• Title/Summary/Keyword: OOCYTE

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Studies on the Fine Structures of Mouse Oocyte Whose Maturation has been suppressed in Vitro by Dibutyryl Cyclic AMP (Dibutyryl Cyclic AMP에 의해 成熟이 抑制된 Mouse 卵子의 微細構造에 관한 硏究)

  • 崔林淳
    • The Korean Journal of Zoology
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    • v.18 no.2
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    • pp.87-101
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    • 1975
  • Electron microscopic studies on the ultrastructure of the mouse oocyte were made to investigate the inhibition of germinal vesicle breakdown by dibutyryl cAMP. The nuclear membrane of the dibutyryl cAMP-treated oocyte is characterized by a decreased degree of folding, maintains the normal double membrane structure, and shows an increased occurrence of the nuclear pore. It is suggested that these may be related to the suppression of the maturation of oocytes at the germinal vesicle. Mitochondria in the control cell were shown to be spread evenly throughout the cytoplasm and structurally underdeveloped or transitionary having little cristae development. On the contrary, mitochondria in the treated oocyte were found to be localized mainly around the nucleus and to show a greater extent of cristae development. The oocyte treated with dibutyryl cAMP appears to have fewer and structurally simpler lysosomes as compared to the control. The Golgi complex in the control oocyte exhibits the typical granular and lamellar structure, whereas that in the treated cell is poorly developed. Many multivesicular bodies, tonofilaments, and free ribosomes were observed in the control as well as in treated cells. The microvilli become structurally irregular, and a development of the perivitelline space is apparent in the treated oocyte. It is concluded that there is no basic difference in the ultrastructure between the oocytes treated with dibutyryl cAMP for 24 hours in the medium and those collected directly from the follicle. However, the finding that dibutyryl cAMP induces a development of more pores along the nuclear membrane strongly suggests the possibility that this compound inhibits the maturation of oocytes by influencing the permeability of the nuclear membrane.

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Effect of Recipient's Age on the Pregnancy Outcomes in Oocyte Donation Program (난자공여 프로그램에서 난자수혜자의 연령이 임신율에 미치는 영향에 관한 연구)

  • Suh, C.S.;Oh, S.K.;Kim, S.H.;Choi, Y.M.;Kim, J.G.;Moon, S.Y.;Lee, J.Y.
    • Clinical and Experimental Reproductive Medicine
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    • v.24 no.2
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    • pp.167-177
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    • 1997
  • Oocyte donation program developed to reach the pregnancy in those patients suffering from premature ovarian failure or surgery induced menopause, particularly in their reproductive age. With technical advances and popularity of ART (assisted reproductive technology), the indication of oocyte donation program extended to low responders, and even to naturally menopaused patients that has led them quite successfully to getting in pregnancy. The purpose of this study was to evaluate which one is involved in the decline of fertility between the oocyte and uterine factor. One hundred five cycles of oocyte donation program were performed in 84 patients from Jan., 1993 to Dec., 1996. Oocytes were donated from healthy, young, fertile anonymous donors or relatives or infertile patients with supernumerary oocytes. The study population was divided into 3 groups according to the age of recipients. Group 1 was less than 35 years old, Group 2 was between 35 to 39 years old, and Group 3 was more than 39 years old. The results were as follows: The mean age of oocyte donor was $31.5{\pm}3.3$ (range; 25-36). The mean concentration of basal serum FSH and peak serum estradiol were not different among groups. The mean number of oocytes retrieved from donors, embryos transferred to recipients, and fertilization rate were not different among groups. The clinical pregnancy rate was 37.3% in Group 1, 31.6% in Group 2, and 31.6% in Group 3, respectively. The spontaneous abortion rate was 16.0% in Group 1, 16.7% in Group 2, and 16.7 in Group 3, respectively. The multiple pregnancy rate was 20.0% in Group 1, 16.7% in Group 2, 16,7% in Group 3, respectively, The implantation rate was 11.3% in Group 1, 10.3% in Group 2 and 10.0% in Group 3, respectively. All of the pregnancy outcomes were not different statistically among groups. In conclusion, endometrial receptivity does not seem to be impaired as age increases with transfer of good quality embryos and adequate endometrial preparation.

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The Signal Transduciton of Ginsenosides, Active Ingredients of Panax ginseng, in Xenopus oocyte: A Model System for Ginseng Study

  • Nah Seung-Yeol;Lee Sang-Mok
    • Proceedings of the Ginseng society Conference
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    • 2002.10a
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    • pp.66-83
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    • 2002
  • Recently, we have provided evidence that ginsenosides, the active components of Panax ginseng, utilize pertussis toxin (PTX)-insensitive $G{\alpha}_{q/11}-phospholipase\;C-{\beta}3(PLC-{\beta}3)$ signal transduction pathway for the enhancement of $Ca^{2+}-activated\;Cl^{-}$ current in the Xenopus oocyte (British J. Pharmacol. 132, 641-647, 2001; JBC 276, 48797-48802, 2001). Other investigators have shown that stimulation of receptors linked to $G{\alpha}-PLC$ pathway inhibits the activity of G proteincoupled inwardly rectifying $K^+$ (GIRK) channel. In the present study, we sought to determine whether ginsenosides influenced the activity of GIRK 1 and GIRK 4 (GIRK 1/4) channels expressed in the Xenopus oocyte, and if so, the underlying signal transduction mechanism. In oocyte injected with GIRK 1/4 channel cRNAs, bath-applied ginsenosides inhibited high potassium (HK) solution-elicited GIRK current $(EC_{50}:4.9{\pm}4.3\;{\mu}g/ml).$ Pretreatment of the oocyte with PTX reduced the HK solution-elicited GIRK current by $49\%,$ but it did not alter the inhibitory ginsenoside effect on GIRK current. Prior intraoocyte injection of cRNA(s) coding $G{\alpha}_q,\;G{\alpha}_{11}\;or\;G{\alpha}_q/G{\alpha}_{11},\;but\;not\;G{\alpha}_{i2}\;or\;G{\alpha}_{oA}$ attenuated the inhibitory ginsenoside effect. Injection of cRNAs coding $G{\beta}_{1{\gamma}2}$ also attenuated the ginsenoside effect. Similarly, injection of the cRNAs coding regulators of G protein signaling 1, 2 and 4 (RGS1, RGS2 and RGS4), which interact with $G{\alpha}_i\;and/or\;G{\alpha}_{q/11}$ and stimulates the hydrolysis of GTP to GDP in active GTP-bound $G{\alpha}$ subunit, resulted in a significant reduction of ginsenoside effect on GIRK current. Preincubation of GIRK channel-expressing oocyte in PLC inhibitor (U73122) or protein kinase C (PKC) inhibitor (staurosporine or chelerythrine) blocked the inhibitory ginsenoside effect on GIRK current. On the other hand, intraoocyte injection of BAPTA, a free $Ca^{2+}$ chelator, had no significant effect on the ginsenoside action. Taken together, these results suggest that ginsenosides inhibit the activity of GIRK 1/4 channel expressed in the Xenopus oocyte through a PTX-insensitive and $G{\alpha}_{q/11}$-,PLC-and PKC-mediated signal transduction pathway.

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Improved Method for Heterologous Expression of Ion Channels in Xenopus Oocyte: a PCR Shortcut to Oocyte Expression

  • Han-Seop Kim;Changho Lee;Eunpyo Moon;Churl K. Min
    • Animal cells and systems
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    • v.3 no.2
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    • pp.181-185
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    • 1999
  • Xenopus oocyte is one of the widely used heterologous expression systems of ion channels for electrophysiological studies. Here we describe a new method in which cRNA produced by polymerase chain reaction (PCR) and in vitro transcription is injected to express ion channels in oocytes. This method enables us (1) to eliminate all or a part of the untranslated region of the cDNA and to replace it with a known sequence which helps increase the expression level in oocytes, and (2) to use the PCR product for in vitro transcription without subcloning. Using this method, the expression level of one of the neuronal nicotinic acetylcholine receptors (nAChRs) $\alpha$$_{6}$ subtype in oocytes was systematically increased by more than 100-fold, which was confirmed both by the $\alpha$-Bungarotoxin ($\alpha$,/TEX>Bgt) binding assay and the current measurement.t.

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A Case Report about Pregnancy and Delivery with Primary Amenorrhea by Oriental Medicine in Oocyte Donation (원발성 무월경 환자의 난자공여 시술시 한약치료를 병행한 임신 및 출산 1례)

  • Koh, Ji-Eun;Lyou, Myung-Sook
    • The Journal of Korean Obstetrics and Gynecology
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    • v.30 no.2
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    • pp.144-152
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    • 2017
  • Objectives: The aim of this case is to report the effects of oriental medicine on one patient with primary amenorrhea for endometrial preparation and implantation. Methods: A patient with primary amenorrhea had symptoms of sleep disorder, diarrhea, colpoxerosis. For preparing endometrium and implantation in oocyte donation after one previous failure, she was treated by twice a day herb medication for 75 days. And we observed the effects of treatments by improvement of symptoms and following up endometrial proliferation ultrasonography. After implantation, for maintaining pregnancy and live birth, she was also treated by twice a day herb medication for 45 days. Results: After treatments, Symptoms of sleep disorder, diarrhea, colpoxerosis were improved and the thickness of endometrium was prepared for implantation in oocyte donation. So she was pregnant and gave birth to a healthy baby 36 weeks later. Conclusions: This case shows that oriental medicine has its effective implementation for the implantational surroundings on patients with primary amenorrhea in oocyte donation programs.

Ultrastructural Studies on Oocyte Development and Vitellogenesis associated with Follicle Cells in Female Scapharca subcrenata (Pelecypoda: Arcidae) in Western Korea

  • Kim, Sung Han
    • Development and Reproduction
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    • v.20 no.3
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    • pp.227-235
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    • 2016
  • Ultrastructural studies on oocyte development and vitellogenesis in oocytes, and the functions of follicle cells during oogenesis and oocyte degeneration were investigated to clarifyb the reproductive mechanism on vitellogenesis of Scapharca subcrenata using electron microscope observations. In this study, vitellogenesis during oogenesis in the oocytes occured by way of autosynthesis and heterosynthesis. Of two processes of vitellogenesis during oogenesis, the process of endogenous autosynthesis involved the combined activity of the Golgi complex, mitochondria and rough endoplasmic reticulum. However, the process of exogenous heterosynthesis involved endocytotic incorporation of extraovarian precursors at the basal region of the oolema of the early vitellogenic oocytes before the formation of the vitelline coat. In this study, follicle cells, which attached to the previtellogenic and vitellogenic oocytes, were easily found. In particular, the follicle cells were involved in the development of previtellogenic oocytes by the supply of nutrients, and vitellogenesis in the early and late vitellogenic oocytes by endocytosis of yolk precursors. Based on observations of follicle cells attached to degenerating oocytes after spawning, follicles of this species are involved in lysosomal induction of oocyte degeneration for the resorption phagosomes (phagolysosomes) in the cytoplasm for nutrient storage, as seen in other bivalves. In this study, the functions of follicle cells can accumulate reserves of lipid granules and glycogen particles for vitellogenesis from degenerating oocytes after spawning.

Ultrastructural Studies on Oocyte Differentiation and Vitellogenesis in the Oocytes of Female Kareius bicoloratus in Western Korea

  • Jun, Je-Cheon;Gang, Hee Woong;Lee, Ki-Young
    • Development and Reproduction
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    • v.22 no.3
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    • pp.213-223
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    • 2018
  • Ultrastructural studies on oocyte differentiation and vitellogenesis in the oocytes of female Kareius bicoloratus were investigated by transmission electron microscopy. The Golgi complex in the cytoplasm is involved in the formation of yolk vesicles that contain yolk carbohydrates in the yolk vesicle of oocytes in the early vitellogenic phase. In this phase, many pinocytotic vesicles (PVs), which are formed by pinocytosis, contain yolk precursors (exogenous substances). These substances are associated with exogenous heterosynthetic vitellogenesis. In yolked oocytes in the late vitellogenic phase, two morphologically different bodies, which formed by modified mitochondria, appear in oocytes. One is a multivesicular body (synthesized by autosynthetic vitellogenesis), and the other is a yolk precursor (an exogenous substance formed by heterosynthetic vitellogenesis). The multivesicular bodies (MVB) are taken into the yolk precursors (YP) and are transformed into primary yolk globules. However, after the YP mix with exogenous PVs near the zona pellucida, they are transformed into primary yolk globules. Vitellogenesis of this species occurs via endogenous autosynthesis and exogenous heterogenesis. Vitellogenesis occurs through endogenous autosynthesis, which involves the combined activity of the Golgi complex, mitochondria and MVB formed by modified mitochondria. However, heterosynthesis involves pinocytotic incorporation of extraovarian precursors (such as vitellogenin in the liver) into the zona pellucida (via granulosa cells and thecal cells) of the yolked oocyte.

Identification of Protein Candidates in Porcine Oocytes during In Vitro Maturation

  • Lee, Jae-Dal;Cui, Xiang-Shun;Im, Gi-Sun;Seong, Hwan-Hoo;Kim, Nam-Hyung
    • Reproductive and Developmental Biology
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    • v.32 no.2
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    • pp.71-79
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    • 2008
  • Surface-enhanced laser desorption and ionization time-of-flight mass spectrometry (SELDI-TOF MS) is one of the recently developed proteomic technologies which is based on capturing proteins and peptides by chemically modified surfaces and highly sensitive for the analysis of complex biological samples. In the present study, to gain insights into oocyte maturation and early embryo development, SELDI-TOF-MS was used to find the protein candidates that are specifically or prominently expressed in porcine oocytes at the in vitro matured metaphase II (MIIl) and germinal vesicle (GV) stages. By selected CM10 chip, 16 candidates were found to be up-regulated in GV stage oocytes compared with in MII stage oocytes, their molecular weights were 8,180 (2 candidates), 10,226 (5 candidates), 15,767 (5 candidates) and 16,770 (4 candidates) Da respectively. And the expression of 29 candidates were higher in MII than in GV stage oocytes, their molecular weight were 10,832 (3 candidates), 17,743 (8 candidates), 20,122 (3 candidates), 22,131 (3 candidates), 24,857 (7 candidates) and 33,507 (5 candidates) Da, respectively. The expression of selected 13 candidates (0.2 and 1.0 % error tolerances) were analyzed using real time RT-PCR. The proteins that differentially regulated during oocyte in vitro maturation in the pigs may be potential biomarkers of oocyte maturation and quality.

Ultrastructural Studies on Oocyte Development and Vitellogenesis During Oogenesis in Female Boleophthalmus pectinirostris

  • Chung, Ee-Yung;Choi, Ki-Ho;Jun, Je-Cheon;Choi, Moon-Sul;Lee, Ki-Young
    • Animal cells and systems
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    • v.13 no.1
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    • pp.49-57
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    • 2009
  • For the study of the reproductive mechanism associated with the process of vitellogenesis, oocyte development and vitellogenesis during oogenesis in female Boleophthalmus pectinirostris were investigated by electron microscopic observations. The ovary consists of a pair of saccular structures with many ovarian lobules. In the early vitellogenic oocyte, the Golgi complex plays an important role leading to the formation of yolk vesicles containing carbohydrate yolks. At this time many pinocytotic vesicles containing yolk precursors are observed in the cytoplasm near the region of initial formation of the zona radiata. In the late vitellogenic oocytes, the multivesicular bodies, which are formed by modified mitochondria, are involved in the formation of the primary yolk granules. Precursors of yolk granules and multivesicular bodies develop to primary yolk globules with participation of pinocytotic vesicles. After primary yolk globules mix with each other, they develop into secondary and tertiary yolk globules. Based on these findings, vitellogenesis of B. pectinirostris occurs by way of the processes of endogenous autosynthesis and exogenous heterosynthesis. The process of autosynthesis involves the combined activity of the Golgi complex, mitochondria, and multivesicular bodies. However, the process of heterosynthesis involves pinocytotic incorporation of extraovarian precursors into the zona radiata of vitellogenic oocytes by way of the thecal cell layers and granulosa cells.

Sexual Maturity and Reproductive Cycle of the Common Squid Todarodes pacificus in the East Sea of Korea (한국 동해 해역에서 채집된 살오징어 (Todarodes pacificus)의 성성숙과 생식주기)

  • Baek, Hea-Ja;Kim, Jae-Won;Cho, Yun-Jung;Kim, Su-Am
    • 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.