reached to 75% in November. decreased to 15% in December and no significant changes until March, and disappeared after April. The frequency of post-spawning stage seminiferous tubules appearing very early in November increased to 82% in December and 85%~95% until June. The frequency of spermatogonia was 100% in July, decreased drastically to 15% in August, disappeared from October to Mrch, but reappeared from April and kept at less than 10% until June. The blood level of testosterone (T) increrased gradually from August was $0.61{\pm}0.09 ng/m\ell$ in November, increrased drastically to $3.99{\pm}1.22 ng/m\ell$ in December and maintained at in similar level until March, and decreased to $0.25{\pm}0.14 ng/m{\ell} ~ 0.17{\pm}0.13ng/m{\ell}$ in April and May and no significant changes until July (P<0.05). The blood level of 17, 20 -dihydroxy-4-pregnen-3-one $ng/m{\ell}$in the rest of year without significant changes(P<0.05). Taken together these results, the germ cell development of testis progressed in the order of posterior, boundary, anterior part during annual reproductive cycle in Korean dark sleeper. The testicular cycle of Korean dark sleeper was as follows. The anterior part of testis : i.e. spermatogonial proliferation period (July), early maturation period (from August to November), mid maturation period (from December to March), late maturation period (from April to May) and functional maturation period (June) were elucidated. The boundary of testis, i.e. spermatogonial proliferation period (July), early maturation period (from August to October), mid maturation period (from November to February) and the coexistence period of late maturation, functional maturation and post-spawn (from March to June) were elucidated. The posterior of testis, i.e. spermatogonial proliferation period (July), mid maturation period (from August ot September), late maturation period (October), functional maturation period (November) and post-spawn period (from December to June) were elucidated. It was showed that the changes of sex steroid hormone in blood played a important roles in the annual reproductive cycle of Korean dark sleeper.

• Development and Reproduction
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• v.9 no.2
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• pp.95-103
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• 2005

Reproductive cycle, gonadosomatic index(GSI), egg diameter composition, first sexual maturity, sexually matured length(50% of first sexual maturity), and sex ratio of Amusium japonicum japonicum, were investigated by histological observations and morphometric data. Samples were collected monthly from the subtidal zone of Sogwipo, Jejudo, Korea, for two years. The sun and moon scallop Amusium japonicum japonicum is dioecious. Monthly variation in the GSI showed similar patterns with the reproductive cycle. Ripe oocytes were about $70{\sim}90\;{\mu}m$ in diameter and had thick egg membranes. The spawning period was from November to January, and the main spawning occurred between November and December when the seawater temperature was relatively low. From monthly changes in egg diameter composition, the spawning period was once a year, although the number of spawning frequencies is assumed to occur more than twice during the spawning season. The reproductive cycle of this species could be divided into five successive stages: early active stage(April to June), late active stage(June to September), ripe stage(October to November), spawning stage(November to January), and spent/resting stage(February to April). First sexual maturities in female and male scallops ranging from 85.1 to 90.0mm in shell length were over 50% and they were 100% for scallops over 90.0mm in shell length. In this population, sexually matured shell lengths(50% of rate of group maturity) in females and males were 86.96 and 86.59mm, respectively. The female to male sex ratio among individuals over 85.1mm in shell length was not significantly different from 1:1($X^2=0.18$, p>0.05). No evidence of hermaphrodite was found in histological sections of any scallop examined.

• The Korean Journal of Malacology
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• v.21 no.2 s.34
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• pp.121-132
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• 2005

We investigated the reproductive cycle with gonad developmental phases of Sinonovacula constricta by histological observations, and seasonal changes in biochemical component of the adductor muscle, visceral mass, foot muscle and mantle were studied by biochemical analysis, from January to December, 2004. The reproductive cycle of this species can be classifed into five successive stages: early active stage (March to May), late active stage (May to July), ripe stage (July to September), partially spawned stage (August to October) and spentfinactive stage (October to March). Total protein content was the highest in the following order: adductor muscle, visceral mass, foot muscle, and mantle. Except for mantle, it was generally higher during the ripe and spawning stages, while lower during the spent/inactive stage. There were positive correlations in total protein contents among adductor muscle, foot muscle, and visceral mass. However, the correlations were not statistically significant. Total lipid content was the highest in the visceral mass; it was more than 5 or 6-fold higher than those in the adductor muscle, foot muscle, or mantle. The monthly change was also most dynamic in the visceral mass. It first Increased during the early active stage (March to May), decreased during late active stage (May to July), and then increased again rapidly during the spawning stage (September). There were a strong negative correlation in total lipid contents between foot muscle and adductor muscle (r = -0.634, p = 0.027), and a strong positive correlation between adductor muscle and mantle (r = 0.665, p = 0.018). Glycogen contents showed more or less similar pattern to total lipid contents in the adductor muscle, foot muscle, and visceral mass. It was higher during the early active and spawning stages, while lower during the late active and spent/inactive stages. There was no statistically significant correlation in glycogen contents among different tissues. Especially, total lipid content showed a negative correlationship between the foot muscle, adductor muscle, visceral mass and mantle. Therefore, these results indicate that the nutrient content of the foot muscle, adductor muscle, viseral mass and mantle changed in response to gonadal energy needs.

• Development and Reproduction
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• v.11 no.3
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• pp.155-165
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• 2007

Gonad index (GI), conditon index, the reproductive cycle with gonadal development of the Neptunea (Barbitonia) arthritica cumingii, were investigated histologically, based on the samples which have been collected from the subtidal zone of Oeyeondo, Boryeong, Korea from January to December, 2006. Neptunea (Barbitonia) arthritica cumingii is dioecious and oviparous. Monthly changes in the gonad index (GI), studied for determination of spawning period, were closely associated with gonad developmental phases. The GI reached a maximum in April, and gradually decreased from May to August due to spawning. The gonadsomatic index and condition index showed similar patterns to gonad developmental phases and the spawning period. The reproductive cycle according to gonad developmental phases of this species can be classified into five successive stages in females and males: in females, early active stage (September to October), late active stage (November to February), ripe stage (February to June), partially spawned stage (May to August) and recovery stage (June to August); in males, the early active stage (September to October), late active stage (November to February), ripe stage (February to June), copulation (April to July), and recovery stage (July to August). Spawning occurred between May to August in females and April to July in males, and spawning peak in females was observed between June and July when the seawater temperature rose to above $19^{\circ}C$. Percentages of first sexual maturity of female and male snails ranging from $50.1{\sim}60.0\;mm$ in shell height were over 50%, and 100% for snails over 60.1 mm in shell height. The sex ratios of females to males were not significantly different from a 1:1 sex ratio.

• Clinical and Experimental Reproductive Medicine
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• v.30 no.1
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• pp.95-103
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• 2003

Objective : To assess and compare the clinical outcomes between GnRH agonist long protocol and GnRH antagonist short protocol in oocyte donation program. Materials and Methods: Of total 18 oocyte donation cycles, controlled ovarian hyperstimulation (COH) were performed with GnRH agonist long protocol and GnRH antagonist short protocol in initial 9 cycles and later 9 cycles, respectively. Oral estradiol valerate and progesterone in oil we re administrated to all recipients for endometrial preparation. Oral estradiol administration was started from donor cycle day 1 after full shut down of gonadal axis with GnRH agonist in patients with ovarian function. Progesterone was injected from oocyte retrieval day of donor initially, then continuously till pregnancy 12 weeks if pregnancy was ongoing. We compared the parameters of clinical outcomes, such as number of the retrieved oocytes, fertilization rate, high grade embryo production rate, clinical pregnancy rate, implantation rate, ongoing pregnancy rate, COH duration, total gonadotropin dose for COH between GnRH agonist long protocol group and GnRH antagonist group. Statistical analysis was performed using Mann-Whitney test, p<0.05 was considered as statistically significant. Results: The number of retrieved oocytes, fertilization rate, high grade embryo production rate, clinical pregnancy rate, implantation rate, ongoing pregnancy rate were $14.89{\pm}7.83$, 81%, 64%, 78%, 31%, 78%, respectively in GnRHa long protocol group and $11.22{\pm}8.50$, 79%, 64%, 67%, 34%, 56%, respectively in GnRH antagonist group. There was no significant differences in parameters of clinical outcomes between 2 groups (all p value >0.05). Duration and total gonadotropin dose for COH were $10.94{\pm}1.70$ days and $43.78{\pm}6.8$ vials in 18 cycles, $12.00{\pm}1.73$ days and $48.00{\pm}6.93$ vials in agonist group, $9.88{\pm}0.78$ days and $39.55{\pm}3.13$ vials in antagonist group, respectively. In GnRH agonist long protocol group, significantly longer duration and higher gonadotropin dose for COH were needed (p=0.012). Conclusion: In oocyte donation program, clinical outcomes from controlled ovarian hyperstimulation with GnRH antagonist were comparable to those from GnRH agonist long protocol group, so controlled ovarian hyperstimulation with GnRH antagonist may be effective as GnRH agonist long protocol. At least there may not be harmful effects of GnRH antagonist on oocyte development and quality.

• Korean Journal of Ichthyology
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• v.11 no.2
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• pp.117-125
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• 1999

We investigated the reproductive cycles of two freshwater fishes, Moroco oxycephalus and M. lagowskii, in Korea. Seasonal changes in gonadosomatic index (GSI) and gonads were investigated histologically from April 1998 to April 1999. The reproductive cycles of two species were not shown any differences. The reproductive cycle can be divided into 5 phases : phase I (spent phase), phase II (immature phase), phase III (early developing phase), phase IV (late developing phase), and phase V (ripe phase). In phase I, the gonads of two species began to lose distinctly their weights from mid April, and reached the lowest GSI in late July (phase II). In September, the GSI values of testis and ovary increased very slowly (phase III) and gonadal developments rested during the winter season (phase IV). In March, the GSI values of M. oxycephalus and M. lagowskii began to increase, and reached the maximum in April (phase V). From the cyclic changes in the GSI and histological analyses, the spawning period was between mid April and mid May.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.19 no.1
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• pp.83-91
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• 1986

The activity of the liver cells of greenling, Agrammus agrammus were histologically investigated under photo-and electron microscopy, and studied by comparing seasonal changes of hepatosomatic index (HSI). The materials were monthly collected at the costal area of Tongbaeksom, Pusan, Korea, from September 1983 to August 1984. The annual variations of HSI of male were not distinct, but those of HSI in female began to increase in autumn, and reached the maximum in winter when the ovary was getting mature. During the period of yolk accumulation in the oocytes, the female liver and its hepatic cells were seen to large and nuclei and nucleoli were hypertrophic also. At this time the amounts of glycogen and lipid in the cells gradually decreased, while basophilic substance (RNA) increased. And well-developed granular endoplasmic reticula binding ribosomes were supposed to play the leading role in protein synthesis and deposition for vitellogenin in the cystoplasm. Just prior to spawning, glycogen and lipid droplets were decreased, but basophilic substances(RNA) were found in a high concentration especially at the peripheral region of the liver cells of females. In the liver cells of males, were hardly altered by gonadal maturation, basophilic substances gradually increased, glycogen particles and lipid droplets were still observed in large quantities. After spawning, basophilic subtances decreased in the liver cells of female and male.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.24 no.3
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• pp.167-176
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• 1991

Sexual maturation of the bluespotted mud hopper, Boleophthalmus Pectinirostris(Linnaeus) was investigated histologically on the gonadal development, and studied by gonadosomatic index(GSI), egg diameter composition. Samples were collected in the intertidal zone of Wolyon-ri, Hoihyon-myon, Okku-gun, Chollabuk-do, Korea, from April to October in 1988 and from June to August in 1989. The ovary is a pair of sac-shaped organ. The testis is a pair of tubule-shaped organ and it is connected to the seminal vesicle which is located at the posterior end of the testis. In male and female, GSI began to increase from late May when the water temperature began to increase and reached the maximum value in June and July, respectively. It began to decrease from August, the highest water temperature season. Thereafter, maintained relatively low values until October. The annual reproductive cycle of this species could be classified into four sucessive developmental stages: growing stage$(April{\~}May)$, mature stage$(June{\~}early\;July)$, ripe and spent stage(late lune-early August), degenerative and resting stage$(late\;August{\~}March:\;the wintering\;period)$. According to the frequency distributions of egg diameters in the spawning season, Boleophthalmus Pectinirostris was species to spawn twice or more in the spawning season.

• Journal of Aquaculture
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• v.21 no.4
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• pp.331-338
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• 2008

Monthly changes in the gonadosomatic index (GSI) and hepatosomatic index (HSI) of wild river puffer Takifugu obscurus, and water quality environment in spawning area during breeding season were investigated from March 1995 to February 1996. Monthly changes in GSI and HSI of T. obscurus, that was cultured in low salinity, were calculated. The external morphology of the gonads, germ cell differentiation during gametogenesis and the reproductive cycle with the gonad developmental phases were investigated by histological analysis. The optimum water quality environment in Ganggyung, Choongcheongnam-do, where is spawning ground of wild T. obscurus, was $15-20^{\circ}C$ (water temperature) and 0 psu (salinity). Monthly changes in the GSI in females and males reached a maximum in May, and then rapidly decreased. Therefore, it is assumed that in the natural condition the spawning period of wild T. obscurus is May to June. In females and males, it showed a negative correlationship between the GSI and HSI. The external morphology of the gonads in female and male T. obscurus, that was cultured in low salinity, is composed of a pair of saccular structure. Based on monthly changes in the GSI, it is assumed that in female T. obscurus, that was cultured in low salinity, spawn from March through May. Therefore, it showed a negative correlationship between changes in the GSI and HSI. On the whole, in females and males, it showed a similar pattern between wild and cultured T. obscurus. The reproductive cycle with the gonad developmental phases can be classified into successive five stages in females: the early growing stage, late growing stage, mature stage, ripe and spent stage, and recovery and resting stage. In males, that can be divided into successive four stages: the growing stage, mature stage, ripe and spent stage, and recovery and resting stage. In case of wild T. obscurus, the spawning period has once a year, however, those cultured in the high water temperature ($20-27^{\circ}C$) - low salinity (under 3.3 psu) condition have reproductive characteristics having possibilities of discharge of eggs and sperms year-round as a multiple spawner.