• Title/Summary/Keyword: oocyte diameter

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Assessment of In Vitro Oocyte Maturation in Two Gobiid Fish Species, Chasmichthys dolichognathus and Tridentiger trigonocephalus after Exposure to Benzo[a]pyrene

  • Hwang, In-Joon;Baek, Hea-Ja
    • Development and Reproduction
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    • v.15 no.3
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    • pp.223-230
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    • 2011
  • Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous environmental contaminants derived from incomplete combustion of carbons and crude oil. In this study, we investigated the effects of benzo[a]pyrene (B[a]P), a representative PAHs on in vitro sex steroid hormone production and germinal vesicle breakdown (GVBD) using isolated oocytes of longchin goby (Chasmichthys dolichognathus) and chameleon goby (Tridentiger trigonocephalus). Oocytes in diameters of 0.8-0.9 (end vitellogenic stage) and 0.9-1.0 mm (germinal vesicle migratory stage) from longchin goby and 0.5 mm (fully vitellogenic stage) from chameleon goby were used. In GVBD assay, B[a]P at 10 nM stimulated GVBD in the oocytes of 0.8-0.9 mm from longchin goby. B[a]P at 1 nM stimulated GVBD in the oocytes with diameter 0.5 mm from chameleon goby. In steroid production from oocytes of longchin goby, B[a]P at 100 nM decreased testosterone (T) production, B[a]P at 1,000 nM increased estraiol-17 (J (E2) production and 10 and 100 nM increased $17,20{\beta}$-dihydroxy-4-pregnen-3-one ($17{\alpha}20{\beta}P$) production in the oocytes with diameter 0.8-0.9 mm. B[a]P at 1,000 nM increased E2 production, 100 and 1,000 nM increased $17{\alpha}20{\beta}P$ production in the oocytes with diameter 0.9-1.0 mm. In steroid production of oocytes from chameleon goby, B[a]P at 1,000 nM increased $E_2$ production. B[a]P at 10 nM increased $17{\alpha}20{\beta}P$ production. In the ratio of $E_2$ to T ($E_2$/T), B[a]P at 100 and 1,000 nM increased $E_2$/T in the oocytes of longchin goby. B[a]P at 100 nM also increased $E_2$/T in the oocytes of chameleon goby. Taken together, these results suggest that B[a]P have not only weak estrogenic effects but progestogenic effects on oocyte maturation.

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.

Morphological Assessment of Ovulated and In Vitro Immature Canine Oocytes and Biological Availability according to the Size at Different Reproductive Stages

  • Hossein Mohammad-Shamim;Kim Min-Kyu;Jang Goo;Fibrianto Yuda-Heru;Oh Huyn-Joo;Kim Hye-Jin;Kim Joung-Joo;Kang Sung-Keun;Lee Byeong-Chun
    • Reproductive and Developmental Biology
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    • v.30 no.2
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    • pp.99-105
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    • 2006
  • The growing oocytes become progressively capable of resuming meiosis, and full meiotic competence appear when they are about 80% of the size of fully grown oocytes. As hormonal influences vary at different stages of reproductive cycle, the size of oocytes may vary according to the reproductive stages. The present study was designed to compare the diameter between the ovulated and freshly collected immature canine oocytes. The ovulated oocytes were collected 72 hr after ovulation by oviductal tube flushing by laparotomy under general anesthesia. Immature oocytes were collected by ovarian slicing method. Diameter of all oocytes was measured directly using epiflurescence microscope with a calibrated micro-eyepiece micrometer at ${\times}200$ magnification. The thickness of zona pellucida and diameter of cytoplasm were measured separately and recorded. A total of 2209 zona intact oocytes were collected, among them 628 from anestrus, 675 from follicular, 838 from luteal and 68 by fallopian tubes flushing methods. The average number of oocytes was 104.7, 168.8, 119.7 and 11.3 for anestrus, follicular, luteal and fallopian tubes flushing methods, respectively. The average diameters of the ooplasm and oocyte were significantly varied in different reproductive stages as well as with ovulated oocytes (P<0.05). The average diameter of ooplasm and oocyte was 115.6 and 127.7, 143.0 and 162.0, 134.6 and 150.6, 159.6 and 185.6 for anestrus, follicular, luteal and ovulated oocytes, respectively. Highest number of oocytes with larger diameter could be collected from the follicular and luteal stages. In conclusion, the follicular and luteal ovaries are the best sources of oocytes for canine IVM.

Ultrastructural Study on the Maturation of Oocyte in the African Giant Snail, Achatina fulica (아프리카 왕달팽이 (Achatina fulica) 난모세포 성숙에 관한 미세구조)

  • Chang, Nam-Sub;Han, Jong-Min
    • Applied Microscopy
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    • v.30 no.4
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    • pp.367-376
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    • 2000
  • The observation using an electron microscope shows that the maturation of the oocyte of African giant snail, Achatina fulica, proceeds over three stages. The oocyte of stage 1 is a small elliptic cell $(220\times400{\mu}m)$ whose light nucleoplasm contains two nucleoli. In its cytoplasm, a number of mitochondria, rough endoplasmic reticula, and ribosomes are found, while yolk granules are not. The nucleus of the oocyte of stage 2 is relatively large in comparison with the volume of cytoplasm, and contains one nucleolus. In the nuclear envelope comprising inner and outer double membrane, there are found a lot of nuclear pores for materials to pass through. A number of mitochondria, Golgi complex and lipid yolk granules appears in the cytoplasm, and proteinous yolk granules begin to form and mature in the vacuoles of various sizes ($0.8\sim3.0{\mu}m$ in diameter). The oocyte of stage 3 has an enlarged nucleolus. Material transportation through nuclear pore is not found any longer. The cytoplasm in this stage is filled with proteinous and lipid yolk granules. The microvilli are developed around the egg plasma membrane.

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Histological Description of Oogenesis in Largehead Hairtail Trichiurus lepturus (Teleostei: Trichiuridae) (갈치 Trichiurus lepturus 난자형성과정의 조직학적 기재)

  • Shin, So Ryung;Kim, Hyeon Jin;Oh, Han Young;Kim, Jae Won;Lee, Jung Sick
    • 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.

Porcine Oocyte Diameter in Relation to Maturational and Development Competence (돼지 난자의 직경이 체외성숙 및 체외발달에 미치는 영향)

  • ;;;;BN Day
    • Journal of Embryo Transfer
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    • v.14 no.1
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    • pp.17-22
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    • 1999
  • To investigate the maturational and development competece of porcine oocytes of different diameter groups, oocytes were obtained by aspiration from slaughterdhouse ovaries. After washing three times in NCSU23 medium, each cumulus-oocyte complex was transferred into a $8{mu}ell$ drop of the maturation medium (one oocyte per drop) under paraffin oil. The diameter without zona pellucida of oocytes was measured with micor-calibrator (Mikrometer, E. Leitz) on a screen connected to a VCR on an inverted microscope $(200\times)$. After being measured, the oocytes were divided into 6 groups according to their diameter size : <105, 105 to < 110, 110 to < 115, 115 to < 120, 120 to < 125 and > $125{\mu}{\textrm}{m}$, and in vitro maturation (IVM), fertillzation (IVF) and production (IVP) of oocytes / embryo was performed. The rates of in vitro maturation on oocytes in the greater 105 ${\mu}{\textrm}{m}$ size groups(91.8~100%) were significantly (P<0.05) higher than in the < 105 ${\mu}{\textrm}{m}$ group(66.7%). The rates of sperm penetration were significantly (P<0.05) low in < $105{\mu}{\textrm}{m}$ group (50.0%) than others groups (81.6~85.5%). But the plyspermic fertilization rate was significantly (P<0.05) higher in < $110{\mu}{\textrm}{m}$ oocytes groups than in the $110\leq{\mu}{\textrm}{m}$ size groups. The rates of cleavage and development to blastocysts rose as oocytes diameter increased, however, while oocytes over $120{\mu}{\textrm}{m}$ in diameter failed to develop to blastocysts. There results suggest that porcine oocytes have acquired full meiotic competece at a diameter of $105{\mu}{\textrm}{m}$ but not yet attended full development competence to blastocyst and that oocytes have acquired full development competence at a diameter of $110{\mu}{\textrm}{m}$.

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TLE-1 mRNA Expression during In Vivo and In Vitro Maturation in Porcine Oocytes (돼지 난자의 체내 및 체외 성숙시 Transducin-like Enhancer Protein 1(TLE-1) mRNA의 발현)

  • Jang, Ye-Jin;Kim, Dong-Woo;Lee, Yong-Seung;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Reproductive and Developmental Biology
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    • v.35 no.1
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    • pp.99-103
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    • 2011
  • Transducin-like enhancer protein 1(TLE-1) is protein associated with cell proliferation. This study analyzed change of TLE-1 mRNA expression during in vivo and in vitro maturation in porcine oocytes. Oocytes and granulose cells were collected from follicles of <2 mm, 2~6 mm and >6 mm in diameter in slaughtered pig's ovaries. Oocytes collected from follicles of 2~6 mm in diameter were used after in vitro maturation for 0, 10, 20 and 44 h. Cumulus cells and granulose cells were collected after treatment with hyaluronidase. In results, TLE-1 mRNA expression in oocytes collected from follicle >6 mm in diameter is increased, TLE-1 RNA expression in cumulus cells and granulosa cells from follicles <2 mm, 2 mm 6 mm and >6 mm in diameter. However, there is no significant difference. On the other hand, TLE-1 mRNA expression from oocytes cultured for 10 hand 44 h is increased, TLE-1 mRNA in cumulus cells cultured for 10 h is significant increased(p<0.05) than other culture periods. In conclusion, these results show that TLE-1 is expressed in all cell types of oocytes, cumulus cells and granulose cells, and associated with oocyte maturation.

Morphologic Parameters and in vitro Maturational Competence of Human Immature Oocyte Obtained from Stimulated IVF Cycle (미성숙난자의 형태학적 지표와 체외성숙능과의 관계)

  • An, So-Jung;Jee, Byung-Chul;Moon, Jeong-Hee;Lee, Jung-Ryeol;Suh, Chang-Suk;Kim, Seok-Hyun
    • Clinical and Experimental Reproductive Medicine
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    • v.35 no.4
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    • pp.285-291
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    • 2008
  • Objective: This study was performed to investigate whether cumulus morphology and oocyte diameter influence on in vitro maturation (IVM) of human germinal vesicle (GV) stage oocytes obtained from stimulated in vitro fertilization (IVF) cycles. Methods: Forty-one GV stage oocytes were obtained from 21 patients who received ovarian hyperstimulation and IVF. According to cumulus morphology before denudation, GV oocytes were classified into oocytes with dispersed cumulus cells (CCs) or compacted CCs. The diameters of denuded oocytes, both including and excluding the zona pellucida, were measured. All oocytes were cultured in commercial IVM medium. Maturation was defined as extrusion of the first polar body and the matured oocytes were inseminated by ICSI method. Results: Overall maturation and fertilization rate were 56.1% and 73.9%. Matured oocytes had significantly higher proportion of oocytes with dispersed CCs compared to oocytes failed to mature (91.3% vs. 55.6%, p=0.023). There were no significant differences of oocytes outer ($155.7{\mu}m$ vs. $152.4{\mu}m$, NS), inner ($114.3{\mu}m$ vs. $113.4{\mu}m$, NS) diameters and zona thicknesses ($41.3{\mu}m$ vs. $39.1{\mu}m$, NS) between matured and not-matured oocytes. In-vitro maturation rate of oocytes with dispersed CCs was significantly higher than which of oocytes with compacted CCs (67.7% vs. 20.0%, p=0.044). Oocyte diameters (outer and inner) and thicknesses were not related with maturational competence. Conclusion: Our results suggest that in vitro maturational competence of GV stage oocytes obtained from stimulated IVF cycles is closely associated with the cumulus morphology but not oocyte diameter.

Ultrastructural Study of Oogenesis and Reproductive Cycle of the Female Manila Clam, Ruditapes philippinarum in Komso Bay, Korea

  • Chung, Ee-Yung;Lee, C-Hang-Hoon;Park, Ki-Ho
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2001.02a
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    • pp.46-49
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    • 2001
  • R. Philippinarum is dioecious and oviparous. In the early vitellogenic oocyte, the Golgi apparatus and mitochondria present in the perinuclear region are involved in the formation of lipid droplets and in lipid granule formation. In the late vitellogenic oocyte, the endoplasmic reticulum, mitochondria in the cytoplasm are involved in the formation of proteid yolk granules. At this time, exogenous lipid granular substance and glycogen particles in the germinal epithelium are passed into the ooplasm of oocyte through the microvilli of the vitelline envelope. Ripe oocytes are about 55-60 $\mu$ m in diameter. The spawning period was once a year between early June and early October, and the main spawning occurred between July and August when seawater temperature was approximately 20 C. The reproductive cycle of this species can be categorized into five successive stages: early active stage (February to March), late active stage (April to May), ripe stage (April to August), partially spawned stage (June to October), and spent/inactive stage (August to March). Gonad developmental phases by histological qualitative analysis showed similar results with those of quantitative image analysis.

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Ultrastructural Description on Oogenesis of the Melania Snail, Semisulcospira libertina libertina (Gastropoda: Pleuroceridae) (다슬기, Semisulcospira libertina libertina의 난자형성과정에 관한 미세구조적 기재)

  • Kim, Eun-Kyoung;Lee, Jung-Sick
    • The Korean Journal of Malacology
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    • v.25 no.2
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    • pp.145-151
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    • 2009
  • The ultrastructural changes in germ cells during oogenesis of the melania snail, Semisulcospira libertina libertina have been investigated by light and electron microscopy. The ovary is located on the surface of the hepatopancreas in the spiral posterior region. The ovary exhibited greenish color in the gonadal mature season. The ovary was composed of a number of oogenic follicles. Oogenesis was divided into five stages with histological features: (1) oogonia, (2) previtellogenic, (3) initial vitellogenic, (4) active vitellogenic, and (5) mature stages. Oogonia were oval in shape, $4-6\;{\mu}m$ in diameter, and had a large nucleus. Previtellogenic oocytes were about $20\;{\mu}m$ in diameter and the cytoplasm reacted with hematoxylin in H-E satin. Initial vitellogenic stage, oocytes were $60-80\;{\mu}m$ in diameter, and small yolk granules of low electron density are scattered in the cytoplasm. Oocytes in the initial vitellogenic stage were connected with ovarian follicle by egg stalk. Active vitellogenic oocyte were $100-120\;{\mu}m$ in diameter. Electron density, size and quantity of yolk granules that are distributed in the cytoplasm have increased from the previous stage. Result of TEM observations, the oocyte contains well-developed Golgi complex, endoplasmic reticula and tubular mitochondria in the cytoplasm. Cytoplasm of mature oocyte was filled with proteinaceous yolk globules of high electron density. In this stage, the length of microvilli in the egg envelope was approximately $1.1\;{\mu}m$.

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