• Korean Journal of Ichthyology
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• v.32 no.4
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• pp.222-231
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• 2020

The egg development and early life history of Sharpbelly Hemiculter leucisculus were investigated. For the experiments, the mature adults were collected at the Lake Yedang in Korea. The eggs from the females were obtained by injecting 10 IU/g of human chorionic gonadotropin and inseminated by wet method in the laboratory. The fertilized eggs were 0.97±0.02 mm (0.9~1.0 mm n=30) in diameter. The embryo began to hatch about 32 hrs after fertilization under water temperature of 22±1℃. The newly-hatched larvae were 3.0±0.2 mm (2.6~3.4 mm, n=15) in total length, and they haven't Melanophore. Six days after hatching, the Preflexion larva were 5.7±0.1 mm (5.4~5.8 mm, n=15) in total length, and they began to eat a Rotifer. 17 days after hatching, the Flexion larva were 6.8±0.2 mm (6.5~7.0 mm, n=15) in total length, and a gas bladder develop above the intestine. 30 days after hatching, the Postflexion larva were 8.8±0.7 mm (7.9~10.3 mm, n=15) in total length, three dorsal fin rays began to develop in the membrane fins. 50 days after hatching, the Juvenile were 20.8±0.8 mm (18.8~24.6 mm, n=15) in total length, and all their fin-rays were formed.

• Reproductive and Developmental Biology
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• v.29 no.1
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• pp.1-7
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• 2005

Telomeres consisting of (TTAGGG)n tandem repeat DNA sequences and associated proteins are essential for chromosome stability and related with cell senescence, apoptosis and cancer. The telomerase is a ribonucleoprotein which act as a template for the synthesis of telomeric DNA. This study was carried out to identify the distribution of telomeres on mouse chromosomes and also to analyze the amount of telomeres and telomerase activity of mouse embryos at early embryonic stages. Germ cells and early embryos from 1 cell to blastocyst stage were analyzed. The amount of telomeres was analyzed by quantitative fluorescence in situ hybridization technique(Q-FISH) using a human telomeric DNA probe, and telomerase activity was measured by telomeric repeat amplification protocol assay(TRAP). In results, the telomeres on mouse chromosomes were distributed at the ends of all autosomes and sex chromosomes. Although the quantity of telomeres varied among chromosomes, most of chromosomes had higher amount in q-arm telomeres than in p-arm telomeres. The results of Q-FISH indicated that the relative amount of telomeres of mouse embryos in each embryonic stage was approximately the same except the higher amount in blastocysts. Using TRAP assay on mouse embryos, telomerase activity was detected in all preimplantation stages from mature oocytes to blastocysts. Especially the telomerase activity was significantly increased at the morula and blastocyst stage. In conclusion, there may be a close association between the amount of telomeres and telomerase activity in early embryonic stages, and analysis of telomere quantity and telomerase activity on early development will be helpful for the investigation of embryogenesis and embryonic cell differentiation in mice.

• The Korean Journal of Pesticide Science
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• v.11 no.4
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• pp.254-260
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• 2007

In order to evaluate the toxic effects of butachlor, a herbicide widely used for control of weeds in paddy field, on medaka (Oryzias latipes), acute toxicity tests for five developmental stages and early life stage toxicity test of were conducted. As the results of acute toxicity test, $96h-LC_{50}s$ for 1 day, 1 week, 2 weeks, 2 months and 5 months after hatching of O. latipes were 0.68, 0.52, 0.38, 1.09 and $0.45\;mg\;L^{-1}$, respectively. This indicated that the most sensitive stage was 2 weeks after hatching. The early life stage toxicity test showed that no statistically significant hatching period and hatching success of embryo was observed at all concentrations of butaclor. However, 0.05 and $0.1\;mg\;L^{-1}$ of butachlor showed statistically significant post hatching survival with p<0.1. Abnormalities of larva were 2.1, 2.3 and 10% at 0.025, 0.05 and $0.1\;mg\;L^{-1}$ of concentration, respectively. They showed abnormal vertebral axis, craniofacial alteration and retarded yolk-sac resorption. The total length and weight were decreased depending on butachlor concentration the end of test. Weight of larva was showed more sensitive toxic indicator than total length. The toxicological responses of O. latipes to butachlor expressed as LOEC(lowest observed effect concentration), NOEC(no observed effect concentration) and MATC(maximum acceptable toxicant concentration) values were 0.025, 0.013 and $0.018\;mg\;L^{-1}$, respectively.

• Clinical and Experimental Reproductive Medicine
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• v.23 no.3
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• pp.319-326
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• 1996

The objective of this study was to investigate correlation between the morphology by microscopic assessments of surplus blastocysts produced in human IVF program and their cell number obtained by differential labelling method. For these experiments, 76 surplus human blastocysts were obtained from 36 patients on day 5 after IVF, the embryos were classified to early (ErB), early expanding (EEB), middle expanding (MEB), expanded blastocyst (EdB) according to their blastocoel expansion and zona thickness. When the ovum size and zona thickness of the classified blastocysts were measured using micrometer, although the embryos were produced in the same culture condition, there were significant variances in ovum size ($148.8 217.6{\mu}m$) and zona thickness ($1.2-14.4{\mu}m$). Total blastomere cell number counted after hoechst staining was increased by two to three fold during the transition period from ErB ($39.1{\pm}3.6$) to EdB ($(89.6{\pm}3.3)$) stage on day 5 after IVF. ICM ($11.9{\pm}1.8-22.2{\pm}4.3$) and TE ($24.5{\pm}3.6-70.0{\pm}7.7$) cell numbers using differential labelling were also showed the increased pattern according to the developmental level. Especially, EdB which showed poor ICM morphologically also indicated the low ICM cell number after differential labelling. This demonstrated that there is good correlation between the morphological assessment and the cell number. The count of ICM and TE nuclei using differential labelling can be used as an important criterion, if it is accompanied with morphological assessments, in selecting the better embryos for improving the pregnancy rates in human blastocyst transfer program.

• Clinical and Experimental Reproductive Medicine
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• v.24 no.3
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• pp.347-353
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• 1997

This study was to test whether in vitro/in vivo survival of vitrified mouse blastocysts was influenced by culture conditions and ET method. Mouse blastocysts were obtained from in vitro fertilization and cultured for 4 days in M16 medium, and they were vitrified in EFS40 which contained 40% ethlyene glycol, 18% Ficoll and 0.5 mol sucrose in PBS. In experiment I, in vitro and in vivo survival rate of these embryos were evaluated in different culture condition after thawing. When thawed embryos were cultured in M16 medium as a control, m-CR1 medium contained 20 amino acids (2% BME amino acis and 1% MEM non-essential amino acids solution) and 4 mg/ml BSA and cumulus monolayer cell co-cultured condition in mCR1 medium (10% FBS), their in vitro survival at 24 hr after thawing was not affected by culture condition (75.6, 83.1, 82.4%). However, in vivo survival rates of implantation in m-CR1 medium (80.4%) were significantly higher than those of M16 medium (51.2%), co-culture (57.1%) condition, although there was no difference in live fetuses rates on day 15 gestation (39.0, 49.0, 38.1%). In experiment II, the in vivo development potential of embryos by ET methods was examined. When blastocysts were transferred to the day 2, 3 pseudopregnant recipient without culture soon after thawing, no pregnant recipient was obtained on the day 2 pseudopregnancy, and 50% of pregnancy rates and 15.4% of live fetus rates were obtained on the day 3 pseudopregnant recipients. These results were significantly lower than those of transferred group (day 3 pseudopregnant recipients) after culture for 16 hr post thawing (73.5, 57.1%) (p<0.05). In experiment III, to elevate usability of delayed embryos in vitro/in vivo survival of vitrified embryos (day 4 early, day 5 early and expanding blastocyst) were examined. in vivo survival rates (live fetus, total implantation) were higher in day 4 early blastocysts (33.3, 66.7%) than in day 5 expanding blastocysts (29.0, 38.7%), although the highest in vitro survival rates were obtained in the day 5 expanding brastocysts (78.3%). Therefore, these results suggest that the in vitro/in vivo survival rates of vitrified embryos could be improve by the culture condition and ET method and that the in vivo development rates of delayed embryos were decreased with longer culture duration in vitro. It means that more effective cryopreservation was obtained in day 4 early blastocysts than in day 5 expanding blastocysts.

• Journal of the korean academy of Pediatric Dentistry
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• v.28 no.4
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• pp.709-727
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• 2001

The pattern of programmed cell death(PCD) has been examined during the early developmental period of development in mouse embryos, from embryonic day 4.5(E4.5) to E11.5 Embryos from Balb/c breedings were harvested at various embryonic stages between E4.5 and El1.5. Cell death was analysed by in situ terminal deoxynucleotidyl transferase mediated dUTP nick end labeling(TUNEL) staining in tissue sections and whole embryos. At the blastocyst stage(E4.5), a very few apoptotic cells were found in the inner cell mass of the blastocyst. In the early egg cylinder stage(35.0-5.5), a few apoptotic cells were detected in the embryonic ectoderm, the embryonic endoerm and the proamniotic cavity. In the advanced egg cylinder stage(E5.5-6.5), TUNEL-posifive cells were observed in the extra-embryonic ectoderm and extra-embryonic endoderm as well as in the embryonic ectoderm, embryonic visceral endoderm and proamniotic cavity. In the streak stage(E6.75-7.75), many TUNEL-positive cells were found in the ectoplacental cone. In contrast, only very few apoptotic cells were found in the chorion and extra-embryonic endoderm in extra-embryonic regions. In intra-embryonic region, a few apoptotic cells were randomly found in the embryonic ectoderm, mesoderm and visceral endoderm. At the early somitogenesis stage(E8.0-8.5), most apoptotic cells were observed in the most cranial portion of neural fold (neural ectoderm and adjacent ectoderm). At the mid somitogenesis stage(39.0-9.5), the otic placode first showed TUNEL-positive at this stage. Small number of TUNEL-positive cells were also first seen around optic placode and branchial arches. Three streams of TUNEL-positive cells were clearly seen in the cranial region at 59.5-9.75. At E10.5, apoptotic cells were localized in the developing eye, the junctional portion of medial nasal, lateral nasal and maxillary processes, the lateral portion of branchial arches, the junction of bilateral mandibular processes, and apical ectodermal ridges of limb buds. At E11.5, apoptotic cells were noticeably decreased in most area, except the developing limbs and several somites in the tail region. In this study, the global temporospatial pattern of PCD throughout early development of mouse embryos was discussed. It may provide the basis for further studies on its role in the morphogenesis of the embryo.

• The Korean Journal of Malacology
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• v.23 no.2
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• pp.189-198
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• 2007

The reproductive cycle, egg capsules in the egg-mass, first sexual maturity, and sex ratio of the bladder moon, Glossaulax didyma ($R\ddot{o}ding$) were investigated. The gastropods collected from the intertidal zone of Biin Bay, Seocheon, Korea were studied by using histological analysis and morphometric data. The gonadosomatic index (GSI) of females and males began to increase in March and reached maximum in May. Then their values sharply decreased from late in May to August due to spawning. The condition index (CI) began to increase in February and reached maximum in May, then gradually declined in the spawning period. The CI calculated for determination of the spawning period was coincided with changes in the GSI and gonadal phases. Spawning occurred between late in May to August in females and early in May to August in males. Spawning peak was observed between July and August when the seawater temperature rose to 19 $^{\circ}C$. Reproductive cycle with the gonadal development phases of this species can be divided into five successive stages in females and four in males: in females, early active stage (December to February), late active stage (February to March), ripe stage (April recovery stage (August to November); in males, active stage (December to March), ripe stage (March to July), copulation stage (early May to August), and recovery stage (August to January). Fully matured oocytes were approximately 250-270 ${\mu}m$ in size. The egg-mass was a hat in shape, and a number of egg capsules were found in an egg-mass. An egg capsule was 0.53-0.57 mm in size. An embryo (veliger larva) hatched from an egg capsule. Percentage of first sexual maturity in females and males were over 50% for individuals that are 40.1-45.0 mm in shell radius, and 100% for those that are over 45.1 mm. The sex ratio of female to male was significantly different from 1:1 $(x^2\;=\;57.22,\;p\;<\;0.05)$.

• Journal of Embryo Transfer
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• v.22 no.2
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• pp.121-126
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• 2007

The objective of this study was carried out to examine the selection effects of in vitro matured porcine follicular oocytes with polar body extrusion and early cleavage as non-invasive marker to know the developmental competence in advance. The porcine oocytes matured for 48 h were examined the polar body extrusion. The examined oocytes were matured for additional $16{\sim}18h$ and activated with 7% ethanol and cultured in $5{\mu}g/ml$ cytochalasin B for 5 h for diploid formation. The treated oocytes were cultured and examined the cleavage after 48 h and continued culturing for 5 days. The oocytes of 21.9% were discarded in morphological selection and 32.1% oocytes were discarded by failure of first polar body extrusion. The selected oocytes were matured and activated and then after 48 h the cleavage rates were examined. In morphologically selected oocytes, 15.8% oocytes were not cleaved and 52.6% oocytes were normally cleaved and 31.6% oocytes were hyper-cleaved over 8-cell stage. However in the first polar body extruded oocytes, 7.1% oocytes were not cleaved and 73.1% oocytes were normally cleaved and 19.8% oocytes were hyper-cleaved. The morphologically selected embryos that not cleavage-selected were developed in 16.7% up to blastocyst and the morphologically selected and cleavage-selected embryos were developed in 31.7%. The polar body extruded oocytes that were not carried out cleavage selection were developed in 39.0% and the polar body extruded and cleavage-selected embryos were developed 49.0%. The first cleavage timing was examined with 12 h interval after activation. In $0{\sim}12,\;12{\sim}24,\;24{\sim}36,\;and\;36{\sim}48h$ intervals, 4.1%, 68.6%, 19.1%, and 2.3% oocytes were cleaved and 5.9% oocytes were not cleaved until 48 after activation. The cleaved oocytes in each interval were cultured and developed upto blastocyst with 0, 39.1, 9.5, and 0%, respectively. This results suggests that polar body extruded and cleaved at $12{\sim}36h$ embryo has higher developmental potential than the others.

• Journal of Embryo Transfer
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• v.20 no.3
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• pp.279-287
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• 2005

This study was carried out to investigate resumption of ovarian cyclicity and effect of BCS on estrous response by treatment of $PGF_2a+PGF_2a+CIDR$ program on day 40 postpartum in lactating dairy cow. First $PGF_2a$ was given on day 40 postpartum, second $PGF_2a$ was given 14 days apart to cows not-responded to 1st $PGF_2a$ and then CIDR was inserted for 7 days after 5 days in cows not-responded to 2nd $PGF_2a$. The $42.9\%$ of the cows showed more than 1 ng/mL milk progesterone concentration within 10 to 30 days postpartum. About $19\%$ of the cows exhibited more than 1 ng/mL milk pro-gesterone concentration between 31 to 50 days postpartum. However $38.1\%$ of the cows have not shown more than 1 ng/mL milk progesterone up to 50 days postpartum. Estrous response to the treatment of 1 st $PGF_2a$ and 2nd $PGF_2a$ was $47.5\%$ and $52.4\%$, respectively. Combination of 1 st $PGF_2a$ and 2nd $PGF_2a$ was $75\%$ and combination of 1st $PGF_2a$+2nd $PGF_2a$+CIDR was $87.5\%$. Estrous response to the treatment of $PGF_2a+PGF_2a$ program was $61.5\%$ in cows with less than 2.50 BCS and $81.5\%$ in cows with 2.75${\~}$3.50 BCS. Estrous response to the treatment of CIDR was $40\%$ in cows with less than 2.50 BCS and $80\%$ in cows with 2.75${\~}$3.50 BCS. Estrous response to the treatment of PPC on day 40 postpartum was $76.9\%$ in cows with less than 2.50 BCS and $96.3\%$ in cows with 2.75${\~}$3.50 BCS.

• Journal of Embryo Transfer
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• v.19 no.2
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• pp.89-99
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• 2004

Stretch-activated channels (SACs) responds to membrane stress with changes in open probability (Po). They play essential roles in regulation of cell volume and differentiation, vascular tone, and in hormonal secretion. SACs highly present in Xenopus oocytes and Ascidian oocytes are suggested to be involved in the regulation of pH and fluid transport to balance the osmotic pressure, but remain unclear in mammanlian oocytes. This study was investigated to find the presence of SACs in hamster oocytes and to examine their electrophysiological properties. To infer a role of SAC in relation to the development of early stage, we followed up to the stage of two-cell zygote with patch clamp techniques. Single channels were elicited by negative pressure (lower than ­15 cm$H_2O$). Interestingly, SACs were dependent on permeable cations such as $Na^+$ or $K^+$. As permeable cation removed from both sides across the membrane, SAC activity completely disappeared. When permeable cations present only in intracellular compartment, outward currents appeared at positive potentials. In contrast to this, inward currents occurred only at the negative voltage when permeable cation absent in cell interior. These result suggests that SAC carry cations through the nonselective cation channel (NSC channel). Taken together, we found that stretch activated channels present in hamster oocyte and the channel may carry cations through NSC channels. This stretch activated-NSC channels may play physiological role(s) in oocyte growth, maturation, fertilization and embryogenesis in fertilized oocytes to two-cell zygotes of hamster.