• Title/Summary/Keyword: oocyte diameter

검색결과 106건 처리시간 0.02초

성선 자극호르몬의 비율이 인간난자의 체외수정에 미치는 영향에 관한 연구 (The Significance of Gonadotropin Ratio in In Vitro Fertilization of Human Oocytes)

  • 문신용
    • Clinical and Experimental Reproductive Medicine
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    • 제15권2호
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    • pp.135-147
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    • 1988
  • To compare the stimulation effect of the ratio in follicle stimulating hormone and luteinizing hormone in induction of multiple follicular growth, the serum $E_2$ level, the diameter of follicle, number of aspirated follicles and cleavage rate of in vitro fertilized preovulatory oocytes as well as the pregnancy rate were evaluated. Forty one patients with irreparable tubal disease were stimulated by hMG(n=24) or FSH/hMG(n=17) for the purpose of in vitro fertilization and embryo transfer. The following results were obtained. 1. Serum estradiol($E_2$) levels on the day of hCG administration were $921.0{\pm}353.3\;pg/ml$ in hMG group and $1272.9{\pm}1060.6\;pg/ml$ in FSH/hMG group. The serum $E_2$ value of hMG group was significantly lower than that of FSH/hMG group. 2. The diameter of leading follicle by ultrasonogram on the day of hCG administration were $16.2{\pm}2.0\;mm$ in hMG group and $16.2{\pm}2.6\;mm$ in FSH/hMG group. No significant difference of follicle diameter between two groups was demonstrated. 3. The number of follicles with diameter above 10 mm by sonogram on the day of hCG injection were $3.91{\pm}2.32$ in hMG group and $6.52{\pm}3.86$ in FSH/hMG group. There was significant difference of number of follicles between two groups, (p< 0.01). 4. The number of oocytes found per patient at aspiration were $2.59{\pm}1.00$ in hMG group and 3. $76{\pm}2.31$ in FSH/hMG group. There was significant difference of number of aspirated oocytes between two groups. (p< 0.05). 5. The detection rate of preovulatory oocyte at aspiration were 68.4%(39/57) in hMG group (n=22) and 77.6%(38/49) in FSH/hMG group (n=13). 6. The cleavage rate of preovulatory oocyte at 44 hours after insemination were 74.4%(29/39) in hMG group(n=22) and 81.6%(31/38) in FSH/hMG group (n=13). When only hMG was used, one pregnancy was established in 15 patients to whom 29 zygotes were transferred. And a full term normal female baby was delivered by elective cesarean section. In the FSH/hMG group, five pregnancies out of 9 transferred patients were confirmed by serum ${\beta}-hCG$. Two pregnancies were spontaneously aborted before the 6th week of pregnancy. One patient aborted her baby at the 18th week of pregnancy because of incompetent internal os of the cervix. Two patients delivered two full term babies by elective cesarean section. From the above findings, paralell with the increase in the ratio of exogenous follicle stimulating hormone to luteinizing hormone, an increase in oocyte recovery was observed as well as an improvements in pregnancy rate. It was concluded that FSH enrichment early in the follicular phase had a beneficial effect in the controlled ovarian hyperstimulation.

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Seasonal Changes in Concentrations of Proteins and Lipids in Growing Goat Oocytes

  • Sangha, G.K.;Bhatia, H.;Khera, K.S.
    • Asian-Australasian Journal of Animal Sciences
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    • 제20권1호
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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.

기름가자미(Glyptocephalus stelleri) 성숙기 난모세포에서의 성스테로이드 호르몬 대사물질 분석 (Steroid Metabolism in the Blackfin Flounder Glyptocephalus stelleri during Oocyte Maturation)

  • 이해원;백혜자
    • 한국수산과학회지
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    • 제48권4호
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    • pp.483-488
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    • 2015
  • We studied oocyte steroidogenesis in the blackfin flounder Glyptocephalus stelleri as a region-specific species, in the East Sea of Korea during the spawning season. Maturing oocytes (0.76, 0.82, 0.88, and 0.91 mm in 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 medium and oocytes, and 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 (A4) and testosterone (T)] and estrogens [$17{\beta}$-estradiol (E2) and estrone (E1)] and progestins [$17{\alpha},20{\alpha}$-dihydroxy-4-pregen-3-one ($17{\alpha}20{\alpha}P$) and $17{\alpha}20{\beta}$-dihydroxy-4-pregnen-3-one ($17{\alpha}20{\beta}P$)] in maturing oocytes. The metabolic rate of $17{\alpha}20{\beta}$ was elevated (29.04%) in oocytes measuring 0.88 mm (nucleus migration stage following the induction of germinal vesicle breakdown), but was very low in oocytes measuring 0.76, 0.82, and 0.91 mm (0.42, 0.67, and 2.62%, respectively). From these results, we suggest that $17{\alpha}20{\beta}P$ acts as a maturation-inducing steroid in the blackfin flounder.

Surgical Oocyte Retrieval and the Developmental Potential of the Oocytes Derived from Prepubertal Calves

  • Yeh, S.P.;Ju, J.C.;Tseng, J.K.;Chou, P.C.;Chen, C.H.;Chih, T.;Fan, Y.K.
    • Asian-Australasian Journal of Animal Sciences
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    • 제17권2호
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    • pp.174-178
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    • 2004
  • The objectives of this study were to investigate the ovarian responsiveness of juvenile calves to exogenous gonadotropin treatments and to establish the oocyte retrieval technique for prepubertal heifers. Three 78-day-old calves were treated with 4 doses (40, 30, 30 and 30 mg) of FSH (Folltropin V) at 12 h interval up to 229 day-old. Surgical oocyte retrieval was performed 24 h after the last injection of FSH. Calves with good ovarian responses to FSH treatment had an average ovarian size of $5{\times}3$ cm compared to $3{\times}2$ cm in the less-responsive animals. Large variations were observed in the number of total follicles ($51{\pm}45$), aspirated follicles ($39{\pm}36$), oocytes recovered ($23{\pm}25$) and usable oocytes recovered ($11{\pm}19$) during 78 to 229 day-old. Oocytes derived from prepubertal calves had significantly lower maturation rate than those from cows (34 vs. 100%, p<0.05). Mean diameters of calf oocytes ($144{\pm}1{\mu}m$) and ooplasm ($110{\pm}1 {\mu}m$) were significantly lower than those of cows ($149{\pm}1$ and $25{\pm}1{\mu}m$, respectively). The diameter of the ooplasm also increased significantly after in vitro maturation (IVM) ($108{\pm}1$ vs. $112{\pm}1{\mu}m$). However, further studies are required to optimize the IVP system for the oocytes derived from prepubertal heifers.

Rat의 성주기에 따른 난포의 수와 형태변화 (Numerical and morphologic changes of ovarian follicles in each stage of estrus cycle in rats)

  • 이의주;곽수동
    • 대한수의학회지
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    • 제39권3호
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    • pp.455-462
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    • 1999
  • This study was designed to investigate the number of the growing and mature follicles in each stage of estrus cycle in mature rats. Eighteen mature rats(Sprague-Dawley, initially 190~230gm) were randomly alloted into 4 groups(proestrus, estrus, metestrus, and diestrus) according to estrus cycles. The uteri and ovaries of rats were collected and then alternative sections of paraffin embedding ovaries were stained with H-E. Numbers of large, middle and small follicles or only large and middle follicles from secondary and tertiary follicles were investigated by LM photography of preparations. Small follicles were defined as secondary follicles with 2~5 cell layers of granulosa cells surrounding the oocyte, and middle follicles were defined as secondary follicles with more than 5 cell layers or with early signs of antral cavity or with more than one small cleft on either side of the oocytes and large follicles were defined as tertiary follicles with a single medium or large antral cavity. The number of follicles in a pair ovary per rat was appeared to be ranged from 207 to 370 and the mean number of these follicles was $270.4{\pm}52.6$ and the mean number of follicles per ovary was $134.9{\pm}32.0$. The mean number of large, middle and small follicles per ovary was appeared to be $16.4{\pm}4.4$($12.2{\pm}3.3%$), $36.2{\pm}8.6$($26.8{\pm}6.4%$), and $82.7{\pm}24.0$($61.3{\pm}17.8%$), respectively. The mean number of large and middle follicles in each stage group of estrus cycle was appeared to be $17.8{\pm}2.1$ and $38.3{\pm}7.4$ at proestrus stage group, $15.7{\pm}5.2$ and $38.0{\pm}10.0$ at estrus stage group, $16.5{\pm}3.5$ and $33.8{\pm}7.0$ at metestrus stage group, $16.7{\pm}5.8$ and $29.7{\pm}5.5$ at diestrus group, respectively. In histological findings of large follicles during each estrus cycle, the large follicles in proestrus group contain single small antrum, thick granulosa cell layers, and were $300{\sim}500{\mu}m$ in diameter and were growing follicles with PCNA-positive cells in the granulosa cell layers, and other luteinizing follicles of proestrus cycle stage were decreased in size and were thicker in wall thickness and more luteinized than those in metestrus and diestrus stage groups. The large follicles in estrus stage group contain thick granulosa cell layers and nonprominent cumulus-oocyte complexes in antrum, and were $400{\sim}700{\mu}m$ in diameter and were growing follicles with PCNA-positive cells in the granulosa cell layers. The large follicles in metestrus and diestrus stage groups contain enlarged antrums, thinner layers of walls and prominent cumulus-oocyte complexes, and were $700-950{\mu}m$ in diameter, and were nongrowing follicles without PCNA-positive cells or another large follicles contain cells with dark stainability and distinct boundary.

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반복적인 과배란 처치 경험이 있는 한국흑염소에서 난포 자극 및 복강경을 이용한 난포란 채취(LOPU) (Follicular Stimulation and Laparoscopic Ovum Pick-up (LOPU) in Repeatedly Superovulated Korean Black Goats)

  • 이용범;이두수;조상철;신상태
    • 한국수정란이식학회지
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    • 제29권1호
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    • pp.35-41
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    • 2014
  • Laparoscopic ovum pick-up (LOPU) is a convenient method for collecting oocytes in small ruminants. LOPU has the advantage of being a less invasive means of oocyte collection, thereby allowing for a repeated usage of the oocyte donor animals. A total of 25 Korean black goats were used in the winter season (December to February) and LOPU was applied to the goats which had been treated for superovulation more than two times during the last twelve months. Estrus was synchronized with an intravaginal insert containing 0.3 g progesterone for 10 to 12 days. Ovaries were hyperstimulated with eCG 1,000 IU oneshot, FSH with eCG (50 mg / 1,000 IU; 70 mg / 500 IU; 70 mg / 1,000 IU) oneshot or FSH multiple-shot with eCG oneshot ($20mg{\times}6/300IU$) given intramuscularly 72 h prior to LOPU. For these groups, the number of follicles (mean ${\pm}$ SEM) observed which developed to larger than 2 mm in diameter were $1.6{\pm}2.5$, $4.3{\pm}3.1$, $5.5{\pm}4.2$, $6.6{\pm}2.1$ and $8.8{\pm}7.8$, respectively. Oocytes were aspirated by using OPU needles and a vacuum pump. The overall oocyte retrieval rates were 41.4%. Oocytes were matured in TCM-199 supplemented with 10% (w/v) bovine serum albumin + $10{\mu}g/ml$ FSH + $1{\mu}g/ml$ $17{\beta}$-estradiol for 27 h at $39^{\circ}C$ in 5% $CO_2$ in air. Oocytes were parthenogenetically activated by ionomycin combined with 6-diethylaminopurine (6-DMAP). Total oocyte maturation and cleavage rate were 67.3% and 78.8%, respectively. In summary, LOPU is a useful oocyte collection method in Korean black goats that can provide immature oocytes for transgenesis or nuclear transfer.

Effect of oocyte chromatin status in porcine follicles on the embryo development in vitro

  • Lee, Joo Bin;Lee, Min Gu;Lin, Tao;Shin, Hyeon Yeong;Lee, Jae Eun;Kang, Jung Won;Jin, Dong-Il
    • Asian-Australasian Journal of Animal Sciences
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    • 제32권7호
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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.

Reproductive Biology of the Temperate Soft Coral Dendronephthya suensoni (Alcyonacea: Nephtheidae)

  • Choi, Eun-Ji;Song, Jun-Im
    • Animal cells and systems
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    • 제11권2호
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    • pp.215-225
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    • 2007
  • The azooxanthellate soft coral Dendronephthya suensoni (Holem, 1895) is distributed mainly around Jejudo Island, Korea. This species was determined as gonochoric with a sex ratio of 2:1 (female:male). Both female and male colonies have one gametogenic cycle a year. The annual reproductive cycle of D. suensoni is dependent on the seawater temperature. In particular, reproduction of the male colony showed a higher positive correlation between seawater temperature and the mean diameter of the spermaries. Gametogenesis in females and males took 6 months and 12 months, respectively. The mean diameter of a mature oocyte was $249.29\;{\pm}\;36.24\;{\mu}m$, with a maximum size of $354.45\;{\mu}m$. Spawning could have occurred in the fall after the seawater temperature began to decrease.

돼지의 난포액 내 단백질인자의 탐색과 변화에 관한 연구 (Studies on Investigation and Change of Protein Factors in Porcine Follicular Fluids)

  • 지미란;정희태;양부근;이채식;박춘근
    • Reproductive and Developmental Biology
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    • 제34권3호
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    • pp.217-221
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    • 2010
  • When fully grown oocytes are removed from their follicles, they can resume meiosis and mature spontaneously under in vitro conditions. However, nuclear maturation under in vitro condition is not accompanied by complete cytoplasmic maturation, which is essential for successful fertilization and the initiation of zygotic development. This study analyzed change of proteins in follicular fluids during the porcine follicular development. Follicular fluids were collected from follicles of diameter 1~2 mm, 2~6 mm and 6~10 mm in ovary of slaughtered pigs. Total proteins were extracted from follicular fluids by M-PER Mammalian Protein Extraction Reagent. We confirmed totally 27 same spots, 1 spot from follicle fluid of 2~6 mm follicle and 5 spots from follicle fluid of 6~10 mm in diameter were analyzed by MALDI mass spectrometry and searched on NCBInr. In results, spot No. 28 from 2~6 mm follicle was Ig lambda chain C region, and spot No.32 and 33 from 6~10 mm was Apolipoprotein A-(APOA4). Spot No.29 and 31 were failed to analyze. These results indicate that the porcine oocyte during in vitro maturation depend on specific different expressed proteins may play an important roles in the sequence of molecular events in porcine oocyte maturation and follicular development.

볼락 (Sebastes inermis) 난병 유사구조의 전자현미경적 기재 (Electron Microscopical Description on the Egg Stalk-like Structure of the Rockfish, Sebastes inermis (Teleostei: Scorpaenidae))

  • 이정식;장영진
    • 한국수산과학회지
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    • 제35권2호
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    • pp.130-134
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    • 2002
  • 볼락의 난소는 낭상형이며, 다수의 난소소엽으로 구성되어 있었다. 난황축적 이전의 난모세포들은 난소소엽의 결체성 조직에 매몰되어 관찰되었다. 난모세포는 성장하면서 난소강으로 돌출되어 난병 유사구조를 가지고 포도송이와 같은 구조를 나타냈다 난소 소엽내에 매몰되어있는 직경 약 20$\mu$m 크기의 초기 난모세포에서 난황과립과 방사대는 관찰되지 않았다 직경 약 80$\mu$m의 초기 난모세포의 난병 유사구조 세포질에서는 전자밀도가 높은 난황과려들이 다수 관찰되었으나, 방사대의 pore canal system에서 음세포작용은 관찰되지 않았다. 이 시기에 난병 유사구조에서는 미세섬유, 발달된 소포체와 미토콘드리아, 전자밀도가 낮은 과립상 물질 그리고 활성화된 multivesicular body들이 관찰되었다. 이러한 결과들은 볼락의 난병 유사구조는 난모세포의 부착과 초기 난모세포의 외재적 난황축적에 관여함을 시사하는 것이라고 할 수 있다.