• Journal of Aquaculture
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• v.9 no.1
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• pp.83-92
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• 1996

This study was conducted to compare the immunochemical properties of female-specific serum proteins (vitellogenin, VTG) and egg yolk proteins in female fusilier, Caesio diagramma. VTG of fusilier was identified and characterized by using immunochemical analysis. Two types of VTG (VTG1 and VTG2) reacted clearly with antiserum against egg proteins, were confirmed in the serum of mature female. The results of sephacryl S-300 showed that the molecular weights of VTG1 and VTG2 were 560,000 and 410,000, respectively. Yolk proteins, E2 and E3, were isolated from egg extracts, and molecular weights of them were estimated 410,000 and 170,000, respectively. The treatment of $17\beta$-estradiol ($E_2$) to males has induced the synthesis of VTG of which immunological characteristics seems to be similar to the yolk proteins. The results suggest that VTG can be synthesized in the liver by the action of $E_2$ stimulation, and incorporated into the oocytes through the blood circulation. The level of serum $E_2$ was moderately high throughout the spawning period of June. The level of serum VTG was also sustained at high in May and June. The concentration changes of serum $E_2$ and VTG were corelated to the ovarian development in female fusilier. The results indicated that $E_2$ may have some important roles for the vitellogenesis in female fusilier. Also) the VTG can be a precursor protein of yolk not only because it could be synthesized in the liver then incorporated into the oocytes but also because an egg yolk protein had the similiar molecular weights and antigenecity with VTG.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.38 no.5
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• pp.309-315
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• 2005

Changes of sex steroid hormones in the plasma of yellowfin goby, Acanthogobius flavimanus were investigated in relation to the gonadosomatic index (GSI), the hepatosomatic index (HSI) and gonadal development. The GSI in females rose rapidly in November and remained high from December to May $(7.26\pm0.89­6.62\pm0.02)$. The Male's GSI also increased gradually from November and was highest in May $(0.16\pm0.08)$. The HSI in both sexes was in reverse correlation with the GSI, and the HSI was low during the spawning season (February-May). In females, the $estradiol-17{\beta}\;(E_2)$ level increased during vitellogenesis (November and December) and reached its maximum $(8.13\pm2.87 ng/mL)$ at the maturing period, in January. $17{\alpha},\;20{\beta}$-dihydroxy-4-pregnen-3-one$(17{\alpha}20{\beta}OHP)$ gradually increased from October $(0.063{\pm}0.02ng/mL)$ to March $(0.16{\pm}0.02ng/mL)$ and increased rapidly in May. The level of testosterone (T) showed a similar tendency of $E_2$. In males, T increased gradually during spermatogenesis from September to December $(0.14{\pm}0.06­0.26{\pm}0.10ng/mL)$ and peaked in January $(0.36{\pm}0.29 ng/mL)$ when the spermatozoa filled the testis. 11-KT also rose markedly in January and then decreased. On the other hand, $17{\alpha}29{\beta}OHP$ in males did not show any clear tendencies.

• The Korean Journal of Malacology
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• v.20 no.1
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• pp.55-63
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• 2004

Gonadosomatic index, condition index and reproductive cycle with the gonadal development of the female Octopus ocellatus were investigated by histological observations and morphometric data, from January to December, 2000. And changes in biochemical components of the ovary and the trunk tissues including the digestive organ associated with gonadal development were studied by biochemical analysis from January to October, 2001. The specimens were collected at the coastal waters of Buan, Jeollabuk-do, Korea, from January 2000 to October 2001. O. ocellatus is a dioecious organism. The gonad of O. ocellatus locates medially in posterior region of the body. Morphology of the ovary shows round and oval in shape, the average diameter and external colour of ripe ovary was 32 mm and semitransparent light brown in colour. As the ovary was getting mature, transparent elongated eggs covered with chorion were present in the ovarian cavity. Monthly changes in the gonadosomatic index (GSI) showed a similar pattern with those of the condition index. The GSI and condition index began to increase in March and reached the maximum in April. And then, their values decreased from May and reached the minimum in September. Reproductive cycle of O. ocellatus can be categorized into five successive stages: early developing stage (September to December), late developing stage (November to March), ripe stage (March to May), partially spawned stage (April to June), and degenerative/resting stage (June to October). Follicle cells attached to an oocyte were involved in vitellogenesis in the cytoplasm of the vitellogeneic oocyte and formation of chorion (secondary egg membrane) of the ovarian eggs. Spawning occurred between April and June. The spawning period was once a year and the peak took place between May and June. This species belongs to semelparity. According to changes in biochemical contents of the ovary and the digestive organ, monthly variations of moisture, total protein, total lipid and glycogen contents (%) in the ovary showed a negative correlationship with those of the trunk tissues including the digestive organ. Accordingly, it is assumed that the ovary only may be received nutrient supply (total lipid content) for gonadal development from the trunk tissues including the digestive organ (r = -0.55, p < 0.05).

• Journal of Aquaculture
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• v.8 no.1
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• pp.1-19
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• 1995

In red seabream, Pagrus major the female specific protein in the vitellogenic female serum was identified by Ouchterlony's immunodiffusion test and immunoelectrophoresis. The female specific serum protein might be vitellogenin based on the results of the immunological analysis for the male and vitellogenic female sera and crude egg extracts. Also, it was identified by the immunodiffusion test that the purified yolk protein from ovarian egg extracts has antigenic identities shared with the female specific serum protein. To study the relationship between the maturational stages of gonad and plasma levels of vitellogenin, these were measured from the late resting period (January) to the vitellogenic preiod (April) by the modified enzymeimmunoassay (EIA) using antiserum against yolk protein. The level of plasma vitellogenin began to increase in February (previtellogenesis stage) and continuously increased with the ovarian growth during the vitellogenesis period (March to April). The plasma vitellogenin levels were significantly different between the females and the males in February. Validation for the modified EIA system. was tested .The absorbance curve of serial dilutions of serum from the vitellogenic female was paralleled to the standard curve of yolk protein; $109\pm5.6\%$ recovery was achieved by the modified EIA. And the intraassay coefficients of variation were less than 10% within the concentration ranging from 31.3 ng/ml to 1,000 ng/ml. These findings suggest that the sex determination in adult red seabreams could be possible by using the modified EIA as early as in February.

• The Korean Journal of Zoology
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• v.31 no.3
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• pp.165-176
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• 1988

Stmcturai changes at the surfaces of oocytes of Pseudopotamilla occelata were examined by electron microscopy. The oocytes, which grow up to the flnai stage in the same coelomic fluid, once released from the ovary at 5 $\mu$ m stage, change in the structure of the vitelilne envelopes. Microvilli were found to change gready in structure, abundance and behaviour dudng oogenesis. Microvilli are short and bifurcated at the previtellogenic stages and grow in size, but the number increases only during previtellogenesis but decreases during vitellogenesis. Glycocaiyx structures begin to form at the tips of microvitli at the early previtellogenic stages and become more abundant as oocytes grow and remain at the final stage of oogenesis. The tips of microvilli are separated from the stems at the late vitellogenic stages to form vesicles simultaneously with retraction of the microvilli. Vitelline envelope consists of outer, intermediate and inner layers at the previtellogenic stages. However, the inner layer becomes thickened and differendated into two sublayers at 80 $\mu$m stage, - while the outer and intermediate layers remain constant in the thickness. These structural changes were presumably the results of functional differentiation of the vitelline envelope throughout oogenesis even in the same milleu.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.32 no.5
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• pp.624-628
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• 1999

Annual plasma levels of vitellogenin and sex steroids were investigated in relation to the gonadal development for understanding the endocrine control of reproduction in spotted flounder, Verasper variegatus. The plasma vitellogenin level was highest, 6.36 mg/ml, in November when vitellogenesis was most active. The level, thereafter, decreased to 3.81 mg/ml in December with the initiation of spawning. On the other hand, estradiol-17 $\beta$ was highest, 2.7 ng/ml, in December, and rapidly decreased in January when spawning occurred. The decreased level of estradiol-17$\beta$, around 0.2 ng/ml, remained unchanged until May. The profiles of plasma testosterone were similar to those of estradiol-17$\beta$ in the fish, The plasma 17 $\alpha$-hydroxyprogesterone level was relatively low throughout the spawning period, but increased slightly with the initiation of ovarian development, In males, the plasma testosterone and 11-ketotestosterone were highest in December when spermiation actively proceeded, but rapidly decreased during the spawning period (January).

• Applied Microscopy
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• v.31 no.1
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• pp.19-35
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• 2001

Outlines for plasma $estradiol-17\beta$, components, electrophoretic patterns, and ultrastructural changes were obtained in female rainbow trout (Oncorhynchus mykiss) during the seasonal reproductive cycles. Plasma $estradiol-17\beta$ under the natural conditions, exhibited distinct seasonal variation, peaking very late in vitellogenic season during September, decreasing gradually the halt of spawning in December, and ultimately falling during the early stages of seasonal ovarian recrudescence in February and March. This change in $estradiol-17\beta$ appeared to stimulate vitellogenin production as evidenced by increases in plasma calcium, phosphorus, glucose, albumin and total protein levels. The electrophoretic patterns of late maturing or spawning oocytes were stained more intensively than those of late perinucleolus oocytes (molecular weights of approximately 70,000 and 200,000). Two protein bands were found in the SDS-PAGE separation, coincident with the $estradiol-17\beta$ hormone peak. Gonadosomatic indices (GSI) significantly increased from October to January, and showed the highest peak in January, coinciding with the numerically abrupt increase of ripe ova in female. A positive correlation (r=0.701, p<0.01) was established between plasma $estradiol-17\beta$ levels and the gonadosomatic index during the prespawning. The highest level of hepatosomatic index (HSI) observed in December. During the breeding season (December), the gonadotropes were large and filled with GTH-containing inclusions such as granules and globules. The vitellogenic phase began as late perinurleolus oocytes became transformed into early maturing oocytes through the accumulation of yolk, and oocytes reached the late maturing stages as the ooplasm was completely packed with yolk. Marked ultrastructural changed in the granulosa cells during nuclear migration involve the dilation of the rough endoplasmic reticulum and the appearance of the rod-shaped mitochondria with tubular cristae. Microvilli (finger-like projections), from the zona radiata and from the oocyte grew, and made contact with each other in the pore canals of the zona radials during vitellogenesis, but were withdrawn as the zona radiata became more compact and devoid of pore canals during oocyte maturation. The zona radiata grew to a tripartite structure such as an outer thin homogeneous layer, and two inner thick helicoidal layers (zona radials interna and zona radiata externa). Under the normal conditions, the ovarian follicle influenced the histological development and periodical secretion of the hormones , sufficient for a oogenesis and gonadal steroid production.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.33 no.5
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• pp.431-438
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• 2000

To clarify the annual reproductive cycle in a rockfish, Sebastes schlegeli, monthly changes in gonadosomatic index (GSI), hepatosomatic index (HSI) and histological feature of gonads and plasma levels of sex steroid hormones ($estradiol-l7{\beta},\;17{\alpha},\;20{\beta}-dihydroxy-4-pregnen-3-one,\;testosterone\;and\;11-ketotestosterone$) were investigated. The annual reproductive cycle in females could be divided into 5 periods as follows: 1) recovery period (June to September): serum level of $estradiol-l7{\beta}$ increased gradually; 2) vitellogenesis period (Septemer to february) : vitellogenic oocytes were obsewed, GSI sustained high value, and serum level of $estradiol-l7{\beta}$ increased; 3) gestation period (February-April): developing larva showed in the ovary, and serum levels of $17{\alpha},\;20{\beta}-dihydroxy-4-pregnen-3-one$ and testosterone increased; 4) partrition period (April to May) : larva were delivered, and value of GSI and serum levels of hormones decreased rapidly; 5) resting period (May to June) : value of GSI and serum levels of $estradiol-l7{\beta}$ and testosterone remained low. The annual reproductive cycle in males could be divided into 6 periods; 1) early maturation period (April to June): value of GSI and serum levels of hormones incresed gradually, cyst of spermatogonia incresed in number, and a small number of cyst of spermatocyte was observed; 2) mid-maturation perid (June to September); value of GSI and serum levels of hormones increased, and germ cells in many cysts were undergoing active sperrnatogenesis; 3) late maturation period (September to November) : value of GSI and serum levels of hormones remained high and spermatozoa were released into the lumina of the seminal lobules; 3) spermatozoa dischaging period (Nobember to December) : the lumina of the seminal lobules were enlarged and filled with mature spermatozoa; 4) degeneration period (December to Februauy)i value of GSI decresed and cyst of spermatocyte were decresed in number; 5) resting period (December to April) : no histological changes of testes were observed, and value of GSI and serum levels of hormones remained low. In November, the lumina of the seminal lobules were filled with mature spermatozoa and sperm masses were present in the ovarian cavity. Thus, copulation in this species occurred in November and December.