• Korean Journal of Animal Reproduction
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• v.23 no.2
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• pp.133-139
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• 1999

This study was carried out to investigate in vitro/in vivo development of vitrified-thawed immature mouse oocytes. Immature mouse oocytes were vitrified with EFS40 (40% ethylene glycol, 18% ficoll and 0.5 M sucrose). Thawed oocytes were matured for 16 hr in vitro. Matured oocytes with the first polar body were fertilized with the concentration of 1~2$\times$10$^{6}$ $m\ell$ of epididymal sperm. After fertilization, cleavage ($\geq$ 2-cell) and in vitro/in vivo development rates were examined. $\pi$ Ie results were summarized as follows: in vitro maturation rate of immature mouse oocytes in vitrified-thawed group was similar to that in exposed group (67.5%) and control (66.3%), but cleavage rate of vitrified-thawed oocytes (64.9 %) and blastocyst formation rate (59.0%) were significantly different compared to those of exposed group (83.7 and 74.7%) and control (90.7 and 83.7%) (p＜0.05). However, when the blastocysts derived from immature mouse oocytes vitrified-thawed were transferred to pseudopregnant mouse, total implantation (31.3%) was slightly lower than that in control (40.8%), but live fetus formation rate (66.7%)was slightly higher than that in control (58.1%), there was not significantly different. Therefore, when the blastocyts produced in vitro were transferred into recipients, although the development in vitro of oocytes vitrified-thawed was decreased, live fetus formation rate was similar to that of control group. The present results indicate that immature mouse oocytes can be frozen successfully by vitrification with EFS40.

• Korean Journal of Animal Reproduction
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• v.23 no.3
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• pp.271-278
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• 1999

This study was conducted to investigate the cytogenetic properties, in vitro development, and their relationship in the bovine reconstituted embryos following cell cycle-controlled nuclear transfer. Sixteen-cell stage embryos were treated by nocodazole, and after release from nocodazole treatment, their blastomeres were separated and allowed to subsequent cleavage. Blastomeres within 1.5 h post cleavage(hpc) and at 3hpc were transferred to enucleated oocytes at MII-phase or S-phase. Donor nuclei transferred into M II-phase recipients underwent various nuclear remodeling, such as extrusion of a polar body(PB)-like structure, premature chromosome condensation(PCC) and chromatin modifications. These nuclear remodeling patterns varied by the time post cleavage of donor blastomeres. Developmental rate to the blastocyst stage differed with time post cleavage of donor blastomeres and existence of a PB-like structure. Whereas do-nor nuclei transferred into S-phase oocytes did not undergo PCC and other major modifications, and their developmental potentials less depended on the nuclei types. This result confirms that the nuclear remodeling type differs with donor and recipient cell cycle stage, which affect the development of reconstituted bovine embryos.

• Investigative Magnetic Resonance Imaging
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• v.4 no.1
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• pp.42-51
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• 2000

Purpose: Projection-type Fast Spin Echo (PFSE) imaging is robust to patient motion or flow related artifact compared to conventional Fast Spin Echo (FSE) imaging, however, it has difficulty in controlling $T_2$ contrast. In this paper, Tz contrast in the PFSE method is analyzed and compared with those of the FSE method with various effective echo times by computer simulation. The contrasts in the FSE and PFSE methods are also compared by experiments with volunteers. From the analysis and simulation, it is shown that ${T_2}-weighted$ images can well be obtained by the PFSE method proposed. Materials and methods: Pulse sequence for the PFSE method is implemented at a 1.0 Tesla whole body MRI system and $T_2$ contrasts in the PFSE and FSE methods are analyzed by computer simulation and experiment with volunteers. For the simulation, a mathematical phantom composed of various $T_2$ values is devised and $T_2$ contrast in the reconstructed image by the PFSE is compared to those by the FSE method with various effective echo times. Multi-slice ${T_2}-weighted$ head images of the volunteers obtained by the PFSE method are also shown in comparison with those by the FSE method at a 1.0 Tesla whole body MRI system. Results: From the analysis, $T_2$ contrast by the PFSE method appears similar to those by the FSE method with the effective echo time in a range of SO-lOOms. Using a mathematical phantom, contrast in the PFSE image appears close to that by the FSE method with the effective echo time of 96ms. From experiment with volunteers, multi-slice $T_2-weighted$ images are obtained by the PFSE method having contrast similar to that of the FSE method with the effective echo time of 96ms. Reconstructed images by the PFSE method show less motion related artifact compared to those by the FSE method. Conclusion: The projection-type FSE imaging acquires multiple radial lines with different angles in polar coordinate in k space using multiple spin echoes. The PFSE method is robust to patient motion or flow, however, it has difficulty in controlling $T_2$ contrast compared to the FSE method. In this paper, it is shown that the PFSE method provides good $T_2$ contrast (${T_2}-weighted$ images) similar to the FSE method by both computer simulation and experiments with volunteers.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.32 no.6
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• pp.737-747
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• 1999

To estimate biomass and distribution of the Antarctic krill (Euphausia superba), hydroacoustic survey was conducted on board of R/V Yuzhmorgeologiya, which was chartered by Korea Antarctic Research Program (KARP) group from 18 to 21 December 1998, in the northern part of the South Shetland Islands, Antarctic Ocean, The scientific echo sounder (towing body type) used was EK- 500 (SIMRAD, Norway) with echo integrator (BI-500) at 38 kHz frequency and recorded mean backscattering cross-section coefficient (SA) per 1 $mile^2$ of sea surface. Also, Bongo net sampling was carried out to determine the size of krill and CTD (Conductivity, Temperature and Depth) casting to understand physical structure. Water column was divided into 5 layers (22$\~$65 m, 65$\~$115 m, l15$\~$65 m, 165$\~$215 m and 215$\~$315 m) to know vertical distribution of krill biomass. The standard length of krill collected was between 30 mm and 51 mm, and adult krill had single mode (41 mm). Maximum horizontal length of krill patch was about 35 nautical mile and vertical thickness was about 275 m. High density of krill was appeared in frontal area between Circumpolar Deep Water (>$1^{\circ}C$) and very low temperature water mass (< $-0.5^{\circ}C$) that originate from Weddell Sea. According to the results calculated using target strength equation, krill density was totally higher in continental slope and open water areas than in coastal area. In the study area, krill seems to distribute in depth; density was low at first layer ($\={\rho}=17.0\;g/m^2$) and higher at fourth layer ($\={\rho}=40.19\;g/m^2$). The estimated krill biomass at total survey area and water column was about 2.77 million metric ion ($\={\rho}=151.0\;g/m^2$) and coefficient of valiance ( CV, $\%$) was 19.92. The proportions and biomass of krill biomass at each layer were as follows; layer 1 ($11.3\%$, 0.31 million metric ton, CV=16.24), layer 2 ($13.3\%$, 0.37 million metric ton, CV=34.91), layer 3 ($23.7\%$, 0.66 million metric ton, CV=41.5), layer 4 ($26.6\%$, 0.74 million metric ton, CV=27.84) and layer 5 ($25\%$, 0.69 million metric ton, CV= 26.83).

• Korean Journal of Agricultural Science
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• v.33 no.1
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• pp.35-41
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• 2006

This study was conducted to investigate the developmental ability of interspecies cloned embryos after nuclear transfer of goat fetal fibroblast cells into porcien oocytes. Recipient porcine and goat oocytes were obtained from slaughterhouse and matured in vitro according to established protocols. Enucleation was accomplished by aspirating the first polar body and cytoplasm and a single donor cell was individually microinjected into vitelline space of the enucleated oocyte. The reconstructed oocytes were electrically fused with 0.3M mannitol fusion medium. After electro-fusion, interspecies reconstituted embryos were cultured in PZM-3 for 7 days. In porcine interspecies nuclear transfer with goat fetal fibroblast cells, the cleavage rate of reconstituted embryos were 58.9% which was no significant different from that in porcine nuclear transfer embryos (67.4%). However, the developmental rate into blastocyst stage was 5.4% in interspecies nuclear transfer which was significantly lower than that in porcine intraspecies nuclear transfer (13.6%). When the developmental ability of porcine interspecies nuclear transfer with goat cells was compared with goat intraspecies nuclear transfer, the cleavage rate of embryos were 59.2% and the developmental rate into morular and blastocyst stage was 13.6% in interspecies nuclear transfer which were significantly lower than those in intraspecies nuclear transfer embryos. This result indicated that porcine interspecies nuclear transfer with goat fetal fibroblast cells showed the developmental potential in vitro with lower cleavage and developmental rate compared with intraspecies nuclear transfer.

• Korean Journal of Animal Reproduction
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• v.26 no.3
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• pp.235-243
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• 2002

This study was designed to examine the ability of the bovine (MII) oocytes cytoplasm to support several mitotic cell cycles under the direction of differentiated somatic cell nuclei of bovine, porcine, mouse and human. Bovine GV oocytes were matured in TCM-199 supplemented with 10% FBS. At 20h after IVM, recipient oocytes were stained with 5 $\mu\textrm{g}$/$m\ell$ Hoechst and their 1st polar body (PB) and MII plate were removed by enucleation micropipette under UV filter. Ear skin samples were obtained by biopsy from an adult bovine, porcine, mouse and human and cultured in 10% FBS added DMEM. Individual fibroblast was anlaysed chromosome number to confirm the specificity of species. Nuclear transferred (NT) units were produced by electrofusion of enucleated bovine oocytes with individual fibroblast. The reconstructed embryos were activated in 5 $\mu$M ionomycin for 5 min followed by 1.9 mM 6-dimethylaminopurine (DMAP) in CR1aa for 3 h. And cleaved NT embryos were cultured in CR1aa medium containing 10% FBS on monolayer of bovine cumulus cell for 8 days. Also NT embryo of 4~8 cell stage was analysed chromosome number to confirm the origin of nuclear transferred somatic cell. The rates of fusion between bovine recipient oocytes and bovine, porcine, mouse and human somatic cells were 70.2%, 70.2%, 72.4% and 63.0%, respectively. Also, their cleavage rates were 60.6%, 63.7%, 54.1% and 62.7%, respectively, there were no differences among them. in vitro development rates into morula and blastocyst were 17.5% and 4.3% in NT embryos from bovine and human fibroblasts, respectively. But NT embryos from porcine and mouse fibroblasts were blocked at 16~32-cell stage. The chromosome number in NT embryos from individual fibroblast was the same as chromosome number of individual species. These results show that bovine MII oocytes cytoplasm has the ability to support several mitotic cell cycles directed by newly introduced nuclear DNA.

• Korean Journal of Animal Reproduction
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• v.26 no.3
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• pp.245-252
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• 2002

This study was to evaluate the in vitro survival of bovine enucleated MII (eMII) oocytes according to minimum volume cooling (MVC) freezing method and activation timing, and their in vitro development after somatic cell nuclear transfer (SONT). in vitro matured bovine oocytes for 20 h were stained with 5 $\mu\textrm{g}$/$m\ell$ Hoechst, and their 1st polar body and MII plate were removed by enucleation micropipette under UV filter. Also, eMII oocytes were subjected to activation after (group II) and before (group III) vitrification in 5 ${\mu}{\textrm}{m}$ ionomycin added CRlaa medium for 5 min. For vitrification, eMll oocytes were pretreated with EG10 for 5 min, exposed to EG30 for 30 sec and then directly plunged into L$N_2$. Thawing was taken by 4-step procedures at 37$^{\circ}C$. Survived eMII oocytes were subjected to SONT with cultured adult bovine ear cells. Reconstructed oocytes were cultured in 10 $\mu\textrm{g}$/$m\ell$ of cycloheximide and 2.5 $\mu\textrm{g}$/$m\ell$ of cytochalasin D added CRlaa medium for 1 h, and then in 10 $\mu\textrm{g}$/$m\ell$ of cycloheximide added CRlaa medium for 4 h. Subsequently, the reconstructed oocytes were incubated for 2 days and cleaved embryos were further cultured on cumulus-cell monolayer drop in CRlaa medium for 6 days. Survival rates of bovine vitrified-thawed eMII oocytes in group II (activation after vitrification and thawing) and III (activation before vitrification) were 81.0% and 84.9%, respectively. Fusion rates of cytoplasts and oocytes in group II and III were 69.0% and 70.0%, respectively, and their results were not different with non-frozen NT group (control, 75.2%). Although their cleaved rates (53.4% and 58.4%) were not different, cytoplasmic fragment rate in group II (32.8%) was significantly higher than that in group III (15.6%)(P<0.05). Also, subsequent development rate into >morula in group II (8.6%) was low than that in group III(15.6%). However, in vitro development rate in group III was not different with that in control (24.8%). This result suggested that MVC method was appropriate freezing method for the bovine eMII oocytes and vitrified eMII oocytes after pre-activation could support in vitro embryonic development after SONT as equally well as fresh oocytes.

• Ocean and Polar Research
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• v.28 no.4
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• pp.407-413
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• 2006

The effect of water temperature and photoperiod on the oxygen consumption of the fasted juvenile parrot fish, Oplegnathus fasciatus was investigated to provide empirical data for the early-stage culture management and bioenergetic growth model of the species. The mean body weight of the juvenile used for the experiment was $21.5{\pm}1.9g$, and the oxygen consumption rate was measured under four water temperatures (10, 15, 20 and $25^{\circ}C$) and three photoperiods (24L:0D, 12L:12D and OL:24D) with an interval of 5 minutes for 24 hours using a continuous flow-through respirometer. In each treatment three replicates were set up and 15 juveniles were totally involved. The oxygen consumption rates increased with increasing water temperature under all photoperiod treatments (P<0.001). Mean oxygen consumption rates at 10, 15, 20 and $25^{\circ}C$ ranged $202.1{\sim}403.4,\;306.7{\sim}502.2,\;536.7{\sim}791.0\;and\;879.9{\sim}1,077.4mg\;O_2\;kg^{-1}h^{-1}$, respectively. $Q_{10}$ values ranged $1.58{\sim}2.30$ between 10 and $15^{\circ}C,\;2.44{\sim}3.06$ between 15 and $20^{\circ}C\;and\;1.86{\sim}2.6y9$ between 20 and $25^{\circ}C$, respectively. Mean oxygen consumption rates of O. fasciatus were the highest in continuous light (24L:0D) followed by 12L:12D and 0L:24D (P<0.001). The oxygen consumption of fish exposed to the 12L:12D photoperiod was significantly higher during the light phase than during the dark phase under all temperature treatments (P<0.001). In summary, oxygen consumption rates of the juvenile parrot fish increase with increasing water temperature and lengthening daylight period; and, thereby, changes in water quality resulted from the depletion of oxygen under high temperature and long daylight photoperiod conditions should be monitored.

• Reproductive and Developmental Biology
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• v.29 no.3
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• pp.175-180
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• 2005

This study was conducted to examine the effects of demecolcine-assisted enucleation and recipient cell cycle stage on the development of bovine somatic cell nuclear transfer (NT) embryos. In vitro cultured oocytes for $16\~20$ h were classified by first polar body (1st PB) extrusion and cell cycle stage (MI and MII) and treated $0.4\;{\mu}L/mL$ demecolcine for 40 min before enucleation. Enucleated oocytes were fused electrically with bovine ear skin cells, activated by Ca-ionophore+DMAP, and cultured in vitro. Most of eggs ($86.2\%$) treated with demecolcine protruded a chromosome mass and enucleated efficiently ($98.8\%$, (P<0.05). Demecolcine did not have a deteriorative effect on the development of NT embryos. Developmental rate of NT embryos reconstituted with oocytes extruded 1st PB significantly higher than that of NT embryos produced by oocytes without 1st PB ($18.2\%\;vs.\;4.6\%\cdot$, P<0.05). Cleavage and blastocyst formation rate of embryos reconstituted with MI oocytes ($69.4\%\;and\;5.9\%$, respectively) were significantly lower than those of embryos reconstituted with MII oocytes ($96.7\%\;and\;23.9\%$, respectively, P<0.05). From the present result, it is suggested that domecolcine is useful for the enucleation of recipient oocytes in bovine NT procedures, and MII oocytes rather than MI oocytes are more appropriate for recipient cytoplasm. Although, the potential to develop into blastocysts of NT embryos produced by 1st PB-nonextruded and MI oocytes was very low, these oorytes could be used for NT.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.8 no.3
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• pp.185-195
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• 1975

The comparative studies Were conducted with respect to the artificial spawning early embryonic development, metamorphosis and growth of two species Meretrix lusoria and Cyclina sinensis collected from Inchon, Anmyon island and Buan areas from 1969 to 1974. The highest rate of artificial spawning of M. lusoria, which treated with a dilute ammoniun hydroxide(4/100-5/100N)-seawater solutions, was $25.0-33.3\%$, whereas in C. sinensis the rate of spawning was lower than that of M, lusoria under the similar experimental conditions$(12.5-19.0\%)$. However, the rate of artificial spawning of C. sinensis increased $40\%$ by repeated thermal stimulation. The rate of artificial fertilization of M. lusoria and C. sinensis showed highest value from those individuals which were treated with 1/1000N $NH_4OH$ solution. Their fertilized eggs, then, showed a normal development in the 1/1000N $NH_4OH$ solution. In the early embryonic development of M. lusoria and C. sinensis, the appearance of each of polar body, trochophore and D-shaped veliger were observed around 50min. 5-6 hours, and 23 hours after artificial fertilization respectively. The larval shell lengths of M. lusoria reached to $109,5{\pm}0.7\mu,\;144.6{\pm}1.3\mu$ and $208.0{\pm}0.0\mu$ around, 1, 11 and 20 days, after fertilization respectively. The larval shell lengths of C. sinensis reached to $110.5{\pm}0.6\mu,\;147.8{\pm}1.7\mu,\;and\;235.0{\pm}0.0\mu$ around 1, 10 ana 20 days, after fertilization respectively. The correlations of relative growth rate between the shell length(L) and sell height(H) found by the following simple formula from D-shaped veliger to metamorphosing stage. H=0.77L+6.82 for M. lusoria H=0.75L+8.50 for C. sinensis.