• The Korean Journal of Malacology
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• v.31 no.4
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• pp.273-277
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• 2015

This study was conducted to provide the reproductive biological information and basic data on the artificial sex control of Haliotis discus hannai. The morphological sex differentiation process of H. discus hannai could be classified into following five phases: 1) formation of gonad outer membrane (FGOM) (${\leq}SL\;10.0{\pm}1.0mm$), 2) primordial germ cells (PGCs) appearance in the connective tissue between intestine and hepatopancreas (PAC), and formation of gonadal cavity (FGC) (SL $15.0{\pm}2.0mm$), 3) PGCs appearance in the epithelial layer of gonadal cavity (PAG) (SL $18.0{\pm}2.0mm$), 4) formation of gametogenic follicle and appearance of early oocytes and spermatogonia (FGOC) (SL $21.0{\pm}2.0mm$), 5) morphological sex differentiation (MSD) (${\geq}SL\;23.0{\pm}2.0mm$). From histological analysis sex differentiation rate in SL 24.1-25.0 mm of H. discus hannai was 90.0% and sex ratio (female : male) was 1:0.8.

• Journal of Aquaculture
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• v.9 no.4
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• pp.409-417
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• 1996

Age, annual growth pattern, and other ecologically important observations are reporting from the analysis of annual growth annuli of the sun and moon scallop, Amusium japonicum. Field survey made from 1993 to 1994 indicated that the sun and moon scallops are mostly distributed along the southern coast of Cheju-do, especially around the Sogwipo area. They inhabit on muddy sand bottom, at depth between 30 and 40 m where water temperature varies from 14 to $23^{\circ}C$ annually. The gonadal analysis indicated that the spawning mostly occurs during October and December and they exhibit multiple spawning peaks. The formation of annual growth annuli was found to be related with their spawning period. The shell growth on A. japonium japonicum appeared that their shell growth rate is comparatively faster than that of other scallops : 1. japonium japonicum reaches 6.28 cm in shell hight at the first year and 9.07 cm at the second year. The growth rate of gonadal somatic tissue was also found to be faster than growth rates of other scallops. One year old scallops weigh about 18.8 g, two and three year old scallops weigh 60 and 102.4 g. Based upon these observations, it is concluded that A. japonium japonicum can be a valuable species for aquaculture development due to their rapid growth.

• The Korean Journal of Malacology
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• v.20 no.1
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• pp.85-92
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• 2004

We investigated the reproductive cycle with gonadal development of the female Cyclina sinensis by histological observations and seasonal changes in biochemical components of the adductor muscle and visceral mass tissues were studied by biochemical analysis, from January to December, 2001. The reproductive cycle of this species can be classified into five successive stages: early active stage (February to April), late active stage (March to June), ripe stage (May to August), partially spawned stage (July to October) and spent/inactive stage (September to February). Total protein contents in the adductor muscle tissues reached the maximum in February (early active stage) and appeared the minimum in June (ripe stage), while their contents in the visceral mass tissues reached the maximum in the late active and ripe stages (June) and gradually decreased from July (partially spawned stage) to November (spent/inactive stage). Changes in total protein contents showed a negative correlationship between the adductor muscle and visceral mass tissues (r = -0.499, p = 0.099). Total lipid contents in the adductor muscle tissues reached the maximum in January (the inactive stages) and their contents gradually decreased from February. Their contents in the visceral mass tissues, however, reached the maximum in June (late active and ripe stage) and gradually decreased from July (the partially spawned stage). On the whole, total lipid contents showed a negative correlationship between the adductor muscle and visceral mass tissues (r = -0.631, p < 0.05). Therefore, These results indicate that the nutrient contents of the adductor muscle and visceral muscle tissues change in response to gonadal energy needs. Glycogen contents in the adductor muscle tissue reached the maximum in March (early and late active stages) and decreased from July to September (partially spawned stage). while their contents in the visceral mass tissues reached the maximum in June (late active and ripe stages) and gradually decreased from July (partially spawned stage). Thereafter, their levels gradually increased in November (spent/inactive stage). On the whole, changes in glycogen contents appeared negative correlationship between the adductor muscle and visceral mass tissues. However, they showed no significant different (r = -0.307, p = 0.331).

• Korean Journal of Fisheries and Aquatic Sciences
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• v.13 no.4
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• pp.135-144
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• 1980

The reproductive cycle and the breeding season of the freshwater clam, Anodonta (Sinanodonta) woodiana (Lea) have been investigated by histological examination of the gonadal development under photomicroscopy. The materials were monthly collected from the Nakdong River for one year from September 1979 to August 1980. Sexuality of Anodonta (Sinanodonta) woodiana is dioecious, and the species are ovoviviparous. The gonads are irregularly arranged from the subregion of mid-intestinal gland in visceral cavity to reticular connective tissue of foot. The ovary is composed of a number of small ovarian sacs, The epithelium of ovarian sac has a function of the germinal epithelium. Oogonia actively proliferate along the germinal epithelium of the ovarian sac, in which young oocytes are growing. The testis is composed of a number of seminiferous tubules, and the epithelium of the tubule has function of germinal epithelium, along which spermatogonia actively proliferate. A great number of undifferentiated mesenchymal tissue and eosinophilic granular cells are abundantly distributed between the growing oocytes and spermatocytes in the early development stages. With the further development of the ovary and testis these tissues and cells gradually disappear. Then the undifferentiated mesenchymal tissue and granular cells are considered to be related to the growing of the oocytes and spermatocytes. The gonads had function year-round the individuals which have various developmental stages of gonads appearing all the time. Spawning continued year-round except for the period of high temperature of water, during August and September. The peak spawning seasons appeared twice a year between January and March, and between June and July in 1980. Individuals which have trochophore larvae in the marsupium of the adult appeared year-round except September 1579 and August 1980. The rate of individuals which have glochidia in the marsupium was 72.7 percent in May 1980 which was the highest brooding fate.

• The Korean Journal of Malacology
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• v.27 no.1
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• pp.43-53
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• 2011

The gametogenic cycle and the spawning season in female and male Cyclina sinensis were investigated by quantitative statistical analysis using an image analyzer system, and the biological minimum size (the size at 50% of sexual maturity) was calculated by combination of quantitative data by size and von Bertalanffy's equation. Compared the gametogenic cycle by quantitative statistical analysis with the previous qualitative results in female and male C. sinensis, monthly changes in female and male gametogenic cycles calculated by quantitative statistical analysis showed similar patterns to the gonadal stages in female and male reproductive cycles by qualitative histological analysis. Comparisons of monthly changes in the portions (%) of each area to eight kinds of areas by quantitative statistical analysis in the gonads in female and male C. sinensis are as follows. Monthly changes in the portions (%) of the ovary areas to total tissue areas in females and also monthly changes in the portions of the testis areas to total tissue areas in males increased in March and reached the maximum in May, and then showed a rapid decrease from June to October. Monthly changes in the portions (%) of oocyte areas to ovarian tissue areas in females and also monthly changes in the portions of the areas of the spermatogenic stages to testis areas in males began to increase in March and reached the maximum in June in females and males, and then rapidly dropped from July to October in females and males when spawnig occurred. From these data, it is apparent that the number of spawning seasons in female and male C. sinensis occurred once per year, from July to October. Monthly changes in the number of the oocytes per mm2 and in the mean diameter of the oocyte in captured image which were calculated for each female slide showed a maximum in May and reached the minimum from December to February. Therefore, C. sinensis in both sexes showed a unimodal gametogenic cycle during the year. The percentage of sexual maturity of female and male clams ranging from 25.1 to 30.0 mm in length was over 50% and 100% for clams over 40.1 mm length. In this study, the biological minimum size (sexually mature shell lengths at 50% of sexual maturity) in females and males were 26.85 and 26.28 mm, respectively.

• Development and Reproduction
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• v.17 no.4
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• pp.469-475
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• 2013

In mammals, puberty is a process of acquiring reproductive competence, triggering by activation of hypothalamic kisspeptin (KiSS)-gonadotropin releasing hormone (GnRH) neuronal circuit. During peripubertal period, not only the external genitalia but the internal reproductive organs have to be matured in response to the hormonal signals from hypothalamic-pituitary-gonadal (H-P-G) axis. In the present study, we evaluated the maturation of male rat accessory sex organs during the peripubertal period using tissue weight measurement, histological analysis and RT-PCR assay. Male rats were sacrificed at 25, 30, 35, 40, 45, 50, and 70 postnatal days (PND). The rat accessory sex organs exhibited differential growth patterns compared to those of non-reproductive organs. The growth rate of the accessory sex organs were much higher than the those of non-reproductive organs. Also, the growth spurts occurred differentially even among the accessory sex organs; the order of prepubertal organ growth spurts is testis = epididymis > seminal vesicle = prostate. Histological study revealed that the presence of sperms in seminiferous tubules and epididymal ducts at day 50, indicating the puberty onset. The number of duct and the volume of duct in epididymis and prostate were inversely correlated during the experimental period. Our RT-PCR revealed that the levels of hypothalamic GnRH transcript were increased significantly on PND 40, suggesting the activation of hypothalamic GnRH pulse-generator before puberty onset. Studies on the peripubertal male accessory sex organs will provide useful references on the growth regulation mechanism which is differentially regulated during the period in androgen-sensitive organs. The detailed references will render easier development of endocrine disruption assay.

• Development and Reproduction
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• v.1 no.2
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• pp.189-202
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• 1997

The hypothalamic peptide GnRH plays a central role in the regulation of the mammalian reproductive axis. Recent studies suggested that GnRH stimulates or inhibits the ovarian steroidogenesis and gametogenesis directly. Our previous report indicated that GnRH gene is expressed in adult rat ovary as well as in hypothalamus and that the expressed GnRH may induce the follicular atresia and apoptosis of ovarian granulosa cells in rat. Therfore, we studied whether GnRH gene is expressed in the mouse fetal ovary, when the germ cells are degenerating by apoptosis during gonad diffeerentiation. Mouse fetal gonads were obtained on the 12, 15,18 and 20th day of gestation from the mother mice superovulated (10 IU PMSG and 10 IU hCG) and mated. The morphological changes of fetal ovaries were examined histochemically by hematoxylin-eosin staining. The fetal sex was confirmed by PCR methods for sexing. RT-PCR methods were used to examine the expression of GnRH gene and the sex steroid hormones were determined by conventional radioimmunoassays. The levels of estradiol (E) and progesterone (P) were increaseduntil 18th day of gestation and then E was decreased just before parturition. The morphological changes of fetal gonadal tissue sections showed the ovarian development and coincided with the result of PCR analysis for sexing using ovary- or testis- specific oligonucleotide primers. Immunoreactive GnRH in placenta was decreased gradually until the end of gestation but fetal brain and ovarian GnRH were increased. The level of GnRH gene expression was increased during fetal ovarian development from 12 till 18th day and decreased suddenly on 20th day just before birth. From these results, it is suggested that ovarian GnRh may play a regulatory role on the germ cell differentiation of fetal ovary.

• Journal of Embryo Transfer
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• v.28 no.2
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• pp.149-155
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• 2013

Mullerian inhibiting substance (MIS) is a member of the TGF-${\beta}$ (transforming growth factor-${\beta}$) family whose members play key roles in development, suppression of tumour growth, and feedback control of the pituitary-gonadal hormone axis. MIS is expressed in a highly tissue-specific manner in which it is restricted to male Sertoli cells and female granulose cells. The serum levels of MIS in prenatal and postnatal ICR mice were measured using the enzyme-linked immuno-solvent assay (ELISA) using the MIS/AMH antibody. Mice were grouped by age: the significant periods were at the onset of development. During sex organ differentiation, no remarkable difference between female and male foetus MIS serum levels (both<0.1 ng/ml) was observed. However, MIS serum levels in pregnant mice markedly changed (4.5~12.2 ng/ml). After birth, postnatal female and male mice serum MIS levels changed considerably (male: <0.1~138.5 ng/ml, female: 5.3~103.4 ng/ml), and the changing phase were diametrically opposed (male: decreasing, female: fluctuating). These findings suggest that MIS may have strong associations with not only develop-ment but also puberty. For further studies, establishing the standard MIS serum levels is of importance. Our study provides the basic information for the study of MIS interactions with reproductive organ disability, cancer, and the effect of other hormone or menopause. We hypothesise that if MIS is regularly injected into middle-age women, meno-pause will be delayed. We detected that serum MIS concentration curves change with age. The changing phase is different between males and females, and this difference is significant after birth. Moreover, MIS mRNA is expressed during the developmental period (prenatal) and also in the postnatal period. This finding indicates that MIS may play a significant role in the developmental stage and in growth after birth.

• Clinical and Experimental Pediatrics
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• v.54 no.5
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• pp.224-227
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• 2011

A sclerosing stromal tumor of the ovary is an extremely rare benign tumor; it usually is found during the second and third decades of life. Patients present with pelvic pain or a palpable abdominal mass. Hormonal effects such as masculinization are uncommon. Here, an 11-year old premenarchal girl presented with deepening of the voice. In addition, clitoromegaly and hirsutism with a male suprapubic hair pattern were observed. The laboratory findings showed that the testosterone level was elevated to 3.67 ng/ml, andostenedione to above 10 ng/ml, dehydroepiandrosterone-sulfate to 346 ${\mu}g$/dl and 17-hydroxy progesterone (17-OHP) to 11.28 ng/ml. The chromosome evaluation revealed a 46,XX female karyotype. An adrenocorticotropic hormone stimulation test was performed. The 17-OHP to cortisol ratio in 30 minutes was 0.045, which suggested a heterozygote for the 21-hydroxylase deficiency. However, the CYP21A2 gene encoding steroid 21-hydroxylase showed normal. The pelvic ultrasound showed a heterogeneous mass consisting of predominantly solid tissue in the pelvic cavity. The pelvic magnetic resonance imaging revealed an $8.9{\times}6.2{\times}6.6$ cm mass of the left ovary. A left oophrectomy was performed and microscopic examination confirmed a sclerosing stromal tumor. Immunohistochemical studies showed that the tumor was positive for smooth muscle actin and vimentin, but negative for S-100 protein and cytokeratin. Following surgery, the hormone levels returned to the normal range and the hirsutism resolved.

• Development and Reproduction
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• v.18 no.4
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• pp.301-309
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• 2014

Nesfatin-1, an anorexic nucleobindin-2 (NUCB2)-derived hypothalamic peptide, controls appetite and energy metabolism. Recent studies show that nesfatin-1/NUCB2 is expressed not only in the brain but also in gastric and adipose tissues. Thus, we investigated the distributions of nesfatin-1/NUCB2 in various tissues of male and female mice by real-time PCR, western blotting, and immunohistochemical staining. Real-time PCR analyses showed that NUCB2 mRNA was predominantly expressed in the pituitary and at lower levels in the hypothalamus, spleen, thymus, heart, liver, and muscle of both male and female mice. Expression was much higher in reproductive organs, such as the testis, epididymis, ovary, and uterus, than in the hypothalamus. Western blot analysis of the nesfatin-1 protein level showed similar results to the real-time PCR analyses in both male and female mice. These results suggest that nesfatin-1/NUCB2 have widespread physiological effects in endocrine and non-endocrine organs. In addition, immunohistochemical staining revealed that nesfatin-1 was localized in interstitial cells, including Leydig cells and in the columnar epithelium of the epididymis. Nesfatin-1 was also expressed in theca cells and interstitial cells in the ovary and in epithelial cells of the endometrium and uterine glands in the uterus. These results suggest that nesfatin-1 is a novel potent regulator of steroidogenesis and gonadal function in male and female reproductive organs. Further studies are required to elucidate the functions of nesfatin-1 in various organs of male and female mice.