• Clinical and Experimental Reproductive Medicine
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• v.42 no.4
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• pp.156-162
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• 2015

Objective: To investigate fertilization and embryo quality of dysmorphic mature oocytes with specific morphological abnormalities obtained from intracytoplasmic sperm injection (ICSI). Methods: The fertilization rate (FR) and embryo quality were compared among 58 dysmorphic and 42 normal form oocytes (control 1) obtained from 35 consecutive ICSI cycles, each of which yielded at least one dysmorphic mature oocyte, performed over a period of 5 years. The FR and embryo quality of 441 normal form oocytes from another 119 ICSI cycles that did not involve dysmorphic oocytes served as control 2. Dysmorphic oocytes were classified as having a dark cytoplasm, cytoplasmic granularity, cytoplasmic vacuoles, refractile bodies in the cytoplasm, smooth endoplasmic reticulum in the cytoplasm, an oval shape, an abnormal zona pellucida, a large perivitelline space, debris in the perivitelline space, or an abnormal polar body (PB). Results: The overall FR was significantly lower in dysmorphic oocytes than in normal form oocytes in both the control 1 and control 2 groups. However, embryo quality in the dysmorphic oocyte group and the normal form oocyte groups at day 3 was similar. The FR and embryo quality were similar in the oocyte groups with a single abnormality and multiple abnormalities. Specific abnormalities related with a higher percentage of top-quality embryos were dark cytoplasm (66.7%), abnormal PB (50%), and cytoplasmic vacuoles (25%). Conclusion: The fertilization potential of dysmorphic oocytes in our study was lower, but their subsequent embryonic development and embryo quality was relatively good. We were able to define several specific abnormalities related with good or poor embryo quality.

• Korean Journal of Animal Reproduction
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• v.21 no.4
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• pp.355-362
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• 1997

This study was undertaken in an effort to product embryos through in vitro maturation(IVM), in vitro fertilization(IVF) and in vitro culture(IVC) after cryopreservation of immature and mature porcine oocytes. The experiments were conducted to investigate IVM rate of oocytes frozen with 3 different cryoprotectants and to examine IVF and IVC of frozen-thawed oocytes. The CEI(cumulus cells expansion index) after IVM of frozen-thawed immature oocytes was higher in oocytes frozen with PG＋PEG(propylene glycol plus polyethylene glycol) than those frozen with single cryoprotectant and this index was almost 90% of unfrozen oocyte's index(2.39 vs. 2.66). The IVF rate of all frozen oocytes was very low(68% of unfrozen oocytes) and the IVF rate of frozen immature oocytes was slightly higher than that of frozen mature oocytes(39.0% vs. 34.4%), but polyspermic penetration was higher in frozen immature oocytes(21.9% vs. 19.1%). The cleavage rate after IVF of frozen-thawed oocytes was 9.3% for frozen mature oocytes and 11.3% for frozen immature oocytes and this rate was significantly lower(P<0.05) than that of control(60.7%). The development to 8-cell stage was greatly lower in frozen mature oocytes than in frozen immature oocytes. The results indicate that the use of PG plus PEG as cryoprotectant may be very effective for vitrification of porcine oocytes and the frozen-thawed immature porcine oocytes can be used fro in vitro embryo production based on IVM, IVF and IVC system.

• Journal of Embryo Transfer
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• v.12 no.3
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• pp.315-324
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• 1997

This study was undertaken in an effort to develop a cryopreservation system of immature and mature porcine oocytes. For this aim, the experiments were designed to examine the effect of cryoprotectants and equdibration time on the viability of frozen-thawed oocytes by using trypan blue(TB) and fluorescene diacetate(FDA) test. The viability of frozen immature oocytes evaluated by TB test was slightly higher than that of frozen mature oocytes. The viability(25.O%) after IVM of frozen-thawed immature oocytes greatly decreased that(42.9%) of oocytes just after thawing, but it was higher than frozen-thawed mature oocytes(15.8%). When immature oocytes were equilibrated for 10, 20 and 30 minutes before freezing the oocyte viability was 20.0, 31.3 and 42.9%, respectively. There was a tendency for long equilibration before oocyte freezing to be more effective for the immature oocytes and a short equilibration time for mature oocytes. Although there was no difference in viability index of frozen oocytes hetween the viability test methods, the index of TB test was slightly higher than that of FDA test. The viability(FDA test) of frozen-immature oocytes with 3 different crtoprotectants was 22.2% for propylene glycol(PG), 9.3% for polyehtylene glycol(PEG) and 65.6% for PG+PEG, in which PG+PEG was more protective against freezing effect.

• Development and Reproduction
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• v.24 no.3
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• pp.225-230
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• 2020

This study reports the presence of mature and bi-sexual phase gonads in red spotted grouper, Epinephelus akaara after less than a year of cultivation in a commercial indoor tank and a net cage. In December 2018, juveniles were placed in an indoor tank and cultured for five months. In June 2019, the fish were transferred to a net cage and cultured until September. The rearing temperatures ranged from 19.86℃-24.65℃ in the indoor tank and 21.86℃-27.65℃ in the net cage. During the net cage culture period, specimens were randomly selected for histological gonad examination. The highest gonadosomatic index (GSI) value was measured in July (3.38±2.53), and dramatically decreased in August (0.44±0.21) and September (0.42±0.30). In July, some mature fish showed signs of vitellogenic stage oocyte development (vitellogenic and oil droplet stage oocytes), but immature fish were in an early developmental stage containing peri-nucleolus stage (PNS) oocytes. Bi-sexual phase gonads containing spermatocytes and spermatids were observed in the lumen and several PNS oocytes. By August and September, most specimens showed early-stage ovary development. However, mature testis (in August) and bi-sexual phase gonads (in September) were also observed. These results provide evidence for early puberty and hermaphroditism in the red spotted grouper.

• Development and Reproduction
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• v.17 no.4
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• pp.421-426
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• 2013

In this study, oocyte steroidogenesis are investigated in relation to oocyte development in the starry flounder, Platichthys stellatus, a marine multiple spawner. Vitellogenic (0.52 and 0.55 mm oocyte diameter) and mature oocytes (0.63, 0.66 and 0.71 mm oocyte diameter) were incubated in vitro in the presence of $[^3H]17{\alpha}$-hydroxyprogesterone ($[^3H]17{\alpha}$-OHP) as a precursor. Steroid metabolites were extracted from the incubated media and oocytes, the extracts were separated and identified by thin-layer chromatography (TLC), high performance liquid chromatography (HPLC) and gas chromatographymass spectrometry (GC-MS). The major metabolites produced from $[^3H]17{\alpha}$-OHP were androgens [androstenedione ($A_4$) and testosterone (T)] and estrogens [$17{\beta}$-estradiol ($E_2$) and estrone ($E_1$)] and progestins [$17{\alpha},20{\alpha}$-dihydroxy-4-pregnen-3-one ($17{\alpha}20{\alpha}P$) and $17{\alpha},20{\beta}$-dihydroxy-4-pregnen-3-one ($17{\alpha}20{\beta}P$)] in vitellogenic and mature oocytes. The results from this study suggest the potential roles of $E_1$ in the oocytes with diameter 0.52-0.71 mm, $17{\alpha}20{\alpha}P$ and $17{\alpha}20{\beta}P$ at the oocytes of 0.63, 0.66 and 0.71 mm.

• Journal of Marine Life Science
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• v.7 no.1
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• pp.55-59
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• 2022

This study was conducted to provide basic information on the sexual mature and reproductive biology of Trichiurus lepturus. During the oogenesis, size of oocyte and nucleus showed the increase tendency but size ratio of nucleolus to nucleus was decreased. In the result of H-E stain, the stainability of the cytoplasm was changed from basophilic to eosinophilic. The egg diameter of initial vitellogenic oocyte was about 63.2 (±12.7) ㎛. In the cytoplasmic cortex, eosinophilic yolk nucleus was observed. The size of mature oocyte was 216.6 (±24.7) ㎛ and GVBD (germinal vesicle breakdown) was observed. The size of ripe oocyte was 317.9 (±80.9) ㎛ and the thickness of zona radiata was 4.2 (±1.7) ㎛. The oocyte developmental type is considered to belong to the group synchronous type. The accumulation of yolk in the oocyte, like most teleost, is thought to be of two types: exogenous and endogenous accumulation.

• Animal cells and systems
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• v.12 no.4
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• pp.313-319
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• 2008

The ultrastructure of oocytes during oogenesis and oocyte degeneration associated with follicle cells in female Sinonovacula constricta(Lamarck, 1818) were investigated by electron microscope observations. Ovarian follicles are surrounded by a matrix of vesicular connective tissue cells(VCT cells). VCT cells contain large quantities of glycogen particles and several lipid droplets in their cytoplasm. It is suggested that VCT cells act as a source of nutrients for vitellogenesis during oogenesis. In early vitellogenic oocytes, several coated vesicles, which appear at the basal region of the oocyte, lead to the formation of membrane-bound vesicles via endocytosis. The uptake of nutritive materials in coated vesicles formed by endocytosis appears through the formation of coated pits on the oolemma during vitellogenesis. During the late stage of oogenesis, yolk precursors(yolk granules), mitochondria and lipid droplets are present in the cytoplasm of late vitellogenic oocytes. In particular, proteinaceous yolk granules containing several different components are intermingles and form immature yolk granules. In the mature oocyte, small immature yolk granules are intermingled and form large mature yolk granules. Vitellogenesis occurs through a process of autosynthesis, involving combined activity of the Golgi complex, mitochondria and rough endoplasmic reticulum in the cytoplasm of vitellogenic oocytes. The process of heterosynthesis is where extraovarian precursors are incorporated into oocytes by endocytosis at the basal region of early vitellogenic oocytes before the formation of the vitelline coat. Follicle cells appear to play an important role in vitellogenesis and oocyte degeneration. The functions of attached follicle cells to the oocyte during oocyte degeneration are phagocytosis and digestion of phagosomes originating from oocyte degeneration. After digestion of phagosomes, it is assumed that the function of follicle cells can permit a transfer of yolk precursors necessary for vitellogenesis and allows for the accumulation of glycogen and lipid during oocyte degeneration, which can be employed by vitellogenic oocytes. Follicle cells of S. constricta may possess a lysosomal system for induction of oocyte breakdown and might resorb phagosomes in the cytoplasm for nutrient accumulation during oocyte degeneration.

• Applied Microscopy
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• v.18 no.1
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• pp.77-91
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• 1988

Since the Urechis unicinctus-oocyte grows asynchronously in the body fluid, various oocytes in developmental stages can be prepared from each individual. The oocytes obtained from the coelomic fluid are then classified into five developmental stages according to the fine structural features. The earlier oocytes (${\sim}18{\mu}m$) form cluster and thereafter the oocytes grow singly without a distinct support of somatic cell, such as accessory cell or matrix cell. The yolk granules begin to appear already in the oocyte of cluster stage, however, the typical yolk was observed at the stage IV. Therefore, it was suggested that the yolk deposition is correlated with the coelomic fluid. The mature oocyte measured about $150{\mu}m$ produces the invagination not only on oolemma(indentation) but also on nuclear envelope. After the formation of the indentation, the mature ooytes are stored in storge sacs. The fine structural features were combined in aspect of structural concept of light microscopical observation.

• Journal of fish pathology
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• v.16 no.3
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• pp.215-217
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• 2003

This study examined whether the connexin (Cx) is an essential protein during oocyte maturation in the ovary of the goldfish (Carassius auratus). In mature female goldfish ovaries, at late vitellogenic stage, human insulin-like growth factor-I (IGF-I; 20 M) and human chorionic gonadotropin(hCG; 20 IU/㎖) were injected. Twelve hr after the injection, mature female goldfish ovaries were removed and stored at -80C until analysis by RT-PCR. From the goldfish Cx43 cDNA sequence (GenBank accession number AB078505), two degenerate primers were designated. In vivo, 12 hr after the treatment with hCG, goldfish Cx43 mRNA expression level was increased, while the levels of IGF-I was not changed. Goldfish Cx43 mRNA expressed after, but not before the hCG treatment. These results suggest that Cx43 mRNA was judged to be a gene, which was transcribed during oocyte maturation induced by hCG.

• Journal of fish pathology
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• v.16 no.2
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• pp.125-129
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• 2003

Conncxin (Cx) is an important and essential protein induction of oocyte maturation, which is present in almost all mammalian tissucs except cirulating blood cells and adult skeletal museles. In this study, goldfish Cx43 cDNA sequence is available in GenBank under the accession number AB078505. Homology analyses using the GenBank and EMBL general database searches indicated that goldfish Cx43 cDNA has a high homology with carp Cx43 (95.1% identity), zebrafish Cx43 (90.5% identity), and chicken Cx43 (81.9% identity). Goldfish Cx43 is similar to the Cx family in its general features, and all the typical Cx consensus sequences are also found. Moreover, significantly increased Cx43 transcrpts were observed in mature goldfish (GSI; 18.3-21.7) pituitary and ovary when compared with immature goldfish (GSI; 4.9-6.0). Cx43 transeripts were weakly detectrd in both liver and kidney of immature and mature goldfish. There is possible that Cx43 nctivity was relation to oocyte maturation in the goldfish.