• Journal of Embryo Transfer
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• v.20 no.1
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• pp.1-8
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• 2005

This experiment was carried out to investigate the effects of saccharide in the lactose-egg yolk(LEY) extender for freezing of boar semen on the viability, normal acrosome, fertilizable of in vitro or in vivo oocyte after thawed. Normal acrosome post-thawed spermatozoa was higher when increasing of glucose concentration in LEY extender with 3 or $4\%$ glycerol, but viability was not significant. Viability of the post-thawed spermatozoa was higher when fructose or fructose and glucose were added to LEY extender with $3\%$ glycerol than glucose and sucrose or fructose, glucose and sucrose(P<0.05). Rate of normal acrosome of post thawed spermatozoa was higher when both fructose and glucose$(81.4{\pm}2.3\%)$ were added to the LEY extender than saccharide not added$(41.6\pm0.6\%)$ to it(P<0.001). The percentage of fertilization, cleavage and development to blastocyst of oocytes fertilized with post-thawed spermatozoa from freezing by LEY extender were $70.8\~80.7\%$, $44.6\~45.7$ and $13.6\~16.0\%$, respectively. Conception rate by artificial insemination with frozen boa. semen was higher$(83.1{\pm}0.3\%)$ than commercial frozen semen from SGI company$(50.0{\pm}0.1\%,\;P<0.05)$, but litter size were no significant differences between frozen by LEY extender$(9.4{\pm}1.7\~10.4{\pm}0.7head/sow)$ and SGI semen$(8.0{\pm}1.1 head/sow)$.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.16 no.4
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• pp.389-394
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• 1983

The right eye flounders, Limanda yokohamae Gunther widely distributed in the coastal waters of Korea and Japan. On Feburuary 3, 1983, the authors obtained a number of artificial fertilized eggs from the adult fishes(male: 285mm in total length; female: 297mm) caught by a trawl. The eggs of this species is demersal and adhesive, and the diameter of these eggs was varied in $0.71{\sim}0.80mm$. The egg capsule is colorless and transparent, and the eggs do not contain any oil globules, The hatching took place in 120 hour after fertilization at the water temperature $5.5{\sim}17.0^{\circ}C$. The newly hatched larvae were $2.64{\sim}2.72mm$ in total length with long trunk. Myotome number was $9{\sim}10+30{\sim}32=39{sim}42$ and yellowish brown melanophores were appeared. In 3 days after hatching out. The larvae attained 3.3mm in total length, and the mouth began to move. Xanthophore appeared also on the opereulum at this time. After 7 days the larvae attained 3.70mm in total length, and became the postlarvae absorbing the yolk completely.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.20 no.4
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• pp.308-316
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• 1987

The flying fish, Prognichthys agoo, is widely distributed in the coastal waters of south-eastern Korea. On July 14, 1986, mature adults of flying fish were captured from U-do, Cheju-do. The eggs were stripped and fertilized by the wet method on the ship. The mature eggs are demersal and adhesive with 30-40 filaments. The egg diameter varied from 1.42 to 1.58 mm. The water temperature throughout incubation ranged from 23.70 to $27.82^{\circ}C$, and salinity was maintained at $30.75-33.76\%_{\circ}$. The hatching took place in 174 hours after fertilization. The newly hatched larvae measured 4.75-5.25 mm in total length possessing yolk sac and about 45-46 myotomes. The larvae cultured for ten days after hatching reached 11.45-12.60 mm in total length and entered the juvenile period of life. Twenty days after hatching, the juveniles measured 20.01 mm in mean total length, and the scales were formed behind the pectoral fin.

• Korean Journal of Animal Reproduction
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• v.20 no.2
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• pp.111-117
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• 1996

This experiment was designed to evaluate the effect of transforming growth factor-$\beta$ (TGF-$\beta$) and insulin-like growth factor-I (IGF-I) in bovine oocyte maturation in the presence or absence of serum on subsequent fertilization and embryo development. In addition, various concent rations of these growth factors were evaluated for the ability to promote development of eight-cell stage embryos to the blastocyst stage. Cumulus-oocyte complexes were recovered from 2 to 6 mm follicles obtained from slaughterhouse ovaries and cultured at 38.5$^{\circ}C$ for 24 hours in TCM-199 (HEPES Modification) with or with out 20 % fetal bovine serum (FBS) to which the following growth factors were added TGF$\beta$ IGF-l or TGF $\beta$ + IGF-I, all at 10 ng/ml each. The matured oocytes were fertilized in IVF-TL medium with frozen-thawed semen at a concentration of 1 ${\times}$ 10$^6$ cells/ml of fertilization medium following Percoll separation. After 24 hours of sperm-egg incubation, the embryos were transferred to CZB medium without glucose for 48 hours and then cultured in TCM-199 with 20% fetal bovine serum (FBS) for 96 hours. The addition of growth factors to IVM medium in the presence of serum had no effect on cleavage and subsequent embryo devlopment to blastocyst. In the absence of serum, TGF- improved cleavage and development to blastocyst compared to control's(p<0.05) and no synergistic effeet of IGF-I + TGF-$\beta$ was observed. In the second experiment, eight-cell embryos obtained by in vitro maturation (IVM) in TCM-199 + 20% FBS without growth facrors and in vitro fertil-ization (IVF) were cultured in the in vitro cuiture (IVC) medium supplemented with 5, 10 ng/ml TGF-$\beta$ or 5, 10, 50, 100 ng/ml IGF-I. Cleavage rate and development to the blastocyst stage was observed during seven days of incubation. The supplementation of 10 ng/ml TGF-$\beta$ to lVC medium for eight-cell embryos improved development to blastocyst (p<0.05) compared to control. In conclusion, these data indicate that the supplementation of growth factors to IVM medium in the presence of serum does not influence cleavage and subsequent embryo development. However, significantly more oocytes matured in serum-free TCM-199 and eight-cell embryos cultured in lVC medium developed to blastocyst with supplementation of 10ng/ml TGF-$\beta$.

• Korean Journal of Plant Tissue Culture
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• v.27 no.6
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• pp.423-428
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• 2000

In 1997 when cloned sheep Dolly and soon after Polly were born, it had become head-line news because in the former the nucleus that gave rise to the lamb came from cells of six-year-old adult sheep and in the latter case a foreign gene was inserted into the donor nucleus to make the cloned sheep produce human protein, factor IX, in e milk. In the last few years, once the realm of science fiction, cloned mammals especially in livestock have become almost commonplace. What the press accounts often fail to convey, however, is that behind every success lie hundreds of failures. Many of the nuclear-transferred egg cells fail to undergo normal cell divisions. Even when an embryo does successfully implant in the womb, pregnancy often ends in miscarriage. A significant fraction of the animals that are born die shortly after birth and some of those that survived have serious developmental abnormalities. Efficiency remains at less than one % out of some hundred attempts to clone an animal. These facts show that something is fundamentally wrong and enormous hurdles must be overcome before cloning becomes practical. Cloning researchers now tent to put aside their effort to create live animals in order to probe the fundamental questions on cell biology including stem cells, the questions of whether the hereditary material in the nucleus of each cell remains intact throughout development, and how transferred nucleus is reprogrammed exactly like the zygotic nucleus. Stem cells are defined as those cells which can divide to produce a daughter cell like themselves (self-renewal) as well as a daughter cell that will give rise to specific differentiated cells (cell-differentiation). Multicellular organisms are formed from a single totipotent stem cell commonly called fertilized egg or zygote. As this cell and its progeny undergo cell divisions the potency of the stem cells in each tissue and organ become gradually restricted in the order of totipotent, pluripotent, and multipotent. The differentiation potential of multipotent stem cells in each tissue has been thought to be limited to cell lineages present in the organ from which they were derived. Recent studies, however, revealed that multipotent stem cells derived from adult tissues have much wider differentiation potential than was previously thought. These cells can differentiate into developmentally unrelated cell types, such as nerve stem cell into blood cells or muscle stem cell into brain cells. Neural stem cells isolated from the adult forebrain were recently shown to be capable of repopulating the hematopoietic system and produce blood cells in irradiated condition. In plants although the term$\boxDr$ stem cell$\boxUl$is not used, some cells in the second layer of tunica at the apical meristem of shoot, some nucellar cells surrounding the embryo sac, and initial cells of adventive buds are considered to be equivalent to the totipotent stem cells of mammals. The telomere ends of linear eukaryotic chromosomes cannot be replicated because the RNA primer at the end of a completed lagging strand cannot be replaced with DNA, causing 5' end gap. A chromosome would be shortened by the length of RNA primer with every cycle of DNA replication and cell division. Essential genes located near the ends of chromosomes would inevitably be deleted by end-shortening, thereby killing the descendants of the original cells. Telomeric DNA has an unusual sequence consisting of up to 1,000 or more tandem repeat of a simple sequence. For example, chromosome of mammal including human has the repeating telomeric sequence of TTAGGG and that of higher plant is TTTAGGG. This non-genic tandem repeat prevents the death of cell despite the continued shortening of chromosome length. In contrast with the somatic cells germ line cells have the mechanism to fill-up the 5' end gap of telomere, thus maintaining the original length of chromosome. Cem line cells exhibit active enzyme telomerase which functions to maintain the stable length of telomere. Some of the cloned animals are reported prematurely getting old. It has to be ascertained whether the multipotent stem cells in the tissues of adult mammals have the original telomeres or shortened telomeres.

• Korean Journal of Ichthyology
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• v.12 no.2
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• pp.137-145
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• 2000

The embryonic and larval development of Chelon lauvergnii (Eydoux & Souleyet) was surveyed by incubating artificially inseminated eggs with parent fishes obtained at Kang-wha island in the mid-western coastal area of Korea on June, 1997. The fertilized eggs were transparent, spherical in shape, measuring 0.95~1.08 mm in diameter, having a large oil globule, and their perivitelline space narrow, and began to hatch at 40 hrs. in water temperature $22{\pm}1^{\circ}C$. The newly hatched larvae were 2.35~2.68 mm in total length with 23 myomeres, anus opened, mouth closed, preanal length 58.7~61.6% of total length, oil globule located in posterior end of yolk sac. Melanophores, branch in shape, were distributed mainly along the ventro-lateral region of trunk part and a few on the anterior end of caudal part and surface of oil globule. The larvae measuring 3.08~3.36 mm in total length absorbed yolk material completely in 3 days after hatching, in which air bladder began to appear and mouth opened. In 8 days after hatching, the larva was measured 5.09 mm in total length, its posterior end of notochord began to flex upward and the caudal fin rays differentiated as 7, finfold of the second dorsal and anal fins appeared. In this time, melanophores, branch in shape, were concentrated in the anterior half region of the caudal part and a few also distributed on the top of head, snout region, ventral margin of lower jaw and isthmus region. In 12 days after hatching, the larva measuring 8.48 mm in total length completed all the fins (D. IV-9; P1. 16; P2. I, 5; A. II, 9) and reached to the juvenile stage. Melanophores, in this time, were distributed on the mid-lateral region of the caudal part in enlargment than before and a few also found in the dorso-lateral region of the trunk part, and in the cheek region.

• Korean Journal of Ecology and Environment
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• v.44 no.1
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• pp.1-8
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• 2011

We investigated developmental stages of embryo and early life history of the Korean indigenous fish, the northern loach, Cobitis pacifica in 2009 in order to understand fundamental knowledges for conservation of this species. Eggs were obtained after hormones injections (LHRH-a, HCG) and were artificially fertilized by the dry method. The embryo was spherical, separative demersal, faint white, and averaged $1.09{\pm}0.04\;mm$ (n=20) in diameter. The hatching of the embryo took place in about 48 hours after fertilization under water temperature of $21.0{\sim}24.0^{\circ}C$ and the newly hatched larvae averaged $2.87{\pm}0.05\;mm$ (n=20) in total length (TL). Four days after hatching, the larvae grew up to $6.86{\pm}0.10\;mm$ (n=10) in TL and york sac absorption, mouth and anus opening were shown. Fourteen days after hatching, most of fin-rays appeared at $10.71{\pm}0.34\;mm$ (n=10) in TL and color spots on the body surface were attained. Twenty six days after hatching, the larvae grew up to $14.88{\pm}0.45\;mm$ (n=10) in TL, and all their fin-rays were formed. Therefore, according to current study regarding the morphological development of Cobitis pacidica, the conversion from larval to juvenile stages occurred at 26 days after hatching. Eighty days after hatching, the larvae were $33.3{\pm}1.25\;mm$ (n=10), and their body shape and color pattern were similar to adult fish. In this study, embryonic development and early life history of the northern loach, Cobitis pacifica show morphological characteristics of Cobitidae family. We expected that our results can be used as an fundamental knowledges for restoration study of indigenous fish species.

• The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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• v.6 no.4
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• pp.265-273
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• 2001

The sea urchin has been used as sea food in many countries. This species has also been an important organism of embryological studies for more than a century. In recent years, sea urchin embryos are being used as testing materials for toxicity of pollutants and toxins. Usefulness of sea urchin embryos as experimental models comes from the easiness in obtaining sea urchin samples and a lot of gametes, in rearing embryos in the laboratory, in observing the cellular movement and organ formation during the embryogenesis and in manipulating blastomeres and genetic maferials. The sea urchin in itself is a key organism for the understanding of deuterostome evolution from the protostomes and of indirect development of marine invertebrates which undergo the planktotrophic larval stage. A fertilized sea urchin egg goes through rapid cleavage and becomes a 60 cell embryo 7hr after fertilization. It then develops into a morula, a blastula, a gastrula and finally a pluteus larva approximately 70 hr after fertilization. At the 60 cell stage, the embryo comprises of five territories that express territory-speciflc genes and later form different organs. Micromeres at the vegetal pole ingress into the blastoceol and become the primary mesenchyme cells(PMCs). PMCs express genes involved in skeletogenesis such as SM30, SM37, SM50, PM27, msp130. Among the genes, SM37 and SM50 are considered to be members of a gene family which is characterized by early blastula expression, Glycine-Proline-Glutamine rich repeat structures and spicule matrix forming basic proteins. Genetic studies on the sea urchin embryos help understand the molecular basis of indirect development of marine invertebrates and also of the biomineralization common to the animal kingdom.

• Korean Journal of Animal Reproduction
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• v.24 no.1
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• pp.69-76
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• 2000

This study was carried out to improve of effective culture system on development of IVM/IVF/IVC bovine embryos. The cumulus-oocyte-complexes (COCs) collected from Korean cattle ovaries harvested at a local abattoir were matured in 50 ${mu}ell$ of TCM199 supplemented with 10% fetal bovine serum (FBS) and hormones (35 $\mu\textrm{g}$/$m\ell$ FSH, 10 $\mu\textrm{g}$/$m\ell$ LH, 1$\mu\textrm{g}$/$m\ell$ estradiol 17 $\beta$ under paraffin oil at 39$^{\circ}C$ in a humidified atmosphere of 5% $CO_2$in air. At 24 hrs after culture, matured oocytes were fertilized in vitro for 22~24 hrs with motile semen in which obtained by centrifugation of a frozen thawed semen on Percoll-density gradients (45% vs. 90%) at 500 g for 20 min. The presumptive zygotes were divided into three experimental groups. Single egg (Group 1), 25 (Group 2) or 50 eggs (Group 3) were cultured on cumulus cell in 50 ${mu}ell$ TCM199 supplement with 10% FBS for 6~9 days after fertilization. In vitro developmental rates into the blastocysts in the groups 2 and 3 were significantly (P＜0.05) higher than those of group 1 (37,27 vs. 6%, respectively). Cell number of blastocysts obtained in groups 2 and 3 at day 8 were significantly (P${mu}ell$) resulted in higher developmental competence and cell number of bovine blastocysts produced in vitro than those the culture of single embryos with cumulus cells.

• Korean Journal of Ichthyology
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• v.25 no.2
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• pp.82-89
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• 2013

The spawning behavior, development of eggs and larvae of the Five striped damselfish, Abudefduf vaigiensis were studied. The Five striped damselfish were caught at Dolsan Island, Yeosusi, Jeollanamdo from May in 2011. As a result of observation, male fish attracted female after cleaning the rock. Female left after spawning and male protected their eggs until they had hatched out. The fertilized eggs were elliptical in shape (mean long diameter: 1.06 mm; mean short diameter: 0.55 mm) and transparent. Larvae hatched at 53 hrs after fertilization in $24.5{\sim}26.5^{\circ}C$(mean $25.0^{\circ}C$). The newly hatched larvae were 2.55~2.86 mm (mean 2.71 mm, n=10) in total length and their mouth and anus were already opened. They began to eat rotifer and transformed to postlarva stage. 3 days after hatching postlarva was measured 2.74~2.97mm(mean 2.84 mm, n=10) in total length. 10 days after hatching postlarva was measured 3.85~4.20mm(mean 4.00 mm, n=10) in total length with dosal fin rays IV-5; ventral fin rays I-3; caudal fin rays 1~2.