• Journal of Embryo Transfer
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• v.28 no.2
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• pp.127-132
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• 2013

The objective of this study was to examine the effect of in vitro maturation (IVM) medium, cytochalasin B (CB) treatment during intracytoplasmic sperm injection (ICSI), and electric activation on in vitro development ICSI-derived embryos in pigs. Immature pig oocytes were matured in vitro in medium 199 (M199) or porcine zygote medium (PZM)-3 that were supplemented with porcine follicular fluid, cysteine, pyruvate, EGF, insulin, and hormones for the first 22 h and then further cultured in hormone-free medium for an additional 21~22 h. ICSI embryos were produced by injecting single sperm directly into the cytoplasm of IVM oocytes. The oocytes matured in PZM-3 with 61.6 mM NaCl (low-NaCl PZM-3) tended to decrease (0.05

• Journal of Animal Reproduction and Biotechnology
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• v.39 no.1
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• pp.48-57
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• 2024

Background: Cadmium (Cd) is toxic heavy metal that accumulates in organisms after passing through their respiratory and digestive tracts. Although several studies have reported the toxic effects of Cd exposure on human health, its role in embryonic development during preimplantation stage remains unclear. We investigated the effects of Cd on porcine embryonic development and elucidated the mechanism. Methods: We cultured parthenogenetic embryos in media treated with 0, 20, 40, or 60 µM Cd for 6 days and evaluated the rates of cleavage and blastocyst formation. To investigate the mechanism of Cd toxicity, we examined intracellular reactive oxygen species (ROS) and glutathione (GSH) levels. Moreover, we examined mitochondrial content, membrane potential, and ROS. Results: Cleavage and blastocyst formation rates began to decrease significantly in the 40 µM Cd group compared with the control. During post-blastulation, development was significantly delayed in the Cd group. Cd exposure significantly decreased cell number and increased apoptosis rate compared with the control. Embryos exposed to Cd had significantly higher ROS and lower GSH levels, as well as lower expression of antioxidant enzymes, compared with the control. Moreover, embryos exposed to Cd exhibited a significant decrease in mitochondrial content, mitochondrial membrane potential, and expression of mitochondrial genes and an increase in mitochondrial ROS compared to the control. Conclusions: We demonstrated that Cd exposure impairs porcine embryonic development by inducing oxidative stress and mitochondrial dysfunction. Our findings provide insights into the toxicity of Cd exposure on mammalian embryonic development and highlight the importance of preventing Cd pollution.

• Molecules and Cells
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• v.42 no.8
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• pp.589-596
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• 2019

${\beta}Pix$ is a guanine nucleotide exchange factor for the Rho family small GTPases, Rac1 and Cdc42. It is known to regulate focal adhesion dynamics and cell migration. However, the in vivo role of ${\beta}Pix$ is currently not well understood. Here, we report the production and characterization of ${\beta}Pix$-KO mice. Loss of ${\beta}Pix$ results in embryonic lethality accompanied by abnormal developmental features, such as incomplete neural tube closure, impaired axial rotation, and failure of allantois-chorion fusion. We also generated ${\beta}Pix$-KO mouse embryonic fibroblasts (MEFs) to examine ${\beta}Pix$ function in mouse fibroblasts. ${\beta}Pix$-KO MEFs exhibit decreased Rac1 activity, and defects in cell spreading and platelet-derived growth factor (PDGF)-induced ruffle formation and chemotaxis. The average size of focal adhesions is increased in ${\beta}Pix$-KO MEFs. Interestingly, ${\beta}Pix$-KO MEFs showed increased motility in random migration and rapid wound healing with elevated levels of MLC2 phosphorylation. Taken together, our data demonstrate that ${\beta}Pix$ plays essential roles in early embryonic development, cell spreading, and cell migration in fibroblasts.

• Journal of Embryo Transfer
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• v.9 no.1
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• pp.127-137
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• 1994

This study was carried out to investigate the inhibiting or promoting effect of fetal bovine serum fractionated by the molecular weight and to examine the effect of reconstruction of serum fractions on the development of 1- and 2-cell mouse embryos fertilized in vitro (IVEE) . The serum was separated by ultrafiltration or gel filtration methods and added in m-KRB medium for culture of IVFE. The developemental ability(cavitation and hatching) of embryos following culture of day 4 and 6 was compared among fractions. Small molecular weight fraction( <3 kDa) significantly inhibited the development of 1-and 2-cell IVFE to the blastocyst stages, compared with other fractions. One-cell IVFE were more sensitively damaged than 2-cell embryos by that fraction and arrested mainly at 2~4 cell stages. Moreover, small amount(<3%,v /v) of the inhibiting fraction acted even with protein rich fraction(100~30 kDa) and arrested the embryonic development. On the other hand, 100~30 kDa fraction promoted the embryonic development and no inhibiting effect was observed at the level of 50%(v /v) in culture medium In the experiment of gel filtraton, =30 kDa fraction showed the highest promoting effect on the embryonic development, but <4 kDa fraction inhibited significantly the development. These results suggest that serum contains not only small molecular weight inhibitory component(s) but also promoting one rather than albumin on embryonic development. And serum can be more effectively used in the IVF program after removal of inhibitory component(s) by one of above separation methods.

• Journal of Animal Reproduction and Biotechnology
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• v.35 no.2
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• pp.163-170
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• 2020

Partheno Embryo's research is known to play a very important role in identifying the development of embryonic cells or analyzing the genetic mechanisms of embryonic development, but the information on apoptosis formed during the early stage of development on Partheno Embryo is very little. Therefore, this study analyzed whether the embryonic cell death of unit embryos can be inhibited by adding Scriptaid, one of HDACi, which plays a role in demethylation of histone proteins as a method of regulating the cell cycle in the early embryo development of Partheno Embryo. As a result, the differentiation rate was higher in the group that added Scriptaid and FBS, but the cellular development was higher in the group that added pregnant serum to Scriptaid. As a result of analyzing the expression of the gene through IF and PCR, the group with the addition of gestational serum increased the expression of BCL2 and PCNA, which affects the anti-Casp3 action in cell survival. In addition, it is interpreted that treatment of Scriptaid for 16 hours, rather than 24 h treatment lowers the expression of Casp-3, a representative factor of apoptosis, and also increases embryonic development, thus affecting early embryo development. Therefore, it is concluded that the 16-hour treatment of Scriptaid and the use of gestational serum will inhibit cell death in the early embryonic development and increase the development rate of the embryo.

• Journal of dental hygiene science
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• v.20 no.2
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• pp.74-81
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• 2020

Background: In the early stages of development, teeth and lingual papillae are induced and developed through special and complex epithelial-mesenchymal interactions. Tooth completion indicates the beginning of the weaning phase, and accordingly, many oral tissues and organs are completed, and it is thought that their developmental completion times are related to each other. The purpose of this study was to clarify the embryonic and neonatal development of the filiform papillae and mandibular molar tooth, and discuss the developmental relationship between these organs by comparing the developmental completion times. Methods: Embryos at embryonic day 15 (EM15), 17 (EM17), and 21 (EM21) and mice at neonatal day 1 (NE1), 5 (NE5), 10 (NE10), and 21 (NE21) were used for experimentation. Tissues dissected from embryos and mice were fixed, and processed for histological analysis. Sections from the tissues were stained with hematoxylin and eosin for observation under a light microscope. Results: Based on the histological analysis results, the developmental process of the lingual epithelium covering the dorsal surface of the tongue was classified into three stages: initiation, morphogenesis, and functional. The development of the filiform papillae begins at EM17; undergoes rapid morphological changes in epithelial cells at EM21, PN1 and PN5, and reaches the functional stage at PN10, which is the sucking phase. Tooth development begins at EM13 or 15 and is completed at NE21 through prenatal and postnatal development. Conclusion: The development of the filiform papillae was initiated late and completed quickly through embryonic and neonatal development in comparison with the mandibular molar tooth. The filiform papillae are considered to play an important role in sucking rather than mastication as it is completed in the sucking phase.

• Korean Journal of Animal Reproduction
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• v.22 no.1
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• pp.51-59
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• 1998

This study was undertaken to investigate the effects of ITS and EGF on embryonic development of in vitro matured and fertilized bovine oocytes in culture medium su, pp.ementing with or without calf serum. When fertilized oocytes were cultured in TCM-199 containing 0, 0.1, 0.5 and 1.0\ulcorner/ml ITS with 5% calf serum, the rates of development to blastocyst stage and the cell number of blastocysts were not significantly different among all treatments. And also monolayer of cumulus/granulosa cells prepared in containing calf serum and ITS were no beneficial effects of embryonic development. On the other hand, when EGF was su, pp.imented to TCM-199 containing calf serum or calf serum free, embryonic development rates(24.0 2.8% to 29.2 1.7% or 8% to 9%) and cell number of blastocysts(p<0.05) were significantly increased compared with EGF-free(22.1 2.1 or 1.0%, p<0.05). But when fertilized oocytes were cultured with cumulus/granurosa cells in TCM-199 containing EGF and calf serum, the rate of embryos development to the blastocyst stage and cell number of blastocysts were not significantly different compared with EGF-free and any concentrations. These results showed that ITS and EGF was not improved the development of bovine embryo in vitro matured and in vitro fertilized with calf serum and/or monolayer of cumulus/granulosa cells.

• ALGAE
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• v.36 no.2
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• pp.147-154
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• 2021

Kelp life history pathways alternate between macroscopic sporophytes that produce spores and microscopic gametophytes that produce gametes. Occasionally, an alternative pathway is seen. This study examined the circumstances by which the high latitude estuarine bull kelp, Nereocystis luetkeana, foregoes the "free-living microscopic stages by releasing embryonic sporophytes directly from sori. Sori were collected from adult N. luetkeana sporophytes from eight locations within Kachemak Bay, Alaska in 2018 and 2020 to examine spatial and temporal development of embryonic sporophytes on sori. Distinctions were made between sori collected from first-generation and overwintered adults to assess the influence of parental age on embryonic sporophyte release. Further distinctions were made between sori collected from attached and drifting individuals to assess the influence of the status of parental attachment to substrate on embryonic sporophyte release. Inspection of propagules released from sori after 48-h incubations indicated that embryonic sporophytes were occasionally released alongside viable spores. Though embryonic sporophytes were released from sori as early as spring, it was not evident that they were bound by seasonal or spatial limits. The percent of propagules that were embryonic sporophytes ranged from 0% to 100% but were not significantly different between first-generation and overwintered adults, nor were they different between attached and drifting individuals. Nevertheless, the characteristic of directly releasing embryonic sporophytes from adult sporophytes might have ecological advantages for N. luetkeana.

• Clinical and Experimental Reproductive Medicine
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• v.48 no.4
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• pp.362-367
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• 2021

Objective: The impact of early mechanical removal of cumulus cells on fertilization and embryonic development is not yet precisely known. This study aimed to investigate the effects of early and late cumulus cell removal on fertilization, polyspermy, embryonic development potential, blastocyst development, and clinical outcomes. Methods: A prospective study was conducted of patients who underwent in vitro fertilization between September 2019 and October 2020. Sibling oocytes were randomly allocated after insemination to early cumulus cell removal at 6 hours (group I) and late cumulus cell removal at 16-18 hours (group II). If total fertilization failure (TFF) was determined to have occurred at early cumulus cell removal, rescue intracytoplasmic sperm injection (ICSI) was performed. Fertilization, embryonic development, and pregnancy outcomes were compared. Results: A total of 912 oocytes were assigned to group I (458 oocytes) and group II (454 oocytes). Fertilization, polyspermy, embryo quality, and pregnancy outcomes were not significantly different between both groups. Rescue ICSI enabled fertilization of 79.2% of the TFF oocytes. Conclusion: Early cumulus cell removal at 6 hours had no significant difference in fertilization, polyspermy, embryo development, or obstetric and perinatal outcomes compared to late removal. Early cumulus cell removal combined with early rescue ICSI may have the potential to help couples with TFF.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.3
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• pp.346-352
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• 2008

To investigate goose feather follicle development and difference among the dorsal, ventral, and thoracal tracts during embryonic stage, the present study was conducted on 180 embryos at different ages obtained from the Jilin White goose, a Chinese indigenous breed. The study indicated that the epidermis and dermis of goose embryo formed between embryonic day 10 (E10) and 12 (E12). The thickness of the epidermis remained unchanged until hatching; while the thickness of the dermis increased throughout embryonic development. The primary feather follicles formed around E13-E14 and there were no new primary feather follicles forming after E18. The secondary feather follicles formed coincidently at E18. The density of primary and secondary feather follicles on the ventral and thoracal tracts were significantly higher than those on the dorsal tract (p<0.05). For primary and secondary follicles, the diameter of the feather bulbs and the depth of the feather follicles on the dorsal tract were much greater than those on the thoracal and ventral tracts (p<0.01), respectively; while the difference between the ventral and thoracal tracts was not significant (p>0.05). It is concluded that the Jilin White goose is of a single-follicle group structure, differing from mammals which are of multiple-follicle group structure.