• Asian-Australasian Journal of Animal Sciences
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• v.31 no.9
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• pp.1420-1430
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• 2018

Objective: Epigallocatechin-3-gallate (EGCG) is a major ingredient of catechin polyphenols and is considered one of the most promising bioactive compounds in green tea because of its strong antioxidant properties. However, the protective role of EGCG in bovine oocyte in vitro maturation (IVM) has not been investigated. Therefore, we aimed to study the effects of EGCG on IVM of bovine oocytes. Methods: Bovine oocytes were treated with different concentrations of EGCG (0, 25, 50, 100, and $200{\mu}M$), and the nuclear and cytoplasmic maturation, cumulus cell expansion, intracellular reactive oxygen species (ROS) levels, total antioxidant capacity, the early apoptosis and the developmental competence of in vitro fertilized embryos were measured. The mRNA abundances of antioxidant genes (nuclear factor erythriod-2 related factor 2 [NRF2], superoxide dismutase 1 [SOD1], catalase [CAT], and glutathione peroxidase 4 [GPX4]) in matured bovine oocytes were also quantified. Results: Nuclear maturation which is characterized by first polar body extrusion, and cytoplasmic maturation characterized by peripheral and cortical distribution of cortical granules and homogeneous mitochondrial distribution were significantly improved in the $50{\mu}M$ EGCG-treated group compared with the control group. Adding $50{\mu}M$ EGCG to the maturation medium significantly increased the cumulus cell expansion index and upregulated the mRNA levels of cumulus cell expansion-related genes (hyaluronan synthase 2, tumor necrosis factor alpha induced protein 6, pentraxin 3, and prostaglandin 2). Both the intracellular ROS level and the early apoptotic rate of matured oocytes were significantly decreased in the $50{\mu}M$ EGCG group, and the total antioxidant ability was markedly enhanced. Additionally, both the cleavage and blastocyst rates were significantly higher in the $50{\mu}M$ EGCG-treated oocytes after in vitro fertilization than in the control oocytes. The mRNA abundance of NRF2, SOD1, CAT, and GPX4 were significantly increased in the $50{\mu}M$ EGCG-treated oocytes. Conclusion: In conclusion, $50{\mu}M$ EGCG can improve the bovine oocyte maturation, and the protective role of EGCG may be correlated with its antioxidative property.

• Applied Microscopy
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• v.29 no.4
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• pp.417-426
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• 1999

The present investigation is designed to provide basic information on fine structure of the skin of the sea bass, Lateolabrax japonicks in relation to study of epidermal change with environmental and physiological change. The skin of the sea bass is divided into the epidermal layer and dermal layer. Epidermal layer consists of supporting cells and unicellular glands. The supporting cells were classified into the superficial cell, intermediated cell and basal cell. Gland cells were classified into the mucous secretory cell and club cell which is more frequently observed. Superficial cell of epidermal layer is squamous or cuboidal and contains well-developed rough endoplasmic reticulum and the surface is covered with numerous microridges. Superficial cells are connected to another cell with membrane interdigitations and desmosomes. Intermediated cell is ovoid and the electron density is higher than the other supporting cells. Basal cell is cuboidal and has a well-developed mitochondria and membrane interdigitation. The mucous secretory cell has a numerous membrane bounded secretory granules. The cytoplasm of club cell is divided into cortex and medullar. The medullar cytoplasm has a nucleus, intracellular organelles and central vacuole, and the cortical cytoplasm has a well-developed tonofilament. Club cells are connected to another cell with well -developed membrane interdigitations and desmosomes.

• Korean Journal of Food Science and Technology
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• v.27 no.6
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• pp.915-920
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• 1995

This study was performed to investigate the effect of aluminum compound on the aluminum contents and histological change in brain tissue of rats. Seventy five male Sprague-Dawley strains were divided into five groups consisting of the control, 250 ppm $AlCl_3$ group, 500 ppm $AlCl_3$ group, 250 ppm $Al_2(SO_4)_3$ group, 500 ppm $Al_2(SO_4)_3$ group and kept on the diet for 2 weeks. The weight gain was increased by administration of $AlCl_3$ but decreased by administration of $Al_2(SO_4)_3$ as compared to control group. The aluminum contents in brain tissue of each group; 250 ppm $AlCl_3$ group, 500 ppm $AlCl_3$ group, 250 ppm $Al_2(SO_4)_3$ group and 500 ppm $Al_2(SO_4)_3$ group were 64.63, 102.21, 132.64 and 180.41 ppm, respectively. Aluminum accumulation in brain tissue was higher with administration of $Al_2(SO_4)_3$ than with administration of $AlCl_3$. In $AlCl_3$ administration group, multiple small intracytoplasmic granules and microvacuole were seen in large pyramidal cells of cortex and granulovacuolar degeneration. In $Al_2(SO_4)_3$ administration group revealed pollagis pallor, cellular pyknosis, microcavitation resulted from edema in deeper cortical layers were observed. Blue-pigmentation which represents the accumulation of aluminum was noted In granulovacuolar degeneration site in $Al_2(SO_4)_3$ administration group.

• Molecules and Cells
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• v.40 no.8
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• pp.558-566
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• 2017

Regular monitoring on experimental animal management found the fluctuation of ART outcome, which showed a necessity to explore whether superovulation treatment is responsible for such unexpected outcome. This study was subsequently conducted to examine whether superovulation treatment can preserve ultrastructural integrity and developmental competence of oocytes following oocyte activation and embryo culture. A randomized study using mouse model was designed and in vitro development (experiment 1), ultrastructural morphology (experiment 2) and functional integrity of the oocytes (experiment 3) retrieved after PMSG/hCG injection (superovulation group) or not (natural ovulation; control group) were evaluated. In experiment 1, more oocytes were retrieved following superovulation than following natural ovulation, but natural ovulation yielded higher (p < 0.0563) maturation rate than superovulation. The capacity of mature oocytes to form pronucleus and to develop into blastocysts in vitro was similar. In experiment 2, a notable (p < 0.0186) increase in mitochondrial deformity, characterized by the formation of vacuolated mitochondria, was detected in the superovulation group. Multivesicular body formation was also increased, whereas early endosome formation was significantly decreased. No obvious changes in other microorganelles, however, were detected, which included the formation and distribution of mitochondria, cortical granules, microvilli, and smooth and rough endoplasmic reticulum. In experiment 3, significant decreases in mitochondrial activity, ATP production and dextran uptake were detected in the superovulation group. In conclusion, superovulation treatment may change both maturational status and functional and ultrastuctural integrity of oocytes. Superovulation effect on preimplantation development can be discussed.

• Korean Journal of Oriental Medicine
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• v.8 no.1
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• pp.93-104
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• 2002

The following is a list of morphologic and genetic characteristics of Pinellia tuber. 1. The original plant of Pinellia tuber is Pinellia ternata$(T_{HUNB})$$B_{REIT}$. With regards to its external morphology, it is smaller than other Araceae species and its spadix is longer than its leaves, which trifurcate. 2. As regards its internal morphology, its mucous cell is elliptical and the vessel is helical or annular-shaped. Granules exist in abundance and in various shapes. 3. Distribution and size of laticifers are the key criteria on which to differentiate between domestic and imported Pinellia tuber. Laticifers are mainly distributed in the epidermis in domestic Pinellia tuber and in the cortical parenchyma in imported Pinellia tuber. The size of laticifers is somewhere between 1,3 and $8{\mu}m$ in diameter in imported Pinellia tuber bigger than its domestic counterpart. 4. RAPD markers display a great similarity in bands between domestic and Chinese Pinellia tuber. However, RAPD primers 352, 358, 365, 368 and 374 are distinctive markers for domestic Pinellia tuber. In the meantime, North Korean Pinellia tuber, morphologically similar to domestic Pinellia tuber, is genertically distinctive from its domestic counterpart in primers 354, 358, 365, 368, 374 and 379, a finding that supports the postulation that North Korean Pinellia tuber is tuber of another Araceae species.

• Korean Journal of Agricultural Science
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• v.48 no.3
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• pp.607-615
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• 2021

Cytokines are protein mediators that possess the ability to assist cell-to-cell communication in immune system-related activities. In general, pathogen endotoxins activate the release of inflammatory mediators, and with time, there is an increase in the cytokine levels in the body. Interleukin (IL)-6 mediates the acute-phase inflammatory response, and elevated IL-6 levels have been reported in peritoneal fluids of women with pelvic inflammation and endometriosis, thereby associating it with oocyte quality and infertility. To overcome subfertility or infertility in humans and animals, the present study was done to examine the effect of recombinant IL-6 on porcine oocytes matured in vitro and subsequently to determine the fertilization rate and embryo development. Porcine oocytes were incubated with varying concentrations of IL-6 (0 - 2 ㎍·mL^-1) for 44 h followed by in vitro fertilization and culturing of the oocytes. The oocytes or embryos were fixed with 3.7% paraformaldehyde (PFA) and stained with fluorescence dyes, and the meiotic spindle, chromosome organization, fertilization status and embryo development were subsequently assessed under a fluorescence microscope. We observed induction of an abnormal meiotic spindle alignment in the oocytes incubated with IL-6 compared to the control oocytes incubated without IL-6. Moreover, significantly decreased fertilization rates and embryo development were observed for oocytes incubated with IL-6 (p < 0.05). Thus, an increased IL-6 level during oocyte maturation could be associated with fertilization failure due to an aberrant chromosomal alignment and a disruption of the cortical granules. Taken together, our results indicate that successful assisted reproduction can be achieved by controlling the levels of inflammatory cytokines.

• Journal of Veterinary Clinics
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• v.26 no.6
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• pp.586-590
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• 2009

Horsehair worms (Chordodes koreensis) develop as parasites in the bodies of grasshoppers, crickets, cockroaches, and some beetles. Chordodes koreensis is an accidental parasite of humans, livestock, or pets and poses no public health threat. The male of Chordodes koreensis in the later larval stage from canine vomitus was investigated by the scanning and transmission electron microscopy. In cross sections, the body wall is composed of four components namely epicuticle, cuticle, epidermis, and muscle layers. The parenchymal tissue fills the rest of the body and surrounds the visceral organs such as intestine, and ventral nerve cord but testes were not found. The epicuticle is a thin superficial layer whose surface shows rows of polygonal elevations called areoles. The cuticle has 17 layers of collagenous fibers spirally wound about the long axis of the worm. The section through the cuticle reveals the layers of large fibers cut obliquely lengthwise, alternating with layers of fibers sectioned obliquely crosswise. The layers of large fiber formed a double helix about longitudinal axis of the worm. The epidermis is a single layer. The muscles were interrupted by the nervous lamella in the only midventral portion. The medulla of muscle plate is composed of lightly stained cytoplasm, mitochondria, weakly developed endoplasmic reticulum, and glycogen granules. Between the medulla of a cell and the plasmalemma lies a broad cortical zone of myofilaments. The circular muscles are absent. The characteristic feature of the cytoplasm is that there was no content in peripheral mesenchyme, but was an abundance of large clear vacuoles which give the cytosome a foamy appearance. The nucleus of mesenchyme is not easily identified in our specimens.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.8
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• pp.1112-1121
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• 2019

Objective: Gram-negative bacteria lipopolysaccharide (LPS) has been reported to be associated with uterine impairment, embryonic resorption, ovarian dysfunction, and follicle retardation. Here, we aimed to investigate the toxic effects of LPS on the maturation ability and parthenogenetic developmental competence of bovine oocytes. Methods: First, we developed an in vitro model to study the response of bovine cumulusoocyte complexes (COCs) to LPS stress. After incubating germinal vesicle COCs in $10{\mu}g/mL$ of LPS, we analyzed the following three aspects: the expression levels of the LPS receptor toll-like receptor 4 (TLR4) in COCs, activities of intracellular signaling protein p38 mitogen-activated protein kinase (p38 MAPK) and nuclear factor-kappa B (NF-${\kappa}B$); and the concentrations of interleukin (IL)-$1{\beta}$, tumor necrosis factor (TNF)-${\alpha}$, and IL-6. Furthermore, we determined the effects of LPS on the maturation ability and parthenogenetic developmental competence of bovine oocytes. Results: The results revealed that LPS treatment significantly elevated TLR4 mRNA and protein expression levels in COCs. Exposure of COCs to LPS also resulted in a marked increase in activity of the intracellular signaling protein p-p38 MAPK and NF-${\kappa}B$. Furthermore, oocytes cultured in maturation medium containing LPS had significantly higher concentrations of the proinflammatory cytokines IL-$1{\beta}$, TNF-${\alpha}$, and IL-6. LPS exposure significantly decreased the first polar body extrusion rate. The cytoplasmic maturation, characterized by polar body extrusion and distribution of peripheral cortical granules, was significantly impaired in LPS-treated oocytes. Moreover, LPS exposure significantly increased intracellular reactive oxygen species levels and the relative mRNA abundance of the antioxidants thioredoxin (Trx), Trx2, and peroxiredoxin 1 in oocytes. Moreover, the early apoptotic rate and the release of cytochrome C were significantly increased in response to LPS. The cleavage, morula, and blastocyst formation rates were significantly lower in parthenogenetically activated oocytes exposed to LPS, while the incidence of apoptotic nuclei in blastocysts was significantly increased. Conclusion: Together, these results provide an underlying mechanism by which LPS impairs maturation potential in bovine oocytes.

• Applied Microscopy
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• v.7 no.1
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• pp.21-43
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• 1977

This experiment was undertaken in order to localize the labeled dbcAMP (dibutyryl cyclic AMP) in oocytes whose development has been suppressed by cold dbcAMP for 6 or 19 hours in vitro. Mouse oocytes were obtained from the ovaries of 3-4 week old A strain female mice, by puncturing the Graafian follicles in the modified Krebs-Ringer bicarbonate salt solution under the dissecting microscope. Those oocytes which have intact germinal vesicle were cultured in the basic culture medium supplemented with 0.4% bovine serum albumin (BSA). Cultivation of the oocytes was carried out in a microtube developed by Cho (1974). The cultures were then incubated in a humidified 5% $CO_2$ incubator maintained at $37^{\circ}C$ for 6 or 19 hours (Donahue, 1968). DbcAMP was added to culture medium for a final concentration of 100ug/ml, and $^3H-dbc$ AMP (specific activity 13 Ci/mM) for a final concentration of $40{\mu}Ci/ml$ was also added to the medium. For electron microscopic autoradiography, those oocytes recovered from the culture were washed with phosphate buffer (pH 7.4), and immediately prefixed in a 2.5% glutaraldehyde overnight and postfixed for 2 hours at $4 ^{\circ}C$ in 1% osmium tetroxide in phosphate buffer with pH 7.4 (Palade, 1952). After fixation, the materials were dehydrated in graded alcohol series and embedded in Epon 812 mixture based on the standard procedures (Luft, 1961). The thin sections $600-700{\AA}$ thick were mounted on the grids of 200 meshes. The grids containing sections were coated with a nuclear emulsion Kodak NTB-3 and stored in a cold dark box (at $4^{\circ}C$) for 3 weeks. After exposure, the samples were developed with Kodak D-19 and stained with uranyl acetate and lead citrate. Routine observation was made with Hitachi HU-11E electron microsocope. The results of the observation were as followings: 1. It was found that the labeled dbcAMP penetrated the egg plasma membrane and dispersed at random in the cytoplasm. 2. It was also observed that most of the labeled dbcAMP was attached to microfibrillar lattices portion of the oocyte cytoplasm. There fore, it is presumed that the receptor of the dbcAMP is localized in the microfibrillar lattices of the oocyte. 3. It also seems that some other cell organells such as mitochondria, Golgi complex, cortical granules are not directly related to the action of the dbcAMP. 4. The labeled dbcAMP was neither observed in the membrane nor in the nucleus. Therefore, it seems that there is no relationship between the concentration of dbcAMP and the nuclear membranous permeability. 5. There was no difference in number of dbcAMP particles when oocytes were cultured for 6 hours and 19 hours. 6. However, it was observed that, in same of the oocytes suppressed in germinal vesicle by dbcAMP for 19 hours, cell organells were moved and concentrated to a small portion of the cytoplasm, and that the morphology of the organells greatly changed to an abnormal. form. Therefore, it is supposed that those oocytes were in the process of degeneration. From the above results, it is expected that dbcAMP penetrated the egg membrane and was bound to the receptor which seems to be located in the microfibrillar lattiees portion, and that this dbcAMP-receptor complex inhibited some enzyme system of the oocytes which are essential for the germinal vesicle breakdown.