• Development and Reproduction
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• v.7 no.2
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• pp.95-103
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• 2003

Intracellular calcium is an important physiological factor in most cells, and ruthenium red and ryanodine play an important role as calcium modulators. Ruthenium red inhibits calcium-induced calcium release(CICR) from the intracellular calcium store. Ryanodine activates calcium release through ryanodine channel. The present experiment was performed to investigate the effects of two modulators on calcium ion metabolism and to determine their dose-dependency in oocyte and early embryo of mouse. Intracellular calcium ion concentration was measured in realtime by using confocal laser scanning microscope(CLSM) after loading of Fluo-3/AM in mouse oocytes and early embryos. Ruthenium red decreased intracellular calcium ion concentration in oocytes and early embryos at its high concentration(30, 300 $\mu$M). Ryanodine increased intracellular calcium ion concentration in oocytes and early embryos in low concentration(0.01 $\mu$M) but decreased that at higher concentrations(1, 10 $\mu$M). These results indicate that two modulators affected calcium ion metabolism in oocyte and early embryo of mouse, and their dose-dependency was different from somatic cell including myocytes.

• Clinical and Experimental Reproductive Medicine
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• v.23 no.3
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• pp.293-301
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• 1996

In the present study, mouse follicular oocytes and 2-cell embryos(late -zygote stage embryos included) were cultured on the electric pad for electric stimulation in the culture incubator. In addition, follicular oocytes and embryos were tested for maturation and development under higher temperature condition($39^{\circ}C$).Mouse follicular oocyte maturation were not affected by anion electric stimulation and there is no significant difference in GBVD and MI between the control and experiment group after 4hr culture. In the embryo culture, it was found that more morula and blastocyst were found in the electric stimulation group rather than the control(96hr). This may seem to be caused with cytoplasmic $Ca^{2+}$ transient rise by electric stimulation(anion pad). On the other hand higher temperature incubation ($39^{\circ}C$) on the anion pad caused all the embryos degenerated within $12h{\sim}24hr$ culture. This was quite different from large animal embryos(bovine, pig, sheep), in which beneficial effect of high temperature incubation for oocyte maturation and embryo development were found.

• Korean Journal of Agricultural Science
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• v.44 no.3
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• pp.309-317
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• 2017

Melatonin (N-acetyl-5-methoxytryptamine) is an indole synthesized from tryptophan by the pineal gland in animal. The major function of melatonin is to modulate circadian and circannual rhythms in photoperiodic mammals. Importantly, however, melatonin is also a free radical scavenger, anti-oxidant, and anti-apoptotic agent. Recently, the beneficial effects of melatonin on oocyte maturation and embryonic development in vitro have been reported in many species such as pig, cattle, sheep, mouse, and human. In this review, we will discuss recent studies about the role of melatonin in the production of porcine and bovine oocytes and embryos in vitro in order to provide useful information of melatonin in oocyte maturation and embryo culture in vitro.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2006.10a
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• pp.111-112
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• 2006

• Journal of Embryo Transfer
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• v.16 no.3
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• pp.239-243
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• 2001

Selection of oocyte cryopreservation method is a prerequisite factor for developing an effective bank system. Compared with slow freezing method, the vitrification has various advantages such as avoiding intracellular ice crustal formation. In our previous, we attempted to employ a vitrification method using ethylene glycol and an electron microscope grid for cryopreservation of mouse oocytes. However, A high incidence of spindle and chromosome abnormalities was detected in thawed oocytes after vitrification. We examined whether the addition of a cystoskeleton stabilizer Taxol $^{TM}$, to the vitrification solution could promote the post-thawed survival and subsequent development of stored oocytes. More oocytes developed to the 4-cell (44.7% vs. 69.7%), 8-cell (31.8% vs. 64.2%), morula (24.7% vs. 54.3%), and blastocyst (20.3% vs. 49.2%) stages after the addition of Taxol$^{TM}$ to the cryoprotectant than after no addition. 21 and 26 mouse pups were born after transfer of blastocyst derived from oocytes vitrified without and with Taxol. The addition of Taxol to vitrification solution greatly promoted post-thaw preimplantation development of ICR morose oocytes.tes.

• Journal of Embryo Transfer
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• v.33 no.2
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• pp.69-73
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• 2018

This study was conducted to examine the influences of two human chorion gonadotrophins (hCGs) being injected into young or aged (45- to 65-week old) outbred (ICR) mice on developmental capacity of oocytes retrieved. In vitro-culture and parthenogenetic activation of oocytes retrieved were employed for the assessment. Superovulation was determined as being induced when more than 25 oocytes were retrieved. No aged mice were superovulated, while in contrast, 67-100% were superovulated in the 6- to 8-week-old (young) mice. In the aged, hCG injection yielded better retrieval (5 vs. 13 to 14.8 oocytes/mouse). Overall, no significant difference between two hCGs was detected but between the young and aged, significant differences in maturational arrest (0% vs. 39% MI arrest and 46% vs. 15% degeneration) and developmental capacity (24% vs. 46% 8-cell embryo development) were detected. In conclusion, hCG injection contributes to increasing oocyte retrieval from aged outbred mice, but the kinds of gonadotrophin influenced the efficiency of hyperstimulation induction in specific ages.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.113-113
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• 2002

The stimulatory effect of EGF and FSH on oocyte maturation have been reported in various mammalian species. And some reports presented FSH enhanced the effect of EGF on oocyte maturation. But, the interaction between EGF and FSH on nuclear maturation of mammalian oocytes is not fully understood. We observed the effect of EGF and FSH on nuclear maturation during in vitro maturation of mouse oocytes. Also, we examined the interaction between EGF and FSH on nuclear maturation of mouse oocytes using the EGFR inhibitor or FSH inhibitor. Germinal vesicle (GV) stage oocytes were obtained from 3-4weeks PMSG primed BCFI hybrid mice and cultured in TCM-199 medium with 0.4%PVP supplemented with/without EGF (1ng/ml), FSH (1ug/ml), EGFR specific tyrosine kinase inhibitors: Tyrphostin AG 1478 (500nM), MAP kinase kinase inhibitor : U0126 (20uM) or PD 98059 (100uM) for 14-l5hr. Rapid staining method were used for the assessment of nuclear maturation. Nuclear maturation rates of EGF indjor FSH-treated group were significantly higher than those of control group. Treatment of EGFR inhibitor significantly block the nuclear maturation of GV oocyte in EGF-treated group, but it did not block those of GV oocyte in FSH-treated or FSH and EGF-treated group. Treatment of FSH inhibitor(U0126, PD98059) significantly block the nuclear maturation of EGF-treated group, FSH-treated and FSH and EGF-treated group. These results show that EGF has a stimulatory effect as well as different action pathway with FSH on in-vitro maturation of mouse oocyte in vitro. Therefore, further studies will be needed to find the signaling pathway of EGF associated with nuclear maturation.

• Clinical and Experimental Reproductive Medicine
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• v.21 no.3
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• pp.325-330
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• 1994

The study has been carried out in order to evaluate the effects of embryonic stage, and cryopreservation method on the rates of viability and development of the cryopreserved mouse early embryos. The results were as following:In the treatment steps of cryoprotectant, for the fertilized oocyte with pronucleus(PN), 2-step was better than the others. And for the other embryos, 4-step was better than 2- or 3-step. In respect to the embryonic stage, as the embryos developed from fertilized oocytes to 8-cell embryos, the rates of viability and development were increased higher. Therefore, 8-cell embryo was better stage than the others. In respect to the kind of cryoprotectants, PROH was better than DMSO for the fertilized oocyte, as a cryoprotectant. DMSO, for the 2-cell embryos and PROH and DMSO for the 4- and 8-cell embryos were suitable for cryopreservation.

• Journal of Animal Science and Technology
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• v.65 no.4
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• pp.767-778
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• 2023

The aim of the research is to identify that porcine oocytes can function as recipients for interspecies cloning and have the ability to develop to blastocysts. Furthermore each mitochondrial DNA (mtDNA) in interspecises cloned embryos was analyzed. For the study, mouse-porcine and porcine-porcine cloned embryos were produced with mouse fetal fibroblasts (MFF) and porcine fetal fibroblasts (PFF), respectively, introduced as donor cells into enucleated porcine oocytes. The developmental rate and cell numbers of blastocysts between intraspecies porcine-porcine and interspecies mouse-porcine cloned embryos were compared and real-time polymerase chain reaction (PCR) was performed for the estimate of mouse and porcine mtDNA copy number in mouse-porcine cloned embryos at different stages.There was no significant difference in the developmental rate or total blastocyst number between mouse-porcine cloned embryos and porcine-porcine cloned embryos (11.1 ± 0.9%, 25 ± 3.5 vs. 10.1 ± 1.2%, 24 ± 6.3). In mouse-porcine reconstructed embryos, the copy numbers of mouse somatic cell-derived mtDNA decreased between the 1-cell and blastocyst stages, whereas the copy number of porcine oocyte-derived mtDNA significantly increased during this period, as assessed by real-time PCR analysis. In our real-time PCR analysis, we improved the standard curve construction-based method to analyze the level of mtDNA between mouse donor cells and porcine oocytes using the copy number of mouse beta-actin DNA as a standard. Our findings suggest that mouse-porcine cloned embryos have the ability to develop to blastocysts in vitro and exhibit mitochondrial heteroplasmy from the 1-cell to blastocyst stages and the mouse-derived mitochondria can be gradually replaced with those of the porcine oocyte in the early developmental stages of mouse-porcine cloned embryos.

• Journal of Embryo Transfer
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• v.16 no.3
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• pp.233-237
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• 2001

To develop an effective vitrification method, we examined the use of a conventional straw as vessel fur vitrification of mouse oocytes, and to compare the post-thaw survival and chromosome configuration of these oocytes with those vitrified in grids. Intact cumulus-enclosed oocytes were vitrified with DPBS with 5.5 M ethylene glycol and 1.0 M sucrose, and loaded into straws and onto eletron microscopic copper grid fur storing in liquid nitrogen. Intact vitrified and thawed oocytes were karyotying for chromosome. The rates of post-thawed survival were 88.5% in vitrified oocytes with straws, and 83% in vitrified ooctyes with grids. Vitrified and thawed oocytes with straws and grids were increased chromosomal abnormality (31.4% and 30.9%) compared with fresh oocytes (17.8%). The conventional straws can be used as vessel for vitrification to prevent of inflection in liquid nitrogen.