• Reproductive and Developmental Biology
• /
• v.32 no.1
• /
• pp.33-38
• /
• 2008

DNA methylation is involved in tissue-specific gene control and essential for normal embryo development Octamer-binding transcription factor 4 (Oct-4) is one of the most important transcription factors for early differentiation. This study was performed whether the bovine Oct-4 is tissue specific or developmental dependent epigenetic mark, we investigated transcripts and the methylation status of CpGs of 5'-promoter region of Oct-4 in bovine preimplantation embryos. Oct-4 transcripts were highly detected in morula and blastocyst, while they were present low levels in sperm and 2- to 8-cell stage embryos. These results suggest that de novo expression of Oct-4 initiates at morula stage of embryogenesis. Here we determined that there is a tissue-dependent differentially methylated region (T-DMR) in the 5'-promoter region of Oct-4. The methylation status of the Oct-4 T-DMR was distinctively different in the oocyte from that in the sperm and adult somatic tissues and changed from zygote to blastocyst stage, suggesting that active methylation and demethylation occur during preimplantation development. Based on these results, the 5'-promoter region of Oct-4 gene is target for DNA methylation and the methylation status changes variously during embryonic development in bovine.

• Research in Plant Disease
• /
• v.14 no.3
• /
• pp.176-181
• /
• 2008

This experiment was conducted to find the causes of ineffective seed disinfection methods such as rice seeds soaking in hot water and prochloraz EC solution when the rice seeds were severely infected by Bakanae disease. In case of rice seeds collected from severely diseased field by Bakanae disease, the pathogen as the forms of spores and mycelium were infected in plumule and inner and outer integument of embryo, aleurone layer, and pericarp layer. When the rice seeds were soaked in hot water, the appearances of spores and hypha on the outer pericarp layer were severely disordered, however those of inner region of outer integument and aleurone layer were shown normal. The membrane of hypha on the outer pericarp layer was destroyed within 24 hours, while some spores were healthy and germinated 7 days after soaking, when the rice seeds soaked 24 hours in 125 ppm prochloraz solution at $30^{\circ}C$. These results indicated that the seed disinfection methods were ineffective on the Bakanae disease severely infected rice seed because the hot water did not transmit the pericarp layer of rice seed and also prochloraz solution did not effectively destroy the spore of pathogen.

• Korean Journal of Medicinal Crop Science
• /
• v.6 no.4
• /
• pp.294-298
• /
• 1998

The factors affecting direct somatic embryogenesis from different parts of explant in liquid culture of Scrophularia buergeriana were investigated. Direct somatic embryogenesis was dependent on the explant tissues and stem was the most efficient explant. Rapid shoot development occurred on stem after 3-week culture but roots were not developed yet. Plantlets were not formed through somatic embryogenesis after 3-week culture of petiole. Though direct somatic embryo was not observed from leaf segment culture for 3 weeks, normal plantlets were developed after 8-week culture. BA played the main role for somatic embryogenesis in liquid culture and adding of either IAA or NAA caused rather adverse effects. Culture of stem segments in MS liquid medium with BA at 0.5 mg/ l or 0.1 mg/ l was proved to be the most efficient method for producing plantlets through direct somatic embryos.

• Korean Journal of Animal Reproduction
• /
• v.21 no.1
• /
• pp.47-52
• /
• 1997

The objective of this study was to investigate the in vitro / in vivo embryonic development after vitrification of mouse zygotes and the chrom osomal normality of delivered live young after embryo transfer. Mouse IVF zygotes were cryopreserved by vitrification using vitrification solution, EFS40 (40% ethylene glyc이, 30% Ficoll a and 0.3 M sucrose in phosphate buffer saline c containing 10% FBS ) . After mouse zygotes were exposed to EFS40 at 25"C for 30 sec., they were immediately plunged into LN$_2$. Vitrified thawed mouse zygotes were cultured upto bIastocysts in M16 for 4 days. The rates of in vitro development were 71.5% under this condition. Cultured blastocysts were transferred to recipients (3 day of pseudopregnant) on one or both uterus horns (6-8 embryos per a uterus horn). And all recipients were allowed to produce litters. The results obtained in these experiments were summarized as follows: The pregnancy rates and in vivo survival rates, live fetus rates, for vitrified zygotes (80.0, 39.6%) were not significantly difference in those of control zygotes (77.8%, 50.0%). Also, all of live-born young mice were chromosomally normal (n=40). This results suggested that proposed rapid vitrification procedures can be effectively use in 1-cell mouse zygotes cryopreservation.

• Reproductive and Developmental Biology
• /
• v.31 no.3
• /
• pp.193-198
• /
• 2007

The present study was examined the expression patterns of cdx2 gone, n lineage marker, in the mouse and bovine developmental stage embryos and whether one blastomere of two- and/or four-cell bovine embryos develop to specific lineage (ICM or TE) of blastocyst by injection of Texas red conjugated dextran as a lineage tracer. It was also investigated the allocation of ICM and n cells in bovine blastocysts derived from one blastomere of two-and/or four-cell stage embryos. Firstly, it was observed that expression of cdx2 appeared symmetric and asymmetric distribution at the two-cell stage mouse embryos. from four-cell to morula stage mouse embryos, the expression of cdx2 gene was observed in almost all blastomeres. In case of bovine embryos, localization of cdx2 was similar to pattern of mouse embryos. The Dextran-labeled blastomere of two- and/or four-cell embryos contributed to both ICM and TE cells in bovine blastocysts. And also, it was confirmed that a single blastomere derived from two-cell stage bovine embryos could develop to the normal blastocyst with both ICM and TE cells. These results show that two-and/or four-cell stage is not the specific stage to determine the cell rate for ICM and TE, and which is not correlated with the expression of cdx2 gene.

• Clinical and Experimental Reproductive Medicine
• /
• v.15 no.2
• /
• pp.135-147
• /
• 1988

To compare the stimulation effect of the ratio in follicle stimulating hormone and luteinizing hormone in induction of multiple follicular growth, the serum $E_2$ level, the diameter of follicle, number of aspirated follicles and cleavage rate of in vitro fertilized preovulatory oocytes as well as the pregnancy rate were evaluated. Forty one patients with irreparable tubal disease were stimulated by hMG(n=24) or FSH/hMG(n=17) for the purpose of in vitro fertilization and embryo transfer. The following results were obtained. 1. Serum estradiol($E_2$) levels on the day of hCG administration were $921.0{\pm}353.3\;pg/ml$ in hMG group and $1272.9{\pm}1060.6\;pg/ml$ in FSH/hMG group. The serum $E_2$ value of hMG group was significantly lower than that of FSH/hMG group. 2. The diameter of leading follicle by ultrasonogram on the day of hCG administration were $16.2{\pm}2.0\;mm$ in hMG group and $16.2{\pm}2.6\;mm$ in FSH/hMG group. No significant difference of follicle diameter between two groups was demonstrated. 3. The number of follicles with diameter above 10 mm by sonogram on the day of hCG injection were $3.91{\pm}2.32$ in hMG group and $6.52{\pm}3.86$ in FSH/hMG group. There was significant difference of number of follicles between two groups, (p< 0.01). 4. The number of oocytes found per patient at aspiration were $2.59{\pm}1.00$ in hMG group and 3. $76{\pm}2.31$ in FSH/hMG group. There was significant difference of number of aspirated oocytes between two groups. (p< 0.05). 5. The detection rate of preovulatory oocyte at aspiration were 68.4%(39/57) in hMG group (n=22) and 77.6%(38/49) in FSH/hMG group (n=13). 6. The cleavage rate of preovulatory oocyte at 44 hours after insemination were 74.4%(29/39) in hMG group(n=22) and 81.6%(31/38) in FSH/hMG group (n=13). When only hMG was used, one pregnancy was established in 15 patients to whom 29 zygotes were transferred. And a full term normal female baby was delivered by elective cesarean section. In the FSH/hMG group, five pregnancies out of 9 transferred patients were confirmed by serum ${\beta}-hCG$. Two pregnancies were spontaneously aborted before the 6th week of pregnancy. One patient aborted her baby at the 18th week of pregnancy because of incompetent internal os of the cervix. Two patients delivered two full term babies by elective cesarean section. From the above findings, paralell with the increase in the ratio of exogenous follicle stimulating hormone to luteinizing hormone, an increase in oocyte recovery was observed as well as an improvements in pregnancy rate. It was concluded that FSH enrichment early in the follicular phase had a beneficial effect in the controlled ovarian hyperstimulation.

• The Korean Journal of Zoology
• /
• v.38 no.4
• /
• pp.550-556
• /
• 1995

The c-myc proto-oncogene is Involved In the control of normal cell proliferation and differentiation of many cell lineages. Although it has heen suggested that c-myc may play an important role in the mammalian early development, it Is unclear whether the embryonic c-myc mRNA is originated from zygotic gene expression or stored maternal message. Thus, we have construded expression vectors, In which the 5, flanking sequences including c-myc promoter region and a large non-coding exon I are fused 'sith E. coli lacZ gene that encedes $\beta$-galactosldase as a reporter. As c-myc exon I contains a modulatory sequence, we designed t, vo types of vectors (pcmyc.Gall and pcmyc-Ga12) to examine the role of exon I in c-myc expression. The former contains the complete exon I and the later has a deletion in 40 bp of modulator sequence located In the exon I of c-myc These vectors were microInjected into fertilized one-cell embryos and $\beta$-galactosidase activity was examined by X-gal staining during early embryogenesis. $\beta$-galactosidase activity derived from c-myc promoter was decreased at two-cell stage. The expression level directed by pcmyc- Ga12 was similar to that of pcmyc-Gal1, indicating that the medulatory sequence in exon I may not be Involved at least In the regulation of embryonic c-myc expression. In summary, the present study indicates that the c-myc promoter is functional at the early stage embryo, and the regulation of c-myc expression is under the control of "zygotic" clock of preimplantation mouse embryos.e embryos.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.31 no.1
• /
• pp.114-120
• /
• 1998

The spawning behavior, development of eggs and larvae of the Korea freshwater goby, Rhinogobius brunneus (Temminck et Schlegel) were studied. The eggs were spawned as a one-layer mass, hanging from the underside of a small pebble, and guarded by one male. The eggs were elliptic, about 1.48 mm in length and 0.65 mm in breadth, with a round top and a somewhat flat base with glutinous fibers. Hatching in the indoor tank with $17.0^{\circ}C$ in mean water temperature started from the 146 hours after fertilization. In the late developing stages, the embryo moved and freely changed its head to face the free tip of the chorion (normal egg) or to the basal end with a boundle of adhesive filaments (agrippa egg). Newly-hatched larvae were $3.10\~3.30$ mm in total length (mean: 3.22 mm), and mouth and anus were not yet open. Melanophores were present on the air- bladder, around the anus, and on the ventral part of the caudal region. The larva $3\~4$ days old transformed to postlarval stage, and they were $3.30\~3.85$ mm in total length (mean: 3.60 mm). As yolk sac and oil globules werw nearly absorbed, mouth and anus were open, and they fed rotifers actively. In $20\~22$ days after hatching the larvae grew to 5.85 mm in 71, and the caudal notochord flex at $45^{\circ}$.

• Korean Journal of Plant Resources
• /
• v.35 no.2
• /
• pp.183-191
• /
• 2022

Ginseng (Panax ginseng C.A. Meyer) is a perennial plant and propagates by seeds, and those need after-ripening for germination. To be ready for climate change and to ensure a stable seed supply, the technique for storing seeds in short-term and long-term in large quantities is required. In this study, dehisced ginseng seeds from two locations, batch #1 and batch #2, were stored at -3.5℃ with different moisture content, and after 3, 15, and 27 months of storage, the percentage of radicle emergence and shoot emergence were measured. After 3 months, radicle emergence and shoot emergence were normal only when the seed moisture content was more than 35%, and overall, germination was higher in batch #2 than in batch #1. After 15 months, the partially dehydrated seeds, with a moisture content between 45 to 54%, showed the highest germination rates, and most of the undried seeds were spoiled and failed to germinate. Seeds with moisture content lower than 25% had poor germination, too. The partially dehydrated seeds also succeeded in germination and growth in the soil after 15 months of storage, but deteriorated after one more 1 year, too. In summary, ginseng seeds look like have temperate recalcitrant seed characteristics, and partial dehydration allows extension of seed longevity.

• Animal Bioscience
• /
• v.36 no.12
• /
• pp.1775-1784
• /
• 2023

Objective: The aim of this study was to reveal the role and regulatory mechanism of miR-188-5p in the proliferation and differentiation of goat muscle satellite cells. Methods: Goat skeletal muscle satellite cells isolated in the pre-laboratory were used as the test material. First, the expression of miR-188-5p in goat muscle tissues at different developmental stages was detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR). In addition, miR-188-5p was transfected into goat skeletal muscle satellite cells by constructing mimics and inhibitors of miR-188-5p, respectively. The changes of differentiation marker gene expression were detected by qPCR method. Results: It was highly expressed in adult goat latissimus dorsi and leg muscles, goat fetal skeletal muscle, and at the differentiation stage of muscle satellite cells. Overexpression and interference of miR-188-5p showed that miR-188-5p inhibited the proliferation and promoted the differentiation of goat muscle satellite cells. Target gene prediction and dual luciferase assays showed that miR-188-5p could target the 3'untranslated region of the calcium/calmodulin dependent protein kinase II beta (CAMK2B) gene and inhibit luciferase activity. Further functional studies revealed that CAMK2B promoted the proliferation and inhibited the differentiation of goat muscle satellite cells, whereas si-CAMK2B restored the function of miR-188-5p inhibitor. Conclusion: These results suggest that miR-188-5p inhibits the proliferation and promotes the differentiation of goat muscle satellite cells by targeting CAMK2B. This study will provide a theoretical reference for future studies on the molecular mechanisms of skeletal muscle development in goats.