• Korean Journal of Fisheries and Aquatic Sciences
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• v.32 no.1
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• pp.62-67
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• 1999

Infection rates of oyster ovarian parasite, Marteiliodes chungmuensis and productivity of the oyster shellstock infected with the parasite were investigated at the main seed collection areas in the southern coast of Korea where the extreme bad seed collection of oyster occurred in 1992 and 1993 to evaluate the cause of the bad seed collection. Additionally, the bacterial flora of the sea water and oyster lana were examined to identify the shellfish larva pathogenic bacteria like Vibrio sp. and Pseueomonas sp. In August 1992 to September 1993, infection rate of oyster ovarian parasite, M. chungmuensis at Tongyong, Kyongsangnam province, and Yosu, Chollanam province where the bad seed collection occurred, were $11.8\~100\%$ and $14.3\~100\%$, respectively. But the parasite was not detected in the shellstock collected at Daechon, Chungchongnam province. While a virus-like particle was identified in the cytoplasm of the egg infected by the parasite. The parasite infected egg was not able to fertilize completely. Uninfected egg in the gonad contaminated by the parasite could be able to fertilize but showed an abnormal development till D-shaped larva and then, died of necrosis after D-shaped lana. And some lana developed from low lipid content egg could not develop to the spat and died after the early umbo stage. The predominant bacteria in the oyster lana collected at bad seed collection areas were Pseudomonas sp. and Pseudomonas like bacteria and the occupancy rates were $53.3\~87.1\%$.

• Development and Reproduction
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• v.6 no.2
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• pp.131-139
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• 2002

Since GnRH and its receptor genes are expressed in the ovary, it has been suggested that ovarian GnRH might be involved in the regulation of ovarian function and the apoptosis of ovarian cells. However, it was not known well on the expression and function of GnRH and its receptor in the corpus luteum. The present study was undertaken to investigate whether GnRH and its receptor are expressed in luteal cells and GnRH has any effect on the apoptosis of luteal cells. Luteal cells obtained from the pregnant rats were cultured and stained for GnRH and its receptor proteins. Cultured luteal cells showed distinct immunoreactivity against both anti-GnRH and anti-GnRH receptor antibodies. In addition, the presence of GnRH receptor protein in cultured cells was confirmed by Western blot analysis. To investigate the effect of GnRH on the apoptosis of luteal cells, luteal cells were cultured in the presence of 10$^{-6}$ M GnRH-agonist(GnRH-Ag) for 3, 8, and 12h. TUNEL assay showed that the number of cells undergoing apoptosis increased 12h after culture(P<0.05). DNA fragmentation analysis confirmed the results such that the cells treated for 12h showed the greatest increase of fragmentation(p<0.05). Further, Western blot analysis of cytochrome c in the mitochondrial and cytoplasmic fractions of the luteal cells showed that GnRH-Ag treatment increased the content of cytochrome c in cytoplasm. These results demonstrate that the luteal cells express GnRH and its receptor and GnRH-Ag treatment induces apoptosis of the luteal cells via mitochondrial release of cytochrome c. The present study suggest that the releasing of cytochrome c from mitochondria might be involved in the luteal cell apoptosis induced by GnRH-Ag.

• Journal of Embryo Transfer
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• v.22 no.1
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• pp.69-74
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• 2007

This study was performed to determine the estrous cycles by macroscopic observation of the ovarian changes using the laparoscopy and to make use of these results for embryo transfer in Korean black goat (Copra hircus aegagrus). Laparoscopic examinations of the ovaries were performed from 2 days after $CIDR^(R)$ removal to 22 days after ovulation. The serial morphological changes of follicles and corpus luteum (CL) were observed. CL was classified corpus hemorrhagicum(CH), corpus luteum (CL) and corpus albicans (CA) by its maturation and regression. On the day before ovulation (Day 0), Graafian follicles (GF) were found on one or both ovaries. On the day (Day 1) and $2^{nd}$day (Day 2) of ovulation, and ovulation depression (OD) and an early stage corpus hemorrhagicum $(CH_1)$ were observed at the site of GF, respectively. On Days 3 to 4, more developed and enlarged corpus hemorrhagicum $(CH_2\;and\;CH_3)$ arised from the ovulation of the GF with well vascularization. On Days 5 to 6, it was identified that mature corpus luteum $(CL_3)$ was grown on the ovary, and fully developed CL with adjacent follicles were occupied most part of the ovary on Days 17 and 18. Then the size of CL was diminished, and completely luteal regression $(CL_1\;or\;CA)$ with new large follicle was identified on Days 20 and 22. From these results, the 4 stages of the estrous cycle in Korean black goats were 1) estrus (Day 0) for 1 day, 2) metestrus $(Day\;1{\sim}4)$ for 4 days (stage of CH development), 3) diestrus $(Day\;5{\sim}16/17)$ for 12 or 13 days (luteal stage), and 4) proestrus $(Day\;17/18{\sim}20/22)$ for 4 or 5 days (stage of luteal regression and follicular growing). Laparoscopy for observation of ovarian changes was invasive than laparotomy. Additionally, it had advantages of reduced adhesion and quick operation time. It was considered that laparoscopic examination of ovarian changes will be useful for embryo transfer in the Korean black goats.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.25 no.5
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• pp.413-431
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• 1992

Gonadal development, fertilization and egg development in the maternal body and reproductive cycle of ovoviviparous rockfish, Sebastes inermis, were investigated histologically. Gonadosomatic index(GSI) of male and female were increased from September and reached maximum values in December. In the male, GSI decreased from January, but in the female maintained high values till February and decreased from March. Hepatosomatic index(HSI) was related to GSI conversely. In both sex, HSI increased from February and reached maximum in August as the gonad were degenerating and resting, and began to decrease from September as gonad were glowing. This ovoviviparous rockfish copulates in December. Fertilization with sperms maintained between ovulated oocytes in the ovary occurs in January mainly. Egg development in the ovarian cavity and discharging of hatched preiarva occurs from January to February. The reproductive cycle includes the successive stages: Growing(September), Mature (October-November), Ripe and Fertilization(Decembr-Janua), Egg development and Discharging of hatched larva(January-February), Degeneration and Resting(February-August). According to the frequency distribution of egg diameter and histological observation, the ovoviviparous rockfish discharged the prelarva at a time in a spawning season. The sexual maturation is first attained at 2 ages. All females and males reaches first maturity at body length of 17.1cm and 15.1cm respectively. The mean number of the embryos increased with the increase of the total length of female.

• Korean journal of applied entomology
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• v.29 no.4
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• pp.217-221
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• 1990

Some biological phenomena of Anagrus incarnatus Haliday, an egg parasitoid of planthopper, were studied under three different constant temperatures. Duration from egg to adult emergence of the parasitoid from the BPH eggs were 21.5, 13.6 and 10.6 days under $20^{\circ}C$, $25^{\circ}C$ and $30^{\circ}C$, respectively. Rate of the development was highly correlated with temperature. The critical temperature was estimated as $10.0^{\circ}C$ and the effective degree days was 210.3 day-degree. Durations for the development of A. incarnatus under 25$\pm$$1^{\circ}C$ (16L : 8D) were 12.4, 12.5 and 12.1 days from eggs of N. lugens, S. furcifera and L. striatellus, respectively. Duration of the development of A. incarnatus from 1, 3, 5, 7 days old BPH eggs were 12.5, 12.1, 12.9 days, respectively. The average longevity of adult was 5.3 days under 25$\pm$$1^{\circ}C$. Number of the ovarian and practically oviposited eggs were 34.8$\pm$28 and 28.3$\pm$0., respectively. Female A. incarnatus laid most of the eggs within few days after the emergence ; over 60% within 24 hours, nearly 90% upto the 2nd day, and nearly 100% upto the 3rd day.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.32 no.2
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• pp.198-203
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• 1999

Gonadal development and reproductive cycle off Gomphina melanaegis collected in the coastal waters of Chumunjin, Korea were investigated monthly from April 1996 to April 1997. G. melanaegis was dioecious, The gonads were located between the digestive diverticula and muscle tissues of the foot, The ovary was composed of a number of ovarian sacs, and the testis was composed of several testicular tubules. The flesh weight rate was reached the maximum in August ($23.0\%$), and then decreased to $19.8\%$ in September. In March, the value was reached the minimum ($17.8\%$) and then increased, The size of mature oocyte was ranged $50\~60\mu$m in diameter and had a germinal vesicle with a nucleolus. Mature oocyte contained a large number of yolk granules and lipid granules in its cytoplasm. The spermatozoon was consisted of a conical nucleus with acrosome, a middle piece containing four mitochondria and proximal and distal centrioles, and a flagellum, Sex ratio (male/female) and minimum size for sexual maturation of G. melanaegis were 0.79 and about 25 mm in shell length, respectively. The reproductive cycle could be classified into five succesive stages: multiplicative (December to March), growing (April and May), mature(June), sprawning (July and August), and degenerative and resting (September to November) stages.

• Journal of Embryo Transfer
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• v.12 no.3
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• pp.293-300
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• 1997

Supplementation of maturation medium with additional granulosa cells has beneficial effect on in vitro maturation of bovine follicular oocytes and their subsequent cleavage and development in vitro. However, maturation system using granulosa cells have some disadvantages that collection of granulosa cells is cumbersome and metabolic activity of the cells is variable according to ovarian cycle or follicular size. We hypothesized that bovine immsture oocytes matured without granulosa cell coculture can fertilize and develop normally if the medium volume per oocyte is reduced during in vitro maturation. Immature oocytes were matured for 24 hours in a TCM199 containing 10% fetal calf serum, anterior pitultary hormone (0.02 AU /ml Antrinⓡ) and estradiol with or without granulosa cells in vitro. In Group 1, 35 to 40 oocytes were matured in a well of 4-well plastic dish containing 500 $\mu$l of maturation medium and granulosa cells, and 9 to 10 oocytes were matured in a 50-$\mu$l drop of maturation medium without granulosa cells in Group 2. After maturation, oocytes were coincubated with sperm for 30 hours in a modified Tyrode's medium (IVF). Inseminated oocytes were cultured in a microdrop (30 $\mu$l) of a synthetic oviduct fluld medium (SOFM) containing BSA, Minimum Essential Medium essential and non-essential amino acids for 9 days. As a preliminary experiment, we investigated the beneficial effect of granulosa cells during maturation on subsequent cleavage and development using the same type of culturedishes (4-well dish). Granulosa cells could not increase embryo cleavage after fertilization but significantly improved (p<0.05) embryo development to expanding blastocyst (Table1 and 2). In Group 1, 68 and 80% of inseminated oocytes have cleaved at 30 hours and 2 days after IVF, respectively, which is similar (p>0.05) to the result of Group 2 (69% at 30 hours and 78% at 2 days after IVF). The oocytes in Group 2 showed 21 and 11% of developmental rates to expanding and hatching blastocysts, respectively, which was not significantly different (p>0.05) from those (20 and 10%, respectively) of oocytes in Group 1. In conclusion, it has been clarified that a microdrop culture system without granulosa cells for in vitro maturation can support bovine embryonic development to blastocyst in vitro as readily as a granulosa cell coculture system.

• 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).

• Korean Journal of Fisheries and Aquatic Sciences
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• v.10 no.2
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• pp.71-96
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• 1977

Early development Linuparus trigonus(von Siebold) has been studied based on the samples collected monthly in Je-ju Island, Korea from February, 1975 to January, 1977. Gametogenesis, reproductive cycle, embryonic development were investigated by histological mettled, and morphological description was made on the first phyllosoma larva which reared in the laboratory. Testis is composed of two tubular duct which are symmetrical with H-shaped appearance. Outer layer of testis is of fibrous connective tissue capsule. In the lumen there is a convoluted seminiferous tubule with interstitial tissue. Ovary is a pair of symmetrical blind tubular lobes, and the midportions are connected each other. The ovary consists of a couple of ovarian sacs partitioned by two-layered connective tissue fibers. Proliferation of spermatogonia are observed all the year around on the germinal epithelium of seminiferous tubule. Partial spermatogenesis is always in progress, and the spermatozoa appear all the year around in the tubules. Nutrition of early oogonia is supplied by fibrous mesenchyme which is abundantly distributed in ovarian sacs. Oocytes grow and couplete maturation divisions in the follicle layers. They finally develop into mature ova before spawning. Reproductive cycle is classified into four successive stages; multiplication stage from September to December, growing stage from January to March, maturation division stage from April to May and mature stage from June to August. Spawning takes place from May to August with peak spawning from Into July to early August. Cleavage type is superficial. Blastopore is formed in blasto-disc region which is proliferation of blastoderm cells. Germinal layers are also derived from tile region. Mesoderm formation is originated from endodermal cells which are formed front the blasto-disc region. The endodermal cells are separated by the process of delamination from yolk sac and take part in the formation of the mid-gut. Morphological characteristics of first phyllosoma larva are different from the larvae of other Palinurid and Scyllarid species.

• Journal of Embryo Transfer
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• v.10 no.2
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• pp.105-113
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• 1995

This study was conducted to examine the efficiency of enucleation and blastomere isolation from recipient oocytes and donor embryos, respectively and to determine the effect of oocyte age and electric voltage on the fusion rate and in vitro development of the fused oocytes in rabbit nuclear transplantation. Immature oocytes collected from ovarian follicles were matured in vivo for 12 h in TCM-199 containing FCS and hormones and in vivo matured oocytes were collected 17 to 18 h post-HCG. The fresh and frozen donor embryos of 8- to 16-cell stage were collected from the oviduct of superovulated does. The proportion of successfully enucleated oocytes was greatly lower in in vitro matured oocytes (42.3%) than that (62.7%) in in vivo matured oocytes The level of cytochalasin B for in vivo matured oocytes did not affect the efficiency of enuleation, but 7.5 $\mu$g /mL cytochalasin B for in vitro matured oocytes showed a high enucleation rate significantly. The isolation efficiency of a single blastomere nucleus did not differ between 8- and 16-cell stage embryos. The percentage of single blastomeres isolated from 16-cell stage fresh embryos after 0.5% pronase treatment was greatly higher at 16-min treatment (94.4%) than at 8-min(78. 1%) and the blastomeres(61.5%) isolated from frozen-thawed embryos after 16-min pronase were significantly fewer than those of fresh embryos. The age of recipient oocytes affected nuclear fusion rate. The reconstituted oocytes fused at 24-h age showed slightly higher fusion rate (77.8%) than those (65.0%)fused at 18-h age. The fusion rate of in vitro and in vivo matured oocytes inserted with fresh blastomere did not differ among electric voltages, but the cleavage rate and development to morula-blastocysts of in vitro matured oocytes was more higher under 0.6 kV/cm than under 0.8 to 1.2 kV/cm, while the cleavage rate and development of in vivo matured oocytes was higher under 0.8 to 1.0 kV/cm than under 1.2 kV/cm. The fusion and cleavage rate fol1owing insertion with frozen-thawed blastomere was not different between the in vitro and in vivo matured oocytes and was similar to those from fresh blastomere insertion.