• Korean Journal of Ichthyology
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• v.13 no.1
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• pp.19-23
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• 2001

The full-grown oocyte envelope of the two spined loaches, Niwaella multifasciata and Kichulchoia brevifasciata, was examined by electron microscopy. The oocyte is surrounded by its envelope, and an external modification (an adhesive structure) in the surface of the envelope is present. The envelope consists of two layers, a zona radiata externa which is the site of the adhesive structure, and a zona radiata interna, which has heterogeneous, electron-dense multi layers. The surface (zona radiata externa) of the envelope in N. multifasciata is equipped with short villuslike protuberances, which have a length of 1.5~2.5 ${\mu}m$ and are separated from each other by a distance of 2~2.5 ${\mu}m$. In contrast, K. brevifasciata has undulating or wave-like structures that extend over the entire oocyte surface. The waves are 1.5~2.5 ${\mu}m$ in length and are separated a distance of 2.5~3.3 ${\mu}m$ from each other.

• Applied Microscopy
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• v.29 no.3
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• pp.343-351
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• 1999

The structures of the fertilized egg envelope from three species, three-spot gourami (Trichogaster trichopterus), pearl gourami (Trichogaster leeri) and marble gourami (Trichogaster trichopterus trichopterus) belong to Belontiidae were observed, utilizing light, scanning and transmission electron microscopes. In all three species, 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. The egg envelopes have a single micropyle, which is thought to the pathway of sperm in the area of the animal pole. Specially, the micropyle of three-spot gourami was similar to that of marble gourami which is subspecies of three-spot gourami. An outer surface of the fertilized egg envelope was arranged by grooves in all three species. The fertilized egg envelopes consists of two distinct layers; an adhesive outer layer and an inner layer with high electron density. In conclusion, the morphological similarity of the fertilized egg, micropyle, outer surface and transverse section of the fertilized e9g envelope seems to be an indication of the Belontiidae.

• Korean Journal of Ichthyology
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• v.26 no.3
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• pp.179-184
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• 2014

A histological study on the egg envelope and oogenesis of Gobiobotia brevibarba (Pisces, Cyprinidae) was carried out by a light microscope and a scanning electron microscope. Various developmental cells appeared in the ovary caught during May 2014, spawning season. For the relative area of oocyte, the ovary consisted of mature stage (74.5%), a vitellogenic stage (yolk granule stage, 16.6% and yolk vesicle stage, 6.6%) and previtellogenic stage (perinucleolus stage 2.2%), which means its spawning season. The cytoplasm of the perinucleolus oocyte is acidic and many nucleoli are located at the inner side of the nuclear membrane. The yolk vesicles, an early vitellogenic stage, has a follicular layer and a zona radiata clearly. Numerous villi, called an egg envelope, begin to form on the zona radiata. The yolk granules, an another vitellogenic stage, proceeds and they show a strong eosinophilic nature. Such yolk granules appeared between the yolk vesicles occupying most cytoplasm, and as the stage proceeds, there are some yolk masses fused with each other. Egg envelope is covered with plenty of villi ($2{\sim}3{\mu}m$ in the length) over the entire egg surface.

• 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.38 no.4
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• pp.315-319
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• 2008

Glow-light tetra, Hemigrammus erythrozonus is a teleost belonging to Characidae. The natural habitate of this fish is the wild in the Essequibo river, Guyana and South America. The oogenesis of glow-light tetra was investigated by light microscopy to compare with those of other families. A pair of ovary was located between swim bladder and intestines. The ovary was of white color and ellipsoidal shape with the major axis 11 mm and the minor axis 4 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 amount of basophilic substance was decreased. In case of matured egg, thickness of egg envelope and size of egg were increased, basophilic substance was distributed in only around the egg envelope. The yolk vesicles were changed to yolk mass in accordance with development. In conclusion, the oogenesis of glow-light tetra, Hemigrammus erythrozonus was characterized by the increase in cell size, the formation and accumulation of yolk, and the decrease of basophilia a in the cytoplasm. The oogenesis of glow-light tetra seems to share common patterns in Characidae, teleost and have a similar pattern with other teleost.

• Applied Microscopy
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• v.41 no.3
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• pp.189-196
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• 2011

Histological study on egg envelope and oogenesis of Kichulchoia multifasciata (Pisces, Cobitidae) was carried out by light microscopes and scanning electron microscopes. Various developmental cells appeared in ovary of the specimen catched during November 2010. The cytoplasm of oogonia was acidic and many nucleoli were located at the inner side of nucleus membrane. The size of the oogonia was $103.9{\pm}24.7$ ${\mu}m$ with nucleus size $42.9{\pm}6.9$ (31.1~50.3) ${\mu}m$. Primary oocyte having $277.5{\pm}60.5$ (216.7~354.9) ${\mu}m$ in diameter began to accumulate yolk vesicles. As the developmental stages proceed, secondary oocyte grows larger to $617.6{\pm}85.1$ (503.4~723.6) ${\mu}m$, and eosinophilic yolk granules yolk granules appeared between the yolk vesicles occupying most cytoplasm, and there are some yolk mass formed already. There are some yolk mass formed already. Envelope of fertilized egg investigated by a scanning electron microscope had plenty of microvilli (2~3 ${\mu}m$ in length) over the entire egg surface and a micropyle. Especially, the microvilli surrounding the micropyle were longer than those of egg surface with $5.26{\pm}1.22$ ${\mu}m$.

• 제어로봇시스템학회:학술대회논문집
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• 2003.10a
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• pp.2451-2456
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• 2003

In biological cell manipulation, manual thrust or penetration of an injection pipette into an egg is currently performed by a skilled operator, relying only on visual feedback information. Massive load of various micro injection of either genes, fluid or cells in the postgenomic era calls a more reliable and automatic micro injection system that can test hundreds of genes or cell types at a single experiment. We initiated to study cellular force sensing in zebrafish eggs as the first step for the development of a more controllable micro injection system by any inexperienced operator. Zebrafish eggs at different developmental stages were collected and an integrated biomanipulation system was employed to measure cellular force during penetrating the egg envelope, the chorion. First of all, the biomanipulation system integrated with cellular force sensing instrument is implemented to measure the penetration force of cell membranes and characterize mechanical properties of zebrafish embryo cells. Furthermore, implementation of cellular force sensing system and calibration are presented. Finally, the cellular force sensing of penetrating cell membranes at each developmental stages was experimentally performed. The results demonstrated that the biomanipulation system with force sensing capability can measure cellular force at real-time while the injection operation is undergoing. The magnitude of the measured force was in the range of several hundreds of uN. The precise real-time measurement should provide the first step forwards for the development of an automatic and reliable injection system of various materials into biological cells.

• Korean Journal of Ichthyology
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• v.29 no.4
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• pp.252-257
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• 2017

The oogenesis of the Microphysogobio yaluensis was investigated using light microscopy. Various developmental oocytes appeared in the ovary of the M. yaluensis. The oogenesis is largely divided into four stages: nuclear-chromatin stage, peri-nucleoli stage, vitellogenesis (yolk vesicle and yolk granule stages), and mature stage. The nuclear-chromatin is distributed in a large germinal vesicle as threads. The peri-nucleoli stage has many acidic nucleoli lining at the inner side of the nuclear membrane and an egg envelope just weakly starts. As the oogenesis gradually proceeds, they change to the vitellogenesis stage. The oocyte become to drastically increase and the marginal area of the ooplasm is covered with many vacuoles showing no negative reactions with hematoxylin and eosin staining, called the yolk vesicle stage. Many yolk vesicles-owned oocyte largely increase and as the development continues, its ooplasm is changed from the yolk vesicles to the yolk granules of eosinophilic. At the mature stage, lots of granules merged into a big yolk mass, acidophilic. Even at the mature stage, the egg envelope was still thin between the ooplasm and the follicular layer of the oocyte.

• Applied Microscopy
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• v.29 no.2
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• pp.177-187
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• 1999

Ultrastructure of gametes in the three-spine stickleback, Gasterosteus aculeatus aculeatus was observed, utilizing light, scanning and transmission electron microscopes. The egg of three-spine stickleback is spherical and demersal type. The eggs are highly adhesived to each other but not to substrates. There are many oil droplets in vitelline membrane. The outer surface of egg envelope is arranged by mushroom-like structures and pore canals. The egg have a micropyle, sperm entry site, in the area of the animal pole. The egg envelope consists of three layers, an outer layer with high electron density, a middle layer consisting two layers and an inner layer consisting of 16 to 20 layers. In the fertilized egg envelope, the molecular weights of these components ranged from 14 kDa to 205 kDa. The molecular weights of nam protein bands are 19.4 kDa, 36.7 KDa, 39.4 kDa, 42.9 kDa, 46.1 kDa and 53.0 kDa. The head of spermatozoa is spherical shape and the acrosome is absent. The mitochondria in midpiece are arranged from one to three layers and separated from the axoneme by the cytoplasmic canal. The tail has two lateral fins and the axoneme is of the 9+2 structure.

• The Korean Journal of Malacology
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• v.2 no.1
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• pp.10-12
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• 1986

Pettancylus nipponicus (freshwater limpet) collected from the Lake Choon Cheon were cultrued in aquarium and studied in the respects of an egg deposition and the developmental characters of cleavage. The species were collected only in the Lake Uiam and Choon Cheon and easily sampled on the debris like vinyls and cans. Those smooth surfaces were suitalble substrata for attaching. P. nipponicus, about 4 mm in adult, deposited an egg in one calcareous egg-capsule on the aquarium walls, while many eggs are surrounded in a jelly-like egg-envelope in the most gastropods. And another special features was that an egg-capsule was consisted of two valves and the suture was opended as hatching. A diameter of its egg was $55{{\mu}m}$ and the length of an egg capsule was $620{{\mu}m}$ and $510{{\mu}m}$ in long and short diameter, respectively. It took about 8 hours from beginning cleavage to 16-cell stage and about one year to become an adult.