• Asian-Australasian Journal of Animal Sciences
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• v.32 no.7
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• pp.956-965
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• 2019

Objective: The main goal of this study was to provide a morphological indicator that could be used to select high-quality oocytes of appropriate meiotic and developmental capabilities in pig. The higher quality of immature oocytes, the higher success rates of in vitro maturation (IVM) and in vitro fertilization (IVF). Thus, prior to the IVM culture, it is important to characterize oocytes morphologically and biochemically in order to assess their quality. Two of the largest indicators of oocyte quality are the presence of cumulus cells and status of chromatin. To investigate the effects of porcine oocyte chromatin configurations on the developmental capacity of blastocysts, we assessed oocyte chromatin status according to follicle size and measured the developmental potency of blastocysts. Methods: To sort by follicle size, we divided the oocytes into three groups (less than 1 mm, 1 to 3 mm, and more than 3 mm in diameter). To assess chromatin configuration, the oocytes were assessed for their stages (surrounded nucleolus [SN] germinal vesicle [GV], non-surrounded nucleolus [NSN] GV, GV breakdown, metaphase I [MI], pro-metaphase II [proMII], and metaphase II [MII]) at different maturation times (22, 44, and 66 h). To assess the development rate, oocytes of each follicle size were subjected to parthenogenetic activation for further development. Finally, GV oocytes were grouped by their chromatin configuration (SN, SN/NSN, and NSN) and their global transcriptional levels were measured. Results: SN GV oocytes were more suitable for IVF than NSN GV oocytes. Moreover, oocytes collected from the larger follicles had a greater distribution of SN GV oocytes and a higher developmental capacity during IVM, reaching MII more quickly and developing more often to blastocysts. Conclusion: Porcine oocytes with high-level meiotic and developmental capacity were identified by analyzing the relationship between follicle size and chromatin configuration. The porcine oocytes from large follicles had a significantly higher SN status in which the transcription level was low and could be better in the degree of meiotic progression and developmental capacity.

• Development and Reproduction
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• v.12 no.3
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• pp.283-288
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• 2008

The Pseudogobio esocinus were caught at Wyuleo-ri, Gyeombaek-myeon, Boseong-gun, Jeollanamdo from April to May 2003. The fishes were incubated in transparent aquarium located at the laboratory of Chonnam National University, and their embryonic and larval development were observed. The fertilized eggs were spherical, semitransparent, and adhesive, and were $1.98{\pm}0.19mm$ (n=50) in diameter. The embryo, including 31$\sim$32 myotomes, hatched through egg membrane at 164 hrs after fertilization. The newly-hatched larvae were $4.61{\pm}0.83mm$ (n=10) in total length (TL). At that moment, yolk was not absorbed, and mouth and anus were not open. Star and spot shaped melanophores were distributed on the lens, and dorsal, ventral, and caudal parts. At 42 days after hatching, larva was $16.22{\pm}0.65mm$ (n=10) in TL. Melanophores were scaterred at head, back, and side parts. Morphological features of the embryo were transferred to juvenile stage showing similar features with those of the adult fish.

• Applied Microscopy
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• v.41 no.1
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• pp.61-67
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• 2011

The purpose of this study is to investigate the hemp (Cannabis sativa cv. Chungsam) seed structure and ultrastructure of food reserves by scanning and transmission electron microscopy. We examined the seed coat and embryo consisting of a hypocotyl-radicle axis and two cotyledons. The seed coat consisted of exotesta and endotesta. The exotesta was a mechanical layer with lignified and elongated cells, while endotesta of the underlying layers of the exotesta was consisted of two separated cell layers. The collapsed outer layer of endotesta showed the unique reticulate structures. In cotyledon cells, protein and lipid bodies occupied most of cytoplasm. Protein bodies varied in diameter from 1.8 to $5.0{\mu}m$ and possessed a protein matrix containing electron-dense globoid crystals. Numerous lipid bodies ranged from 0.8 to $3.0{\mu}m$ in diameter were distributed around the protein bodies. During the early stages of breakdown, protein bodies rapidly changed their shape into the granular feature, however, lipid bodies were gradually degradated and fused each other. The degeneration process of protein bodies and lipid bodies of cotyledon cells might be correlated with the reports which hemp seeds rapidly lose their ability to germinate.

• Reproductive and Developmental Biology
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• v.36 no.4
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• pp.255-260
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• 2012

The purpose of this study was to assess follicular viability and competence through in vitro culture of preantral follicles isolated from vitrified mouse whole ovaries. Mouse preantral follicles were enzymatically isolated from vitrified- warmed and fresh ovaries and cultured for 10 days followed by in vitro oocyte maturation. In vitro matured oocytes were fertilized and cultured to the blastocyst stage. Five minutes pre-exposure to vitrification solution of whole ovaries had significantly higher (p<0.05) oocyte survival and maturation rates than between 10 min exposure groups. Oocyte diameter was significantly smaller (p<0.05) in the 5 and 10 min exposure groups ($69.4{\pm}2.8$ and $67.8{\pm}3.1$) when compared to that of control group ($71.7{\pm}2.1$). There was no statistical significant difference in blastocyst development rates between vitrification group (8.6%) and the fresh control group (12.0%). The mean number of cells per blastocyst was significantly lower (p<0.05) in the vitrification group ($41.9{\pm}20.2$) than in the fresh control group ($55.1{\pm}22.5$). The results show that mouse oocytes within preantral follicles isolated from the vitrified whole ovaries can achieve full maturation, normal fertilization and embryo development.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.1
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• pp.36-40
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• 2007

Proteins and lipids not only provide a source of energy to the cell, but also play vital roles in modifying the physical properties and function of the biological membranes. In the present study, we investigated the biochemical constituents, viz. proteins and lipids, in growing oocytes of goat antral follicles during summer and winter seasons. Goat genitalia in phosphate buffered saline (pH 7.4) were brought to the laboratory within one hour of slaughter under aseptic conditions at $37^{\circ}C$. Oocytes were aspirated from normal small (<3 mm in diameter) and large (>3 mm) follicles and pooled for biochemical estimations. A significant increase in the amount of protein and lipid was observed with the growth of the oocyte. The amount of protein varied non-significantly with the season, while the amount of lipid varied significantly. The amounts of phospholipid, cholesterol, free fatty acid, and triglyceride increased with the growth of the oocyte, but no significant effect of season in these constituents was observed. Lysolecithin, sphingomyelin, and sterols were the polar lipids identified in both oocytes prepared from small follicles (small oocytes) as well as large follicles (large oocytes). In addition, the small oocytes also contained phosphatidyl serine, while large oocytes contained phosphatidyl glycerol phosphate and phosphatidyl inositol. Among non-polar lipids, triglycerides and long chain alcohols appear only in small oocytes and not in large oocytes. Monoglycerides, 1,2-diglycerides, 1,3-diglycerides and o-dialkyl glycerol ethers, fatty acids, fatty acid methyl esters, and wax esters were identified in both small and large oocytes. Information on biochemical composition of growing oocytes is relevant to oocyte and embryo competence, culture and cryopreservation.

• Korean Journal of Ichthyology
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• v.21 no.4
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• pp.247-252
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• 2009

The egg development and early life history of the Korean endangered fish Koreocobitis naktongensis (Cobitidae) were investigated. Eggs from the females were obtained by injecting 10 IU/g of human chorionic gonadotropin and inseminated with sperm by the dry method in the laboratory. The fertilized eggs were 1.0~1.2 mm in diameter and had no oil globules. The embryo began to hatch about 38 hrs after fertilization under water temperature of $20{\pm}1^{\circ}C$. The newly-hatched larvae were 2.6~2.8 mm in total length, and their mouth and anus were not yet opened. Seven days after hatching, the postlarvae were 4.5~5.2 mm in total length, and their yolk sacs were completely absorbed. They entered the juvenile stage when all fin-rays were formed at 50 days after hatching, and their total lengths were 21~27 mm.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.50 no.3
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• pp.287-295
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• 2017

Egg development and morphological change of larvae of the Sakhalin sole Limanda sakhalinensis were studied by observing specimens obtained in a rearing experiment from fertilized eggs to the juvenile stage. The wild broodstock was collected in January 2010 and kept in a circular water tank (${\O}1.5{\times}1m$) at a temperature of $14.5{\pm}0.5^{\circ}C$. Fertilized eggs ranged from 0.72 to 0.82 mm ($0.77{\pm}0.07mm$, $mean{\pm}SD$) in diameter. The eggs were spherical, transparent and adhesive demersal. The egg yolk was divided from the oocyte 10 min after fertilization (AF), and an embryo was formed in 36 h AF. More than 50% of the eggs hatched within 133 h AF. The mouth and anus did not open until $3.5{\pm}0.25mm$ total length (TL). At 4, days after hatching (AH), the fish became larvae 3.7 to 4.2 mm ($4.0{\pm}0.36mm\;TL$), yolk absorption was completed and the mouth began to open. The left eye moved upward and the nostril moved to the right at 39 days AH. These post-larvae ranged from 8.0 to 9.9 mm TL ($8.9{\pm}1.33mm\;TL$). At 50 days AH, the fish became juveniles ($12.4{\pm}1.20mm\;TL$) There were 70-72 dorsal fin rays, 55-56 anal fin rays, 11 pectoral fin rays, and 6 ventral fin rays and the juveniles adopted a benthic life.

• Journal of Plant Biotechnology
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• v.4 no.1
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• pp.29-32
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• 2002

The ability to form embryiod from callus in satsuma mandarin is low and unstable. In this study, the conditions of embryogenic calli induced from nucellar tissue for promotion of plant regeneration in satsuma mandarin were investigated. The calli of I, II and III line were divided into two sizes of 0.5 mm and 1.0 mm in diameter and two weight gradients of percoll at 40% and 50% though the filter mesh. The frequency of embryo formation from $\phi$ 1.0mm-40% was slightly higher than callus that from others. Adventitious embryoids developed to a globular stage were transferred to regeneration medium. In 'Miyagawa Wase', the embryos from I and II line developed into a heart stage from most of $\phi$ 0.5 mm- 40% and $\phi$1.0 mm-40% calli, but it failed in 'Sugiyama Unshu'. In the cultivar of 'Miyagawa Wase', 63% of adventitious embryos transferred to the regeneration medium developed into the heart stage from the most $\phi$ 1.0 mm-40% calli of I line, but of 'Sugiyama Unshu' failed in some calli condition. The embryoids from two callus lines developed further to shoots and plantlets, while the embriods from III line abnormal failed to regenerate in the cultivar. From these results, it is suggested that the plant regeneration from embryogenic callus in satsuma mandarin could be affected by callus conditions.

• Clinical and Experimental Reproductive Medicine
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• v.40 no.2
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• pp.83-89
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• 2013

Objective: To investigate outcomes of stimulated IVF cycles in which GnRH antagonist was omitted on the ovulation triggering day. Methods: A total of 86 women who underwent controlled ovarian hyperstimulation with recombinant FSH and GnRH antagonist flexible multiple-dose protocols were recruited and prospectively randomized into the conventional group (group A) or cessation group (group B). The GnRH antagonist, 0.25 mg/day of cetrorelix, was started when the leading follicle reached 14 mm in diameter and was continuously administered until the hCG triggering day (group A, 43 cycles) or until the day before hCG administration (group B, 43 cycles). The maturity of oocytes, fertilization rate, embryo quality, and implantation and clinical pregnancy rates were evaluated. Results: The duration of ovarian stimulation, total dose of gonadotropins, serum estradiol levels on hCG administration day, and number of oocytes retrieved were not significantly different between the two groups. The total dose of GnRH antagonist was significantly lower in group B than group A ($2.5{\pm}0.9$ vs. $3.2{\pm}0.8$ ampoules, p<0.05). There was no premature luteinization in any of the subjects. The proportion of mature oocytes and fertilization rate were not significantly different in group B than group A (70.7% vs. 66.7%; 71.1% vs. 66.4%, respectively). There were no significant differences in the implantation or clinical pregnancy rates. Conclusion: Our prospective randomized study suggested that cessation of GnRH antagonist on the hCG administration day during a flexible multiple-dose protocol could reduce the total dose of GnRH antagonist without compromising its effects on pregnancy rates.

• Clinical and Experimental Reproductive Medicine
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• v.13 no.1
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• pp.67-75
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• 1986

Follicle monitoring in the normal and clomiphene·stimulated cycles were analyzed in the Seoul IVF and ET (In vitro fertilization and embryo transfer) program. Ovarian follicular diameters were measured by the real·time sector scanner and plasma estradiol levels were assayed by radioimmunoassay methods during periovulatory period. The maximum follicular sizes of the clomiphene-stimulated and normal cycles were 21.1+-3.4mm and 19.2+-0.8mm, respectively. The peak levels of plasma estradiol in the clomiphene-stimulated and normal cycles were 10538+-553.6ng/ml and 298.3+-39.6pg/ml, respectively. Daily growth rate of the follicular diameters of the clomiphene-stimulated and normal cycles were 2.1mm and 1.9mm, respectively. Mean follicular number of the clomiphene-simulated and normal cycles were 2.28+-1.12 and 1.12+-0.21, respectively. There was a good statistical correlation between the mean follicular diameters and the plasma estradiol levels in the normal ovulatory and c1omiphene-stimulated ovulatory menstrual cycles (p<0.05). Our data revealad that the mean follicular diameter and the plasma estradiol level prior to HCG administration in IVF and program should reach at the level of 17.8+-3.0mm and 949.4+-487.1 pg/ml, respectively.