• Development and Reproduction
• /
• v.4 no.2
• /
• pp.187-193
• /
• 2000

Gonad and blood samples were taken from the cultured female Korean sea bass, Lateolabrax japonicus from October to February between 1997 and 1999. Gonadosomatic index began to increase in November and reached the highest value in December (12.8$\pm$1.5) and January (14.8$\pm$3.5), and then decreased sharply in February (2.6$\pm$1.5, p<0.05). The ovarian oocytes developed to tertiary yolk stage and reached fully-Brown stage in December and January, and then underwent atresia without maturation and ovulation in February. The plasma estradio3-17 $\beta$ level increased from November, and reached the highest value in December (1,152.3$\pm$107.2 pg/ml) and January (1,315.4$\pm$99.5 pg/ml), after then decreased in February (P<0.05). The concentration of plasma 17 $\alpha$ ,20 $\beta$-dihydroxy-4-pregnen-3-one was not significantly changed at low levels (86.6$\pm$6.5∼93.8$\pm$2.8 pg/ml) during the experimental period (P<0.05). All the fish with fully-grown oocytes in the ovary were matured and ovulated by HCG injection. The number of floating eggs were 325,000$\pm$26,000 at HCG 1,000 luhg and 195,000$\pm$35,000 at 2,000 lUikg. There was no difference in fertilization rate and hatching rate of the eggs (P<0.05). Considering these results, we could infer that the ovarian oocyte of the cultured Korean sea bass were not matured and ovulated because of the lack of gonadotropin surge. Moreover, HCG injection could induce oocyte maturation and ovulation in the cultured fish, and the effective dose was 1,000 IU/kg.

• Development and Reproduction
• /
• v.8 no.1
• /
• pp.57-64
• /
• 2004

This study was carried out to examine the expression of the circadian clock genes in the mouse ovary and testis at different developmental stages. Expression of Period1(Per 1), Period2(Per2), Period3(Per3), Cryptochrome1(Cry1), Cyptochrome2(Cry2), Clock Small and Prokineticin1 and Prokineticin2 receptor(Prok1r, Prok2r) genes in mouse ovary was explored by semiquantitative reverse transcription Polymerase chain reaction(RT-PCR) according to the developmental stage(post partum day; ppd 1, 7, 10, 21 and 35). Immunohistochemistry using PER1 antibody was also analyzed. The differential expression pattern of clock genes was presented according to stages of the mouse ovarian development (ppd 1, 7, 10, 21 and 35). In the cases of ovaries, at the starting point of follicle growth at ppd 7 and 10, the clock gene expression patterns were changed vastly. According to the developmental stages, the clock genes were highly expressed at ppd 7 and 10 in mouse testis also. Receptors for Prok2, the circadian output molecule of SCN, were also expressed in ovary at ppd 7 and in testis at ppd 1 and 7, respectively. Immnunohistochemical analysis of PER1 showed positive signals in the cytoplasm of oocytes and granulosa cells. The level or PER1 expression was increased in cells at the spermatogonia and the condensing spermatids. The expression pattern of Perl and localization of PER1 were showed similar patterns according to the developmental stages in ovary and testis. Taken together, it could be observed that the expression of clock genes was highly correlated with gonadal development and germ cell differentiation in mice. Therefore, in this study, circadian programming of the genes in the ovary and testis is strongly imposed across a wide range of core reproductive cycles and normal development of gametes. Although the existence of circadian genes is clearly investigated, further studies on the direct evidence is required for the understanding of the relationship between circadian genes and regulation of gonadal differentiation and germ cell development.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.13 no.4
• /
• pp.125-134
• /
• 1980

The dovelopment of the gonads, gametogenesis and the reproductive cycle of the topshell, Turbo cornutus Solander, which is one of valuable food animals fom Korean waters were studied by photomicroscophy. The materials were monthly collected from Bangeojin, Jeongjari and Dangweol, all these places being located in the south-eastern part of Korea, for one year from March 1979 to February 1980. Topshell is dioecious and oviparous. Gonad is situated on the surface of liver, which lies posteriorly. The surface of ovary and testis is covered with a fibrous membrane, membrane of connective and muscular fibers and then an outermost layer of simple-columnar epithelial cells which are composed of cuboidal and columnar mucous gland cells. Primordial germ cells develop on the germinal epithelium of ovarian and testicular lobuli which are originated from the fibrous membrane and extend toward hepatic gland. Undifferentiated mesenchymal tissue and pigment 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 tissue and cells gradually disappear. Then the undifferentiated mesenchymal tissue and pigment granular cells are considered to be related to the growing of the oocytes and spermatocytes. Early multiplicating oogonium is ca. $10\mu$ in diameter and nucleushaving a central nucleolus is ra. $8\mu$. As the oocytea grow to ca. $50-60\mu$ by the increase of cytoplasm, the oocytes become look like bunches of grapes which are attached to ovarian lobuli. Mature eggs are ca. $180-210\mu$ in diameter and it is surrounded by a gelatinous membrane of ca. $10\mu$ in thickness. After spawning, undischarged ripe eggs and spermatozoa remain in the ovary and testis respectively for some time. Then they finally degenerate, and proliferation of new oogonia and spermatogonia occur along the germinal epithelia of newly developed ovarian and testicular lobuli. Reprocuctive cycle of Turbo cornutus could be classified into five successive stages: multiplicative, growing, maturer spent and recovery stages. Spawning occurs from August to November with Peak spawning from early September to late October.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.17 no.3
• /
• pp.206-218
• /
• 1984

The gonadal development and gametogenesis of shad, Konosirus, punctatus (TEMMINCK et SCHLEGEL) were studied by comparing with various quantitative indices, such as seasonal changes of gonadosomatic index, fatness, egg-diameter composition, first maturing size, and by comparing with histological changes of gonad and gonadotrophs(GTH) in pituitary. The materials were monthly sampled from Dadaepo at the estuary of the Nakdong river in Korea from September, 1982 to October, 1983. The ovary of shad is a pair of sac-shaped organs revered with a fibromuscular capsule and consisting of numerous sacs. The type of testicular structure is lobular type with development of germ cells, mesenchymal tissue on the lobuli. The gonadosomatic index (GSI) is rather low till March, but increases in April and reaches to peak in June in females and May in males. And it suddenly falls in July. The gonads become active on the increase of water temperature and spawning season ends before high water temperature. After spawning, the small oocytes continue to remain as they are untill the growing period next year. The reproductive cycle includes the successive stages of growing from March to April, mature from April to May, ripe and spawning in June, and recovery and resting from July to February next year. In egg-diameter composition of an ovary taken in the spawning season, 2-3 modes were recognized with some batches shown in an ovary. An individual shad spawns twice or more in a month-spawning season. The individual spawning interval is estimated to be ten days or less. Changes of fatness are corelated with those of water temperature that affect on the condition of feeding, but less corelated with spawning. The percentage of mature of female and male fish, are $50\%$ in 17.0-18.0 cm and $100\%$ in 18.0-19.0 cm. GTH cells are activated from growing period and decrease their activity at pre-spawning season with peak activity for mature period.

• Korean Journal of Ichthyology
• /
• v.19 no.1
• /
• pp.8-15
• /
• 2007

The morphology of gonad and reproductive cycle of ribbed gunnel (Dictyosoma burgeri) were investigated on the basis of histological observation. The specimens were monthly sampled in the coastal waters of Jeju from November 2001 to February 2003. The ovaries and testis of this species are categorized as cystovarian and lobule type, respectively. The gonadosomatic index (GSI) of female increased in November and maintained high values from December to February. The GSI of male was similar to that of female although it was decreased in February. The reproductive cycle can be grouped into the following successive stage in the ovary: growth (October to November), mature (November to February), spawning (January to February), and degenerating and recovery (March to September). And in the testis, the stage observed were: multiplication (August to November), growth (November to January), mature and spawning (November to February), and degenerating and recovery (January to September). The minimum maturation size of D. burgeri was over 15.0 cm and fecundity ranged from 2,194 to 6,581 eggs. The relationship between the fecundity and fish body was calculated in the fecundity (F) equation as: $F=0.4057TL^{3.1425}$ ($R^2=0.7621$) for total length (TL); $F=149.88BW^{0.9579}$ ($R^2=0.7982$) for body weight (BW), respectively. The fecundity was correlated positively with TL and BW. The histological observations of the gonads suggested that major spawning of this species probably occurs between January to February, when low water temperature ($13{\pm}0.3^{\circ}C$) period.

• Journal of Aquaculture
• /
• v.11 no.2
• /
• pp.173-182
• /
• 1998

The study has been conducted to understand gonadal development and the effects of $17^{\alpha}$-methyltestosterone on sex inversion of the red spotted grouper, Epinephelus akaara. Fish were collected from Deukyand bay in the southern coast of Korea in August, 1996 and then they had been cultivated at the indoor tank until August, 1997. Gonad somatic index (GSI) in the females of both treated and control group began to increase from February when water temperature was rainse again, and reached the maximum value in August, whereas it had decreased from September adn thereafter maintained relatively low value until January. Unlike females, GSI in the male or intersex of treated groups decreased after June. Hepatosomatic index (HSI) of the control group tended to show the relatively low around Autumn, whereas it showed relatively highr value in April and June when the ovary was in the growing stage. Although the treated groups showed relatively higher value of the HSI than the control, hte paterns in monthly variation of HSI were similar to the control. Sexual change of the female grouper to the male was attempted by acceleration with oral administration of $17^{\alpha}$-methyltestosterone at the dose of 0.2 and 0.5mg/kg fish for 120days. Transitional hermaphroditic gonads were observed from the various size of groupers ranging 21.0 to 36.1 cm in total length, while the functional males could be induced from th individuals of 28.8 to 33.5cm in total length. This result indicated that larger groupers than 30cm in total length should be used for sex inversion to maleness with $17^{\alpha}$-methyltestosterone.

• Korean Journal of Poultry Science
• /
• v.28 no.2
• /
• pp.135-142
• /
• 2001

A novel sterategy has been established to determine the origin of the Primordial Germ Cells (PGCs) in avian embryos directly and the developmental fate of the PGCs for the application to Poultry biotechnology. Cells were removed from 1) the centre of area pellucida, 2) the outer of area pellucida and 3) the area opaca of the stage X blastoderm (Eyal-Giladi & Kochav, 1976). When the cells were removed from the centre of area pellucida, the mean number of circulating PGCs in blood was significantly decreased in the embryo at stage 15 (Hamburger & Hamilton, 1951) as compared to intact embryos. When the cells were replenished with donor cells, no reduction in the PGCs number was observed. The removal of cells at the outer of area pellucida or at the area opaca had no effect on the number of PGCs. In case, another set of the manipulated embryos were cultured ex vivo to the hatching and reared to the sexual maturity, the absence of germ cells and degeneration of seminiferous tubules was observed in resulting chickens derived from the blastoderm in which the cells were removed from the centre of the area pellucida. It was concluded that the avian Primordial Germ cells are originated at the center of area pellucida. Developmental ability of the cells to differentiate into somatic cells and germ cells in chimeras were analyzed. Somatic chimerism was detected as black feather attributed from donor cells. Molecular identification by use of female - specific DNA was performed. It was confirmed that the donor cells could be differentiated into chimeric body and erythrocytes. Donor cells retained the ability to differentiate into germline in chimeric gonads. More than 70% of the generated chimeras transmitted donor derived gametes to their offspring indicating that the cells at the center of area pellucida had the high ability to differentiate into germ cells. A molecular technique to identify germline chimerism has been developed by use of gene scan analysis. Strain specific DNA fragments were amplified by the method. It would be greatly contributed for the detection of germline chimerism. Mixed- sex chimeras which contained both male and female cells were produced to investigate the developmental fate of male and female cells in ovary and testes. The sex combinations of donor and recipient of the resulting chimeras were following 4 pairs; (1) chimeras (ZZ/ZZ) produced by a male donor (ZZ) and a male recipient (ZZ), (2) chimeras (ZW/ZW) produced by a female donor (ZW) and a female recipient (ZW), (3) chimeras (ZZ/ZW) Produce by a male donor (ZZ) and a female recipient (ZW), (4) chimeras (ZW/ZZ) produced by a female donor (ZW) and a male recipient (ZZ). It was found that genetically male avian germ cells could differentiate into functional ova and that genetically female germ cells can differentiate into functional spermatozoa in the gonad of the mixed- sex chimeras. An ability for introduction of exogenous DNA into the PGCs from stage X blastoderms were analyzed. Two reporter genes, SV-$\beta$gal and RSV-GFP, were introduced into the PGCs. Expression of bacterial/gal was improved by complexing DNA with liposome detectedcc in 75% of embryos at 3 days embryos. At the embryos incubated for 1 day, expression of the GFP was observed all the embryos. At day 3 of incubation, GFP was detected in about 70% of the manipulated embryos. In case of GFP, expression of the transgene was detected in 30 %e of the manipulated embryos. These results suggested that the cells is one of the most promising vectors for transgenesis. The established strategy should be very powerfull for application to poultry biotechnology.

• The Korean Journal of Malacology
• /
• v.24 no.2
• /
• pp.153-159
• /
• 2008

We investigated artificial mass seed production of a Chinese scallop, Patinopecten yessoensis, in 2004. The GSI(gonad somatic index) of the Chinese scallop, P yessoensis was 17.2 on mid-February, 20.2 on mid-March, while that of Korean scallop, P yessoensis was 6.9 on mid-February, 10.8 on mid-March. Matured 120 females and 350 males were selected for artificial mass production. They were exposed in air for 1 hr at over $20^{\circ}C$, and placed into a spawning tank(20 ton) containing sea water treated with UV radiation at $12^{\circ}C$. We gained a total of 228,000 thousand scallop embryos between March 10th and 15th, and reared larvae at the indoor tank during 25 days. When the mean shell length of larvae reached 250 ${\mu}m$ and they have eye-spots, the number of pre-settling larvae was 47,500 thousand. We gained 1,850 thousand attached scallop spats from two kinds of collectors. Attached spats were reared in indoor tank for different periods from 5 days to 60 days. They were divided into 5 groups according to the length of reared days. Each group of attached spats was moved to intermediate rearing sites at Yangyang fishing port in Gangreung-city for acclimation to ocean environments. The highest survival rate of attached spats was 13.0% shown at the group reared for 12 days, but the significant difference in their growth was not found between the groups. The shell length of artificial attached spats increased from 0.9 ${\mu}m$ on July 10th to 24.7 ${\mu}m$ on December 16th with the survival rate of 85.0% while that of natural attached spats increased from 0.6 ${\mu}m$ on July 10th to 23.9 ${\mu}m$ on December 16th with the survival rate of 85.7%.

• Korean Journal of Ichthyology
• /
• v.19 no.2
• /
• pp.107-111
• /
• 2007

Sex differentiation of the brown croaker Miichthys miiuy (Basilewsky) is described from hatching to the 120th day post-hatching (dph) (water temperature $24^{\circ}C$). Primordial germ cells (PGCs) were observed on the 20th dph (10.4 mm total length (TL), 0.14 g body weight (BW), and began to protrude into the peritoneal cavity from the 40th dph (19.4 mm TL, 0.39 g BW). On the 65th dph (31.3 mm TL, 0.93 g BW, $1,560D^{\circ}$ (degree-days)), initial ovarian differentiation was identified by the PGCs with condensed chromatin, and their transformation into meiotic oocytes. By the 120th dph (4.60 mm TL, 1.38 g BW, $2,880D^{\circ}$), the oocytes were in the perinucleolus stage and had increased from 20 to $40{\mu}m$ in diameter. While ovaries gradually grew after sex was differentiated, testes continued to multiply from the 65th dph. On the 80th dph (37.9 mm TL, 1.39 g BW, $1,920D^{\circ}$), the beginning of testis lobule formation was indicated by the occurrence of spermatogonial cysts enveloped by somatic cells in some of the testes. On the 120th dph, the testis lobules of some of the fish contained all germ cell stages through to the spermatocytes. Therefore, the sex differentiation type of the brown croaker is identified as gonochoristic.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.15 no.3
• /
• pp.241-250
• /
• 1982

The structure of gonads, gametogenesis and reproductive cycle of the cockle, Fulvia mutice, were studied mainly by histological observation. The materials were monthly sampled in the southern area of Yeosu from October 1980 to September 1981. F. mutica was monoecious. The gonads were situated between the liver tissues and the outer fibronuscular layers compacted by the connective tissue fibers and muscle fibers beneath the outermost layer of simple cuboidal epithelium. The gonad was composed of a number of the ovarian sacs and the testicular tubules which form the tubular structure. Testicular tubules in the mature stage sometimes contained 'testis-ova' The undifferentiated mesenchymal tissues and the eosinophilic cells were abundantly distributed on the germinal epithelium in the early development stage. With the further development of the ovary and testis, these tissues and cells gradually disapprared. The undifferentiated mesenchymal tissues and the eosinophilic cells are related to the growing of the oocytes and spermatocytes . Early multiplicating oogonium was about $10{\mu}m$ in diameter. As the oocytes grow to $27-34\times50-58{\mu}m$ by increasing cytoplasm, the oocytes connected to the basement membrane by their egg-stalks. The ripe eggs were about $60{\mu}m$ in diameter and they were surrounded by gelatinous membrane. Most male germ cells in mature stage were transformed into the spermatozoa and they formed the sperm bundles. After spawning, undischarged ripe eggs and spermatozoa remained in the ovarian sac and the testicular tubule respectively for some time, then they finally degenerated. Especially the early spent ovarian sacs in May did not contract significantly and then they took part in the secondary maturation within two or three months during the summer season. The monthly changes of the fatness well agreed with the reproductive cycle. The reproductive cycle of F. mutica could be classified into six successive stages : multiplicative, growing, mature, spent, degenerative and recovery stage. It seems that the spawning season is closely rotated to the water temperature, and the spawning occurs from May to October at about $20^{\circ}C$ in water temperature. The peak spawning seasons appeared twice a year between June and July and in September. Acknowledgement The authors wish to express their gratitude to Dr. Kim, In Bae, Dr. Chun, Seh Kyu and Dr. Yoo, Sung Kyoo of National Fisheries University of Busan and Mr. Min, Byoung Seo of National fisheries Research and Development Agency for their critical reading of the manu script.