• Journal of Fisheries and Marine Sciences Education
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• v.28 no.5
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• pp.1231-1243
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• 2016

The ultrastructural study on oocyt development and the process of vitellogensis in the oocytes during oogenesis in female Pampus echinogaster were investigated by electron microscope observations. In the previtellogenic phase, in particular, several intermitochondrial cements appear in the cytoplasms of the chromatin nucleleolus oocyte and perinuclear oocyte. The number of intermitochondrial cements are associated with the multiplication of the number of mitochondria in the early developmental stage. In the early vitellogenic phase, the Golgi complex in the cytoplasm of the yolk vesicle oocyte is involved in the formation of yolk vesicles containing carbohydrate yolks. At this time, many pinocytotic vesicles containing yolk precursors (exogenous substances) by pinocytosis are observed in the cytoplasm near the region of initial formation of the zona pellucida. In the late vitellogenic phase, two morphological different bodies, which formed by the modified mitochondria, appeared remarkably in the yolked oocytes. The one is the multivesicular bodies and another is yolk precursors. The multivesicular bodies were transformed into the primary yolk globules, while yolk precursors were connected with exogeneous pinocytotic vesicles near the zona pellucida. After the pinocytotic vesicles were taken into yolk precursors, the yolk precursors were transformed into the primary yolk globules. Thereafter, primary yolk globules mixed with each other, eventually, they developed into secondary and tertiary yolk globules. In this study, vitellogenesis of this species occurred by way of endogenous autosynthesis and exogenous heteogenesis. Vitellogenesis occurred through the processes of endogeneous autosynthesis, involving the combined activity of the Golgi complex, mitochondria and multivesicular bodies formed by modified mitochondria. However, the process of heterosynthesis involved pinocytotic incorporation of extraovarian precursors (such as vitellogenin in the liver) into the zona pellucida (by way of granulosa cells and thecal cells) of vitellogenic oocytes.

• Journal of Embryo Transfer
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• v.32 no.3
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• pp.111-122
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• 2017

The objective of this experiment was to explore the effects of Roscovitine (Rosco) prior to in vitro maturation (IVM) of immature pig oocyte. Brilliant cresyl blue test has been used to select the good quality of oocyte. Specifically, the effects of Rosco exposure on nuclear and cytoplasmic maturation, diameter, intracellular glutathione (GSH) and reactive oxygen species (ROS), and embryonic development after parthenogenetic activation (PA) and somatic cell nuclear transfer (SCNT), and gene expression levels in SCNT embryos have been measured. Cumulus oocyte complexes (COCs) have been exposed in $75{\mu}M$ of Rosco for 22 and 44 h. The COCs that were matured in the IVM for 44 h without Rosco used as control group. Diameter of matured porcine oocytes 44 h culture with Rosco was significantly lower than 22 h culture with Rosco and control groups. GSH was higher in control group than 22 h and 44 h with Rosco but reduction of ROS in 22 h than 44 h with Rosco. In PA, exposure with Rosco 44 h oocytes group has been significantly lower than 22 h and control group in rates of maturation, cleavage and blastocyst formation. Similarly, in SCNT embryos rates of maturation, cleavage and formation of blastocyst have been also significantly lower in 44 h Rosco treated group than other two groups. SCNT embryos treated with Rosco 22 h showed greater expression levels of POU5F1, DPPA2 and NDP52Il mRNA compared with other two groups. Our results demonstrate that Rosco treatment with 22 h prior to IVM improves the development competence of porcine oocyte.

• Clinical and Experimental Reproductive Medicine
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• v.44 no.3
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• pp.126-131
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• 2017

Objective: Oocyte degeneration often occurs after intracytoplasmic sperm injection (ICSI), and the risk factor is low-quality oocytes. The follicular fluid (FF) provides a crucial microenvironment for oocyte development. We investigated the relationships between the FF volume aspirated from individual follicles and oocyte retrieval, oocyte maturity, oolemma stretchability, fertilization, and development. Methods: This retrospective study included data obtained from 229 ICSI cycles. Ovarian stimulation was performed according to a gonadotropin-releasing hormone antagonist protocol. Each follicle was individually aspirated and divided into six groups according to FF volume ( < 1.0, 1.0 to < 2.0, 2.0 to < 3.0, 3.0 to < 4.0, 4.0 to < 5.0, and ${\geq}5.0mL$). Oolemma stretchability during ICSI was evaluated using a mechanical stimulus for oolemma penetration, that is, the stretchability was assessed by oolemma penetration with aspiration (high stretchability) or without aspiration (low stretchability). Results: Oocyte retrieval rates were significantly lower in the < 1.0 mL group than in the ${\geq}1.0mL$ groups (46.0% [86/187] vs. 67.5%-74.3% [172/255 to 124/167], respectively; p< 0.01). Low oolemma stretchability was significantly more common in the < 1.0 mL group than in the ${\geq}1.0mL$ groups during ICSI (22.0% [13/59] vs. 5.8%-9.4% [6/104 to 13/139], respectively; p= 0.018). There was a relationship between FF volume and oolemma stretchability. However, there were no significant differences in the rates of fertilization, cleavage, ${\geq}7$ cells at day 3, and blastocyst development among all groups. Conclusion: FF volume is potentially associated with the stretchability of metaphase II oolemma during ICSI. Regarding oolemma stretchability, ensuring a uniform follicular size during ovarian stimulation is crucial to obtain good-quality oocytes.

• Journal of Animal Science and Technology
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• v.63 no.2
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• pp.272-280
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• 2021

Cumulus-oocyte complexes (COCs), which contain immature oocytes, are matured in vitro for in vitro embryo production. Oocyte and cumulus cells are then separated using hyaluronidase. To date, there have only been a few reported cases of the toxic effects of hyaluronidase on porcine oocytes. The aim of this study was to compare the effects of bovine testis-derived hyaluronidase and recombinant human hyaluronidase on oocyte denudation and quality. Porcine COCs were matured for 44 h and denuded using different hyaluronidase concentrations and exposure times. Then, oocytes were activated by electrical parthenogenesis. In experiment 1, COCs were denuded using bovine-derived, ovine-derived (Hirax), and human recombinant (ALT-BC4) hyaluronidases for 10 and 20 min. In experiment 2, bovine-derived and human recombinant (ALT-BC4 and ICSI Cumulase®) hyaluronidases were used to denude the COCs for 2 and 20 min. In both experiments the oocytes were all completely denuded, and there was no degeneration. Rate of embryo development was significantly increased in group treated ALT-BC4 for 2 min and not significantly different in other treatment groups. In general it slightly decreased with longer exposure times. These results have confirmed that different sources of hyaluronidase do not have detrimental effects on the quality of porcine oocytes and suggest that the human recombinant hyaluronidase ALT-BC4 is suitable for oocyte denudation with an increased blastocyst rate.

• Journal of Animal Reproduction and Biotechnology
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• v.37 no.1
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• pp.34-41
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• 2022

In vivo oocytes grow and mature in ovarian follicles whereas oocytes are matured in vitro in plastic culture dishes with a hard surface. In vivo oocytes show a superior developmental ability to in vitro counterparts, indicating suboptimal environments of in vitro culture. This study aimed to evaluate the influence of an agarose matrix as a culture substrate during in vitro maturation (IVM) on the development of pig oocytes derived from small antral follicles (SAFs). Cumulus-oocyte complexes (COCs) retrieved from SAFs were grown in a plastic culture dish without an agarose matrix and then cultured for maturation in a plastic dish coated without (control) or with a 1% or 2% (w/v) agarose hydrogel. Then, the effect of the soft agarose matrix on oocyte maturation and embryonic development was assessed by analyzing intra-oocyte contents of glutathione (GSH) and reactive oxygen species (ROS), expression of VEGFA, HIF1A, and PFKP genes, and blastocyst formation after parthenogenesis. IVM of pig COCs on a 1% (w/v) agarose matrix showed a significantly higher blastocyst formation, intra-oocyte GSH contents, and transcript abundance of VEGFA. Moreover, a significantly lower intra-oocyte ROS content was detected in oocytes matured on the 1% and 2% (w/v) agarose matrices than in control. Our results demonstrated that IVM of SAFs-derived pig oocytes on a soft agarose matrix enhanced developmental ability by improving the cytoplasmic maturation of oocytes through redox balancing and regulation of gene expression.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2003.10a
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• pp.124-124
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• 2003

• Applied Microscopy
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• v.28 no.4
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• pp.447-463
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• 1998

This study was carried out to investigate the histomorphological changes and the electrophoretic patterns of egg components, obtained from 100 of 2-year-old female catfishes (Silurus asotus). Female Korean catfishes collected in the vicinity of Chollabukdo have synchronous ovaries which discontinously ovulate eggs during the breeding season (from late May until early July). The fishes were killed by severing the spinal cord just posterior to the head after immobilization with tricaine methanesulfonate (MS 222). Especially, the light microscopic and ultrastructural changes of ooplasm and follicular membranes of oocytes, were examined by means of light and transmission electron microscopy. The size of the nucleoli and number of the yolk granules increased as the oocyte groved. Yolk granules were deposited in the oocyte as fluid Due to tile presence of large early and late maturing oocytes, their ovaries were enlarged, transparent, granular and greenish in color. As the percentages of fish in late maturing oocyte (LMO) and ripe oocyte (RO) stage increased from March to April, mean gonadosomatic index (GSI) values (19.95%) increased. Zona radiata changed from squamous into cuboid in shape in the early maturing oocyte (EMO) stage. Processes, microvilli, from the zona radiata and from the oocyte grow, and make contact with each other in the pore canals of the zona radiata during vitellogenesis, but are withdrawn as the zona radials becomes more compact and devoid of pore canals during oocyte maturation. Seasonal changes in the microscopic appearance of the ovaries were well correlated with those in both gonadosomatic index and macroscopic appearance. The main cytological changes such as increase in size of cell, nucleus, nucleolus, and increase in number of nucleoli and mitochondria demonstrated with electron microscopy in the previtellogenic oocytes of Korean catfish, provided evidences for important synthetic processes in an early preparatory phase of oocyte development. The electrophoretic pattern of major band in mature stage was much thicker (24 k, 66 k, 90-110k dalton) than that in previtellogenic phase.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.39 no.6
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• pp.472-479
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• 2006

The sexual maturity and reproductive cycle of the common squid, Todarodes pacificus captured from the East Sea, Korea, between January 2004 and January 2006, were investigated by documenting changes in the gonadosomatic index (GSI), gonad development, and oocyte size frequency distribution. The GSI of females began to increase in July, reached a maximum in August, and then gradually decreased. The GSI of males increased from July to March. Using gonad histological observations, we identified four oocyte developmental stages. The changes in GSI and gonad tissue resulted in the classification of the annual reproductive cycle into the following four successive stages: immature (April to June), growing (June to July), mature (July to August), and ripe and spawning (August to March). According to the oocyte diameter size frequency distribution in the ovary, this species appeared to have asynchronous oocyte development and one spawning time.

• Journal of Embryo Transfer
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• v.31 no.1
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• pp.9-12
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• 2016

Growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15) are oocyte-specific growth factors that regulate many critical processes involved in early folliculogenesis and oocyte maturation. In this study, effects of GDF9 and BMP15 treatment during in vitro maturation of porcine oocytes upon development after parthenogenetic activation were investigated. Neither GDF, BMP15 alone nor in combination affects the number and viability of cumulus cells or the rates of oocyte maturation and blastocyst development. However, the treatment of GDF9 on porcine oocytes increased the number of trophectodermal (TE) cells of blastocysts derived from activated oocytes (P<0.05). The treatment of BMP15 increased the cell numbers of both inner cell mass (ICM) and TE cells (P<0.05). The treatment with the combination of GDF9 and BMP15 further increased the numbers of ICM and TE cells, compared with GDF9 or BMP15 treatment alone (P<0.05). In conclusion, the treatment of GDF9 or BMP15 (or both) enhanced the quality of blastocysts via the increased number of ICM and/or TE cells.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.46 no.6
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• pp.835-842
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• 2013

Flathead flounder Hippoglossoides dubius is a commercially important fish in the coastal waters of Gangwon Province, which is its spawning ground and breeding habitat. A total of 1,669 gonads were sampled monthly from February 2011 to May 2013 to investigate ecological characteristics, such as variations in maturation and spawning by gonad index, visual maturity stage, histological observations and oocyte diameter. Males were numerically dominant over females in the fishing grounds year round. The spawning season was from January to April, and the peak was from February to March. Oocyte number as a measure of fecundity was between 27,372 and 915,209 eggs with a length range of 26.0-48.7 cm TL, while the largest oocyte grew to 0.9-1.4mm in egg diameter during its spawning season. The relationship between fecundity and total length was $F=0.0016TL^{5.2539}$. The smallest mature lengths of the females and males were 28.4 and 22.6 cm respectively, and the 50% mature lengths of females and males were 32.9, 26.9 cm respectively.