• Journal of Embryo Transfer
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• v.11 no.1
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• pp.71-80
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• 1996

The envelope of the rnannnalian oocyte plays crucial roles in sperm-oocyte interactions by providing sperm receptors, inducing acrosome reaction and preventing polyspermy. Understanding of properties of the zona pellucida (ZP) is essential for the artificial control of fertility in mammals. This study was carried out to produce and characterize monoclonal antibodies(MAbs) to porcine ZP proteins. Approximately 8,000 ZPs were obtained from follicular oocytes and dissolved in 40$\mu$l of double distilled water. Following immunization through foot-pad injections of Balb /c mice with a ZP solution, the popliteal lymph nodes were recovered at 2 weeks after the last injection. Hybridoma cell lines were established by fusing lymph node cells with P3X63 myeloma cells through selection using HAT medium and screening by immunofluorescence(IF) microscopy on the isolated ZP. Secreted MAbs were found to consist k chains and different heavy chains as evidenced by isotyping. Some of the MAbs demonstrated high specificity to the ZP in IF. The Mabs also showed positive cross reactivity with hamster and mouse eggs, while negative with bovine eggs. The results implicate that the MAbs can be used not only for identification of functional regions of the ZP, but also for elucidation of mechanisms involved in fertilization of mammals. The MAbs will provide basic information on biochemical anatomy of the ZP as well as can be candidates for the future contraceptive vaccines.

• Applied Microscopy
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• v.41 no.3
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• pp.155-161
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• 2011

Three spot gourami (Trichogaster trichopterus Pallas, 1770) is a teleost belonging to Belontiidae. The oogenesis of three spot gourami was investigated by light microscope. The ovary was of light peach color and ellipsoidal shape with the major axis 2 cm and the minor axis 1 cm. Cytoplasm of oogonia was basophilic and many nucleoli were located at inside of nuclear membrane. In primary oocyte, lipid droplets were distributed only in the marginal area first, than at nuclear envelope near. In secondary oocyte, the egg envelope was formed and yolk vesicles was formed in the marginal area. The basophilic substance of cytoplasm was changed to acidic. In case of matured egg, thickness of egg envelope and size of egg were increased. The yolk vesicles were changed to yolk mass in accordance with development. The fertilized eggs were the colorless, transparent, spherical, adhesive and pelagic type. A large oil droplet was located in vitelline membrane of the fertilized egg. In conclusion, the oogenesis of three spot gourami was characterized by the increase in cell size, the formations of lipid droplets and yolk, the decrease of basophilic substance in the cytoplasm, and formation of one large oil droplets.

• Applied Microscopy
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• v.40 no.1
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• pp.9-14
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• 2010

Coreoleuciscus splendidus is a teleost belonging to Gobioninae, Cyprinidae. The oogenesis was investigated by light microscope. The ovary was located between intestine and air bladder, a grayish and ellipsoidal shape with the major axis 20 mm and the minor axis 5 mm. Cytoplasm of oogonia was basophilic and many nucleoli were located at inside of nuclear membrane. In primary oocytes, yolk vesicles were distributed only in the marginal area and egg envelope was not formed on the outside of an egg. In secondary oocytes, the egg envelope was formed and yolk vesicles in the cytoplasm were increased than the earlier stage. The basophilic substance of cytoplasm was changed to acidic. In case of matured egg, thickness of egg envelope and size of egg were increased. The yolk vesicles were changed to yolk mass in accordance with development. In conclusion, the oogenesis of C. splendidus was characterized by the increase in cell size, the formation and accumulation of yolk, and the decrease of basophilic substance in the cytoplasm. The oogenesis of C. splendidus is similar with other Cyprinidae fishes. But further study on ultrastructural study of fertilized egg envelope will be necessary to get the species specificity.

• Applied Microscopy
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• v.41 no.3
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• pp.163-168
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• 2011

Tiger barb (Puntius tetrazona Bleeker, 1855) is a teleost belonging to Cyprinidae. The oogenesis of tiger barb was investigated by light microscope. The ovary was of white and ellipsoidal shape with the major axis 2 cm and the minor axis 1 cm. Cytoplasm of oogonia was basophilic and many nucleoli were located at inside of nuclear membrane. In early stage of primary oocyte, yolk vesicles were distributed only in the marginal area. In secondary oocyte, the egg envelope was formed and yolk vesicles was formed in the cytoplasm. The basophilic substance of cytoplasm was changed to acidic. In case of matured egg, thickness of egg envelope and size of egg were increased. The yolk vesicles were changed to yolk mass in accordance with development. Also, there is not the formation of oil droplets in cytoplasm. In conclusion, the oogenesis of tiger barb was characterized by the increase in cell size, the formation of yolk, the decrease of basophilic substance in the cytoplasm, and formation of yolk mass.

• Development and Reproduction
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• v.12 no.2
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• pp.195-202
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• 2008

The gonadosomatic index, germ cell differentiation, and the ovarian cycle in female jicon scallop, Chlamys farreri were studied by histologic and cytologic observations. In the early vitellogenic oocyte, the Golgi complex, mitochondria and rough endoplasmic reticulum were involved in the formation of lipid droplets. In the late vitellogenic oocyte, exogenous substances, namely, glycogen particles and lipid granular substances appeared in the germinal epithelium passed into the ooplasm through the microvilli of the envelope. Yolk granules and multivesicular bodies were involved in the formation of proteinecious yolk granules in the late vitellogenic oocyte. Vitellogenesis occurrs by endogenous autosynthesis and exogenous heterosynthesis. The auxiliary cells function as nutritive cells in the formation and development of the previtellogenic and early vitellogenic oocytes in their earlr stages. Monthly changes in the gonadosomatic index were closely associated with ovarian developmental phases. The reproductive cycle of this species can be classified into five stages: early active stage (January to March), late active stage (March to April), ripe stage (April to August), partially spawned stage (June to August), and spent/inactive stage (August to December). Spawning occurred from June to August, and the major spawning season was from July to August when the sea water was at high temperature.

• Development and Reproduction
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• v.9 no.2
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• pp.123-134
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• 2005

Germ cell development during oogenesis, ovarian maturation and first sexual maturity in female Ruditapes philippinarum were investigated by cytological and histological observations. R. philippinarum is dioecious. During vitellogenesis, the Golgi complex, glycogen particles, and mitochondria were involved in the formation of lipid droplets and lipid granules in the cytoplasm of the early vitellogenic oocyte. In the late vitellogenic oocyte, cortical granules, the endoplasmic reticulum, and mitochondria were involved in the formation of proteid yolk granules in the cytoplasm. At this time, exogenous lipid granular substance and glycogen particles in the germinal epithelium passed into the oocyte through the microvilli of the vitelline envelope. The spawning period was once a year between early June and early October, and the main spawning occurred between July and August when seawater temperature was approximately $20^{\circ}C$. The reproductive cycle of this species can be categorized into five successive stages: early active stage(January to March), late active stage(Februaryto May), ripe stage(April to August), partially spawned stage(May to October), and spent/inactive stage (August to February). Percentages of female clams at first sexual maturity of $15.1{\sim}20.0mm$ in shell length were 52.6%(50% of the rate of group maturity was 17.83mm in length), and 100% for the clams >25.1mm.

• The Korean Journal of Malacology
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• v.25 no.2
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• pp.145-151
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• 2009

The ultrastructural changes in germ cells during oogenesis of the melania snail, Semisulcospira libertina libertina have been investigated by light and electron microscopy. The ovary is located on the surface of the hepatopancreas in the spiral posterior region. The ovary exhibited greenish color in the gonadal mature season. The ovary was composed of a number of oogenic follicles. Oogenesis was divided into five stages with histological features: (1) oogonia, (2) previtellogenic, (3) initial vitellogenic, (4) active vitellogenic, and (5) mature stages. Oogonia were oval in shape, $4-6\;{\mu}m$ in diameter, and had a large nucleus. Previtellogenic oocytes were about $20\;{\mu}m$ in diameter and the cytoplasm reacted with hematoxylin in H-E satin. Initial vitellogenic stage, oocytes were $60-80\;{\mu}m$ in diameter, and small yolk granules of low electron density are scattered in the cytoplasm. Oocytes in the initial vitellogenic stage were connected with ovarian follicle by egg stalk. Active vitellogenic oocyte were $100-120\;{\mu}m$ in diameter. Electron density, size and quantity of yolk granules that are distributed in the cytoplasm have increased from the previous stage. Result of TEM observations, the oocyte contains well-developed Golgi complex, endoplasmic reticula and tubular mitochondria in the cytoplasm. Cytoplasm of mature oocyte was filled with proteinaceous yolk globules of high electron density. In this stage, the length of microvilli in the egg envelope was approximately $1.1\;{\mu}m$.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2005.07a
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• pp.52-52
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• 2005

Oogenesis, the gonadosomatic index (GSI), reproductive cycle and first sexual maturation of the female Neptunea (Barbitonia) arthritica cumingii have been investigated by light and electron microscope observations. In the early vitellogenic oocyte, the Golgi complex and mitochondria were involved in the formation of glycogen, lipid droplets and yolk granules. In late vitellogenic oocytes, the rough endoplasmic reticulum and multivesicular bodies were involved in the formation of proteid yolk granules in the cytoplasm. In particular, compared with the results of other gastropods, it is a different result that appearances of cortical granules at the cortical layer and microvilli on the vitelline envelope, which is associated with heterosynthetic vitellogenesis, were not observed in vitellogenic oocytes during oogenesis. A mature yolk granule was composed of three components: main body (central core), superficial layer, and the limiting menbrane, Monthly changes in the gonadosomatic index in females were studied in 2002 and 2003 were closely associated with ovarian developmental phases. Spawning occurred between May and August in 2002 and 2003 and the main spawning occurred between June and July when the seawater temperature rose to approximately 18${\sim}$23${\circ}$C. The female reproductive cycle can be classified into five successive stages: early activestage (Septmber to October), late active stage ( November to February), ripe stage (February to June), partially spawned stage (May to Aygust), and recovery stage (June to August).

• Clinical and Experimental Reproductive Medicine
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• v.27 no.3
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• pp.301-305
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• 2000

Globozoospermia is a rare type of teratozoospermia. It occurs in 0.1% of all andrological patient's and used to be considered sterile. Globozoospermic patient has 100% round headed spermatozoa, but the spermiogram is normal. The spermatozoa show oval-shape head, the lack of a nuclear envelope, acrosome, and post acrosomal sheath. Objective: To report that a couple with infertility secondary to globozoospermia received ICSI treatment. Material and Method: Case report Results: In the first trial, fertilization was failed. In the second trial, 40% of oocytes were fertilized and all of these embryos were cleaved, but pregnancy did not achieved. In the third trial, sperm injected oocytes were exposed to 10 ${\mu}M$ calcium ionophore for 15 min. All of the injected oocytes were fertilized and proceeded to develop. Triplet pregnancy was achieved after the transfer of six embryos in their third cycle. One embryo vanished and the remaining twins (female) were delivered at 33 weeks of gestation by Caesarean section. Conclusion: This result shows that assisted activation following ICSI may overcome infertility associated with globozoospermia.

• The Korean Journal of Malacology
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• v.22 no.1 s.35
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• pp.1-11
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• 2006

The gonadosomatic index (GSI), oogenesis and reproductive cycle in female Neptunea (Barbitonia) afhritica cumingii were investigated by light and electron microscope observations. In the early vitellogenic oocyte, the Golgi complex and mitochondria were involved in the formation of glycogen, lipid droplets and yolk granules. Late in the vitellogenic oocytes, the rough endoplasmic reticulum and multivesicular bodies were involved in the formation of proteid yolk granules in cytoplasm. In particular, compared with the results of other gastropods, it showed a different result that appearances of cortical granules at the cortical layer and microvilli on the vitelline envelope, which is associated with heterosynthetic vitellogenesis, were not observed in vitellogenic oocytes during oogenesis. A mature yolk granule was composed of three components: main body (central core), superficial layer, and the limiting membrane. Monthly changes in the gonadosomatic index in females studied in 2004 and 2005 were closely associated with ovarian developmental phases. Spawning occurred between May and August in 2004 and 2005 and the main spawning occurred between June and July when the seawater temperature rose to approximately $18-23^{\circ}C$. The female reproductive cycle can be classified into five successive stages: 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).