• 한국수정란이식학회지
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• 제15권3호
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• pp.287-293
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• 2000

This study was undertaken to access the effects of collection method, room temperature at oocyte recovery and culture media on the oocyte quality, fertilization and cleavage rates of in vitro matured and fertilized oocytes of Korean native goats. Ovaries obtained from a slaughterhouse were transported to the laboratory and were divided into 2 groups. One group of ovaries was maintained at 30 to 35$^{\circ}C$ of the room temperature and another group was remained at 20 to $25^{\circ}C$ during oocyte recovery. The oocytes were recovered by follicle aspiration, slicing and aspiration+slicing methods from 3 groups of follicles according to size; <2 mm, 2 to 6 mm and >6 mm. The matured oocytes were inseminated with buck epididymal spermatozoa at a concentration of 3~3.5$\times$10$^{6}$ m1 and fertilization was identified when 2 pronuclei were present in the cytoplasm. Although the recovery rate per ovary obtained by the combination of follicle aspiration + slicing(19.6$\pm$2.2) method was higher than aspiration(11.7$\pm$1.1) and slicing(14.8$\pm$1.8) collection, optimal recovery according to oocyte grades resulted form ovarian slicing compared to aspiration or combined methods(P<0.05). However, no significant differences were found in the mean number(2.5$\pm$1.8; 3.3$\pm$3.3; 2.9$\pm$2.4) and the proportion of favorable oocytes(Grades I, II and III) recovered(31.6%, 36.0%, 36.4%,) according to follicle size(<2 mm; 2 to 6 mm; >6 mm). Fertilization rate was 60.0%, 67.7%, 70.6% and 56.4% and the proportion of embryos/zygotes was 11.1%, 7.1%, 5.0% and 2.8% in 20~$25^{\circ}C$/BO, 30~35$^{\circ}C$/BO, 20~$25^{\circ}C$/TALP and 30~35$^{\circ}C$ /groups, respectively.

• Nutrition Research and Practice
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• 제16권3호
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• pp.314-329
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• 2022

BACKGROUND/OBJECTIVES: Oocyte lipid droplets play a crucial role in meiosis and embryo development. Biotin is associated with fatty acid synthesis and is the coenzyme for acetyl-CoA carboxylase (ACC). The effects of a biotin deficiency on the oocyte lipid metabolism remain unknown. This study examined the effects of a biotin deficiency and its replenishment on murine 1) oocyte lipid droplet levels, 2) ovary lipid metabolism, and 3) oocyte meiosis. MATERIALS/METHODS: Mice were divided into 3 groups: control, biotin deficient (BD), and recovery groups. The control and BD groups were fed a control diet or BD diet (0.004 or 0 g biotin/kg), respectively. The recovery group mice were fed a BD diet until day 21, and were then fed the control diet from days 22 to 64. This study then quantified the oocyte lipid droplet levels, assessed the oocyte mitochondrial function, and examined the ability of oocytes to undergo meiosis. Ovarian phosphorylated ACC (p-ACC), lipogenesis, β-oxidation, and ATP production-related genes were evaluated. RESULTS: The BD group showed a decrease in lipid droplets and mitochondrial membrane potential and increased p-ACC levels. In the recovery group, the hepatic biotin concentration, ovarian p-ACC levels, and mitochondrial membrane potential were restored to the control group levels. On the other hand, the quantity of lipid droplets in the recovery group was not restored to the control levels. Furthermore, the percentage of oocytes with meiotic abnormalities was higher in the recovery group than in the control group. CONCLUSIONS: A biotin deficiency reduced the oocyte lipid droplet levels by downregulating lipogenesis. The decreased lipid droplets and increased oocyte meiosis failure were not fully restored, even though the biotin nutrition status and gene expression of lipid metabolism was resumed. These results suggest that a biotin deficiency remains robust and can be long-lasting. Biotin might play a crucial role in maintaining the oocyte quality.

• 한국어병학회지
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• 제16권2호
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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.

• 한국패류학회지
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• 제25권2호
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• pp.145-151
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• 2009

담수 복족류인 다슬기, Semisulcospira libertina libertina의 난자형성과정 동안 생식세포의 미세구조 변화를 광학현미경과 전자현미경으로 관찰하였다. 난소는 나탑 후방부의 간췌장의 표면에 위치하며, 성숙시기에는 녹색을 나타내었다. 난소는 다수의 난자형성소낭들로 이루어져 있었다. 난자형성과정은 5단계로, 난원세포기, 난황형성전기, 난황형성개시기, 난황형성활성기, 성숙기로 구분하였다. 난원세포는 직경 $4-6\;{\mu}m$의 원형으로 커다란 핵을 가진다. 난황형성전기의 난모세포는 직경이 약 $20\;{\mu}m$이며, H-E 염색에서 세포질은 호염기성을 나타냈다. 난황형성개시기의 난모세포는 직경 $60-80\;{\mu}m$ 내외로 세포질에는 전자밀도가 낮은 난황과립들을 가지며, 난병에 의하여 난자형성소낭에 연결되어 있었다. 난황형성 활성기의 난모세포는 직경 $100-120\;{\mu}m$로 난황과립의 전자밀도, 크기, 양이 증가하였으며, 세포질에는 골지체, 미토콘드리아, 조면소포체들이 발달하였다. 성숙기 난모세포의 세포질 대부분은 전자밀도가 높은 단백질성의 난황과립들로 채워져 있었으며, 난막에서는 길이 약 $1.1\;{\mu}m$의 미세융모가 관찰되었다.

• 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.

• 한국발생생물학회지:발생과생식
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• 제16권3호
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• pp.185-193
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• 2012

For the gonadal sex management of younger longtooth grouper (Epinephelus bruneus), this work investigated the timing and histological process of ovary differentiation and oocyte development of longtooth grouper larvae and juvenile. Specimens (from 1 to 365 DAH) were collected for gonadal histological study from June 2008 to August 2009. Rearing water temperature was ranged from 20 to $24^{\circ}C$. The primordial germ cells could be observed from 10 to 15 DAH, while undifferentiated gonad occurs from 20 to 50 DAH in longtooth grouper. The initial ovarian phase was 60 to 110 DAH with the formation of ovarian cavity and the increased in size of gonad. The ovarian phase started at 140 DAH with appearance of oogonia. The gonad at 365 DAH appeared to have full of oogonia and primary growth stage oocyte. Formation of ovarian cavity indicates that the ovarian differentiation beginning at 60 DAH in longtooth grouper. The gonads in longtooth grouper differentiated directly into ovaries in all fish examined.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.51-51
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• 2003

Oct-4 is a maternally expressed octamer-binding protein encoded by the murine Oct-4 gene. It is present in unfertilized oocytes, but also in the inner cell mass and in primordial germ cells. In addition, Oct-4 is the first transcrition factor described that is specific for the blastocysts stage bovine embryos. The spatial and temporal expression patterns were further determined using Immunohistochemistry. With this technique Oct-4 protein expression is detected in the oocyte, in the blastocyst. After pregnancy Oct-4 expression is restricted ovary and placental tissue. Therefore Oct-4 is a transcription factor that is specifically expressed in cells participating in the pregnancy of Korean native cattle. These result suggest that Oct-4 localization and expression may contribute to the defects in the developmental normal seen in Korean native cattle.

• 한국수정란이식학회지
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• 제11권1호
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• pp.41-50
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• 1996

This experiment was carried out to investigate the ovarian responses of the ovulation point, ovarian weight and size, the number of ovarian follicles and collected embryos, and to study the effects of the developmental stages (oocytes, 2-4 cell. 8-16 cell and morulae), additional levels of Ficoll (0, 15, 30%) on the survival rate (FDA-test) of rat embryos frozen in vitrification solution (20% glycerol + 10% ethylene glycol + 10% sucrose). Sunanarized results was as follows; 1. The mean ovulation point per head was 7, and the weight of ovaries was 0.03g. The size of ovary was 5.9 mm(L) and 4.6 mm(W), and the number of ovarian follicles over and below 2 mm was 4.7 and 8.7, respectively. The number of the collected embryos per head was 5.5 (79%). 2. 2. The FDA score of embryos frozen in 20 G 10 E 10 S without Ficoll was 2.8 (oocyte), 2.6 (2-4 cell), 3.9 (8-16 cell) and 3.6 (morula), respectively. However, there were no significant differences among treatments. 3. The FDA score of embryos frozen in 20 G 10 E 10 S with 15% Ficoll was 3.4 (oocyte), 4.0 (2-4 cell), 4.7 (8-16 cell) and 4.8 (morulae), respectively (P>0.05). 4. The FDA score of embryos frozen in 20 G 10 E 10 S with 30 % Ficoll was 3.7 (oocyte), 3.2 (2-4 cell), 4.4 (8-16 cell) and 4.4 (morulae), respectively (P>0.05). 5. As shown in the above results, the higher survival rate was obtained in the treatment of 15% Ficoll than that of 30%. And the survival rate (FDA-test)of the oocytes and 2-4 cell stages of the rat embryos was lower than that of 8~16 cell and morulae stages. It was considered that 8-16 cell and morulae could be available for the successful freezing by vitrification of rat embryos with 15% Ficoll except for oocytes.

• 한국양식학회지
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• 제2권1호
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• pp.21-32
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• 1989

이스라엘 잉어 난모세포의 발달단계와 혈청중 호르몬 농도의 변화를 알기 위해서 2월부터 3월까지 본 연구를 수행했다. 본 연구에서 얻어진 결과는 다음과 같다. 1. 난소가 발달되는 3월에 들어와서 혈청중 LH 수준이 급격히 상승하기 시작했다. 2. LH 수준은 G.S.I.의 변화와 높은 상관관계에 있었다. 3. 3월경부터 G.S.I.가 급격하게 증가했다 4. 3월중에는 성숙단계가 상당히 증가했다. 5. Early Perinuclcolus oocyte가 late perinucleolus core로의 빠른 발달을 나타냈다. 6. 난황이 축적됨으로써 late perinucleolus oocyte가 초기성숙단계인 oocyte로 되는 시기에 vitellogenesis가 시작되었다. 7. Oocyte가 후기성숙단계에 도달했을 때 세포질내에 난황으로 가득 채워졌다. 8. 이스라엘 잉어 난소의 헌미경적 형태의 변화는 G.S.I. 외관상의 변화와 연관성이 깊었다 9. LH는 이스라엘 잉어의 성성숙에 중요한 역할을 수행하는 것으로 나타났다.

• 한국동물학회지
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• 제14권4호
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• pp.181-191
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• 1971

각시물자라의 卵巢小管의 관찰결과 두 개의 卵巢 중 各各 다섯 개의 端榮養室型의 卵巢小管을 가졌으며, 各 齡마다의 生殖室의 發生狀態는 다음과 같았다. 第4齡에 있어서 生殖室의 上端部位는 미분화된 細胞로 되어 있었으며, 中央部에는 球形에 가까운 榮養束이 있었다. 그리고 下端部位에는 卵細胞가 있었고 그 다음은 種을 거꾸로 세운 모양의 prefollicular tissue가 있었다. 그리고 榮養索은 아주 미발달된 狀態였다. 第5齡에 있어서 榮養束은 방추형이며 榮養束 아래에 위치하고 있는 卵細胞는 prefollicular tissue 內에서 榮養索으로 榮養束과 연결되어 있었다. 成蟲에 있어서는 生殖室 다음에 榮養室이 계속되었으며 榮養室內에는 發達과정에 있는 卵細胞로 가득차 있었다. 그리고 yolk granule은 濾胞上皮細胞層으로부터 卵細胞의 內部로 축적되어졌다. 그리고 나중에는 卵細胞質은 완전히 同質性이 되었다. 그러므로 齡이 進行함에 따라 榮養索이 卵細胞에서 卵殼이 생길 때까지 발달되었으며 濾胞細胞는 두 개의 核을 가진 狀態로 되었다. 그리고 成蟲의 卵巢小管에는 卵殼을 갖지않는 部分과 卵殼을 갖는 部分이 있었다.