• The Korean Journal of Zoology
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• v.23 no.4
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• pp.263-272
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• 1980

Polyacrylamide gel electrophoresis was used to investigate the patterns of LDH and MDH isozymes in the embryo and adult of amphibia; Rana nigromaculata, Rana plancyi chosenica and Hynobius leechii. Rana nigromaculata is considered to be heterozygous for the gene specifying the "B" subunit of LDH, and Hynobius leechii to be heterozygous for the gene specifying the "A" subunit of LDH. The LDH isozyme paatern of embryos of the above three species is characterized by a gradual increase in the activity of LDH-5 (muscular form)during development. Two or three molecular forms of MDH is present steadily from early embryos and in adult. Of the MDH isozymes, the more cathodic one (MDH-m) appears weakly in early developing stages, but increases slowly in the activity as the embryo develops.the embryo develops.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.12 no.8
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• pp.3541-3546
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• 2011

Autophagy is an evolutionarily conserved lysosomal pathway for degrading cytoplasmic proteins, macromolecules, and organelles in addition to recycling protein and ATP synthesis. Although autophagy is very important during embryogenesis, the mechanism underlying the dynamic development during this process remains largely unknown. In order to obtain insights into autophagy in early embryo development, we analyzed gene expression levels of autophagy-related genes (ATGs) in mouse embryos developing in vitro. Using real time RT-PCR technique, ATGs including Atg2a, Atg3, Atg4b, Atg5, Atg6, Atg7, Atg9a, and Wipi3, as maternal transcripts, were only up-regulated in 1-cell embryo stage before zygotic genomic activation (ZGA), and then expression decreased from 2-cell to blastocyst embryo stage. ATGs including Dram and Atg9b were expressed abundantly in 1-cell embryo state and in blastocyst embryo stage, athough Atg8 and Ulk1 were constantly expressed during preimplantation stage. However, Atg4d were only up-expressed from 4-cell to blastocyst stage. These results suggest that autophagy is related in mouse embryo, which possibly gives an important role for early development.

• Reproductive and Developmental Biology
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• v.37 no.2
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• pp.65-73
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• 2013

The objective of this study was to examine the effects of high concentrations of glucose on porcine parthenotes developing in vitro. Addition of 55 mM glucose to the culture medium of embryos at the four-cell-stage significantly inhibited blastocyst formation, resulting in fewer cells in blastocyst-stage embryos and increased levels of apoptosis and autophagy compared to control. Quantitative reverse transcriptase (RT) PCR analysis revealed that the expression of pro-apoptotic genes (Caspase 3, Bax and Bak) and autophagy genes (Atg6 and Atg8/Lc3) were increased significantly by the addition of 55 mM glucose to the culture medium compared to control. MitoTracker Green fluorescence revealed a decrease in the overall mitochondrial mass compared to control. However, the addition of 55 mM glucose had no effect on mRNA expression of the nuclear DNA-encoded mitochondrial-related genes, cytochrome oxidase (Cox) 5a, Cox5b and Cox6b1. These results suggest that hyperglycemia reduced the mitochondrial content of porcine embryos developing in vitro and that this may hinder embryonic development to the blastocyst stage and embryo quality by increasing apoptosis and autophagy in these embryos.

• Korean Journal of Poultry Science
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• v.28 no.2
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• pp.155-163
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• 2001

Gene and cell transfer technique will serve as a powerful tool for the genetic improvement of the poultry and to yield useful products. For avian transgenesis, Japanese quail may serve as an excellent animal model because of its small body size and fast growth rate. Recent progress was described on the manipulation of quail embryos such as the introduction of foreign genes and cells, and the subsequent culturing of the manipulated embryos yielding hatchlings. Intraspecific donor-derived offspring have been available in quail, however, further investigation will be required to obtain interspecific offspring with the aim of rescuing endangered species. Trans genesis will also be useful for improving the profitability and quality of poultry stocks and for developing stocks with novel uses. Considerable progress should soon be made toward the production of transgenic poultry. The key feature of the procedure described here is that embryos are initially taken out from the shell for ease of manipulation and then placed back in culture in addition to various operations midway during culture.

• BMB Reports
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• v.49 no.10
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• pp.572-577
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• 2016

The short gastrulation (sog) shadow enhancer directs early and late sog expression in the neurogenic ectoderm and the ventral midline of the developing Drosophila embryo, respectively. Here, evidence is presented that the sog primary enhancer also has both activities, with the late enhancer activity dependent on the early activity. Computational analyses showed that the sog primary enhancer contains five Dorsal (Dl)-, four Zelda (Zld)-, three Bicoid (Bcd)-, and no Single-minded (Sim)-binding sites. In contrast to many ventral midline enhancers, the primary enhancer can direct lacZ expression in the ventral midline as well as in the neurogenic ectoderm without a canonical Simbinding site. Intriguingly, the impaired transcriptional synergy between Dl and either Zld or Bcd led to aberrant and abolished lacZ expression in the neurogenic ectoderm and in the ventral midline, respectively. These findings suggest that the two enhancer activities of the sog primary enhancer are functionally consolidated and geographically inseparable.

• Journal of Embryo Transfer
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• v.7 no.1
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• pp.41-47
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• 1992

1. In vitro system, LR and FSR accelerated and facilitated meiotic progression, and LH selectively improved cytoplasmic maturation which is required to promote the formation of a male pronucleus. 2. Caffeine (2mM) in the fetilization medium was required not only for inducing zona penetrating ability of boar also for developing to the male pronucleus of the penetrat- ing spermatozoa in vitro. 3. The germinal vesicle (GV)stage was observed for the first 17.6 hr;germinal vesicle break-down (GVBD)stage between 17.6~26.4 hr ;metaphase I (M-I)from 26.4 - 30. 9hr;anaphase I(A-I)ranged from 30. 9~33.4hr;telophase I(T-I) at 33.4~34.4hr; and metaphase II(M-II) at 34.4-48hr. 4. The addition of 10%(v /v) pig follicular fluid (pFF) to maturation media significantly increased the rate of nuclear maturation of pig oocytes (p<0.01), whereas the rate of nuclear maturation of pig oocytes among three different media did not differ. 5. The presence of a primary culture of POEC promotes in vitro development of early cleavage stage pig embryos.

• BMB Reports
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• v.48 no.10
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• pp.589-594
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• 2015

The shadow enhancer of the short gastrulation (sog) gene directs its sequential expression in the neurogenic ectoderm and the ventral midline of the developing Drosophila embryo. Here, we characterize three unusual features of the shadow enhancer midline activity. First, the minimal regions for the two different enhancer activities exhibit high overlap within the shadow enhancer, meaning that one developmental enhancer possesses dual enhancer activities. Second, the midline enhancer activity relies on five Single-minded (Sim)-binding sites, two of which have not been found in any Sim target enhancers. Finally, two linked Dorsal (Dl)- and Zelda (Zld)-binding sites, critical for the neurogenic ectoderm enhancer activity, are also required for the midline enhancer activity. These results suggest that early activation by Dl and Zld may facilitate late activation via the noncanonical sites occupied by Sim. We discuss a model for Zld as a pioneer factor and speculate its role in midline enhancer activity.

• Applied Biological Chemistry
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• v.36 no.6
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• pp.553-556
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• 1993

Soybean nuclear extracts were prepared to examine the expression of SEF3 (soybean embryo factors 3), which binds to the upstream region of soybean ${\beta}-conglycinin$ ${\alpha}'$ subunit gene and is presumed to be a trans-acting factor for the expression of the gene. The relative levels of SEF3 binding activity in nuclear extracts of maturing soybean embryos were determined using the SE3 DNA probe containing two AACCCA hexanucleotides for gel mobility shift assay. The SEF3 activity increased in developing embryos from 16 to 32 days after pollination, whereas the mobility of the SE3-SE3-SEF3 complex decreased. The mobility of the complex was increased by the treatment of nuclear extracts with alkaline phosphatese, which could be inhibited by phosphate. Formation of the SE3-SEF3 complex was not affected by the binding buffer pH between 6.8 and 8.5.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.39 no.3
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• pp.211-215
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• 1994

Lipoxygenase is involved in the formation of certain undesirable flavors of soybean products. Three isozymes(L-1, L-2 and L-3) of lipoxygenase have been identified in soybean seeds, and the three types of mutants lacking L-1, L-2 and L-3, respectively, were detected in the 1980's. In this paper, lipoxygenase activity was measured to investigate the response of lipoxygenase in organs and tissues during soybean development. There was no tendency according to genotypes between lipoxygenase lacking mutants and normal soybeans in lipoxygenase activity of leaf at $V_3$ and $V_5$ stage. Likewise, pod wall lipoxygenase was no difference among genotypes tested at R$_{6}$ stage. Seed coat lipoxygenase activity was similar among the lipoxygenase lacking mutants, while normal soybean was lower as compared with that of the lipoxygenase lacking mutants. Embryo and cotyledon lipoxygenase activity in the lipoxygenase lacking mutants was much lower than that of normal soybean, also there was large difference among lipoxygenase lacking mutants. Thus, the lipoxygenase null mutant showed very weak value although the lacking L-3 mutant had a large effect on developing embryo lipoxygenase activity. It was suggested that soybean lipoxygenase isozymes expressed in embryo may be different from those expressed in the pod wall and leaf tissues.

• The Korean Journal of Malacology
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• v.21 no.1
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• pp.25-31
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• 2005

The embryos of marine bivalves have been commonly used in bioassays for the quality assessment of marine environments. Although several standard protocols for developmental bioassay with bivalves have been already proposed, there have been few trials for applying these protocols in environmental assessment, or for developing new protocol with Korean species. So, there is a strong need to establish the standard bioassay protocols using bivalves commonly found in Korean waters. Prior to developing a new protocol, it is essential to know the optimum conditions for the reliable bioassay procedures. Here, we established the purpose of this study to determine the optimum bioassay conditions for successful development of a common mussel, Mytilus galloprovincialis. The conditions considered as critical for developmental bioassay, and determined in this study were; (1) temperature, (2) salinity, and (3) initial density of embryo. The optimal temperature for developmental bioassay of M. galloprovincialis was determined as $15^{\circ}C$. At this temperature, the required time for the embryo to become veliger larva was 48 hr. The acceptable range of salinity for the embryotoxicity test using M. galloprivincialis was from 30 to 35 psu, which was narrower than that of the natural habitat of adult populations. The optimum density of embryo at the beginning of bioassay was 100 embryos/ml. Over this density, the proportion of normally developed larvae decreased significantly. The results obtained in this study will serve as a basis for preparation of the standard bioassay protocol using embryo of M. galloprovincialis.