• 한국발생생물학회지:발생과생식
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• 제10권1호
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• pp.33-39
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• 2006

아무므불가사리의 생식소 발달과 생식주기를 밝히기 위하여 2003년 11월부터 2005년 2월까지 경상남도 고성 연안 해역에서 채집된 개체들을 대상으로 생식소 숙도지수 (GSI)의 월별변화, 생식소 발달과정 및 생식소 발달 단계별 난경 변화를 조사하였다. 생식소숙도지수의 월별 변화는 암컷과 수컷이 유사한 경향을 보였으며 암컷은 $3.88{\pm}3.04$, 수컷은 $0.87{\pm}0.57$의 값으로 3월에 연중 최대값을 가지다가 이후 서서히 감소하였다. GSI의 월별 변화와 생식소 발달의 조직학적 관찰을 근거로 생식 주기는 회복기($6{\sim}9$월), 성장기($10{\sim}1$월), 성숙기($2{\sim}3$월), 방출기($3{\sim}4$월), 퇴화 및 흡수기($4{\sim}5$월)의 연속적인 주기로 구분되었다. 아무르불가사리의 난발달 양상은 동시발달형이고 년 1회 산란하는 것으로 보인다.

• 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.

• 한국패류학회지
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• 제32권1호
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• pp.17-23
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• 2016

암컷 떡조개, Phacosoma japonicus의 난모세포 발달과 보조세포들과 관련된 난황형성과정의 미세구조적 연구를 위해 전자현미경적 관찰에 의해서 조사를 하였다. 난모세포들 내에서의 난황형성과정은 내인성 자율합성과정과 외인성 타가합성에 의해 일어나고 있다. 내인성 자율합성과정을 통해 일어나는 난황형성은 난모세포질 내의 골지복합체, 조면소포체 그리고 미토콘드리아가 결합된 작용에 의해서 일어난다. 그러나 외인성 타가합성과정은 초기난황형성난모세포들의 기저부로 들어오는 난소외부의 난황전구체물질들의 내포작용 (endocytosis)의 결합이 관여하고 있다. 본 연구에서 전난황형성난모세포들과 난황형성난모세포들에 부착된 보조세포들이 용이하게 관찰되었다. 특히, 보조세포들은 영양공급에 의해 전난황형성 난모세포들의 발달에 관여하며, 난황전구체의 엔도시토시스에 의해 초기 및 후기 난황형성 난모세포들 내에서 난황형성에 관여한다. 성숙난모세포에 부착된 보조세포들의 기능은 보조세포들의 세포질 내에서 난황형성을 위해 지질과립들과 글리코겐입자들을 축적한다.

• Clinical and Experimental Reproductive Medicine
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• 제13권2호
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• pp.121-127
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• 1986

We have reviewed 59 cases of patients amoung 65 cases who underwent IVF and ET with reasonable indications irom 1984 and the results as follows. 1. Major indications for IVF and ET were tubal factor (40.7%), unexplained infertility (25.4%), endometriosis (15.3%), failed AID and AIH (10.1 %), and sperm abnormality (8.5%). 2. For superovulation of human oocytes, l00mg of clomiphene citrate and 75 IU of HMG used. The monitoring of oocyte maturation was bone by ultrasound examination and serum 17-${\beta}$ estradiol, LH values. The peak $E_2$ value was 956.36${\pm}$702.13 pg/ml. 3. The oocytes were obtained by laparoscopy 24-36 hours after the injection of HCG. 4. The mean numbers of follicles at laparoscopy was 3.06 and the successful rate of laparoscopy was 79.7%. 5. And 165 follicles were aspirated from which 98 oocytes were recovered, 59.4% of all follicles had at least one oocyte aspirated. 21.4% of the eggs were mature, 52.0% were moderate, 26.5%. were immature. 6. 67.3% of oocytes were cleaved and were transferred at 4-6 cell stages. 7. Four pregnancies including one chemical pregnancy and one spontaneous abortion were established by ${\beta}$-subunit, u-hCG and ultrasound examinations.

• Clinical and Experimental Reproductive Medicine
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• 제40권4호
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• pp.148-154
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• 2013

Objective: To compare the mouse oocyte vitrification outcomes of the CryoLogic vitrification method (CVM) and the conventional open method using a Cryotop. Two CVM methods (original CVM and modified CVM) were tested. Methods: Mature oocytes obtained from female BDF-1 mice were vitrified by two-step exposure to equilibrium and vitrification solutions. Three vitrification protocols were tested on three groups: the CVM-kit, modified CVM, and Cryotop groups. After exposure to the two solutions, the oocytes were vitrified. After warming, the oocytes were fertilized in vitro, and the embryo development was assessed. Blastomeres positive for caspase were counted using an in situ assay kit. The spindle morphology and chromosome configurations of warmed vitrified oocytes were also assessed. Results: The modified CVM and Cryotop groups showed similar developmental capacities, and similar proportions of cells with intact spindles and chromosome configurations. The modified CVM protocol was superior to the original CVM protocol for developmental competence and intact spindle preservation. However, the CVM group showed a relatively higher number of apoptotic cells in blastocysts. Conclusion: Closed vitrification using the modified CVM protocol may be used as an alternative to the conventional open method, but strategies to decrease apoptosis in the blastomere need to be investigated.

• Molecules and Cells
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• 제40권8호
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• pp.558-566
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• 2017

Regular monitoring on experimental animal management found the fluctuation of ART outcome, which showed a necessity to explore whether superovulation treatment is responsible for such unexpected outcome. This study was subsequently conducted to examine whether superovulation treatment can preserve ultrastructural integrity and developmental competence of oocytes following oocyte activation and embryo culture. A randomized study using mouse model was designed and in vitro development (experiment 1), ultrastructural morphology (experiment 2) and functional integrity of the oocytes (experiment 3) retrieved after PMSG/hCG injection (superovulation group) or not (natural ovulation; control group) were evaluated. In experiment 1, more oocytes were retrieved following superovulation than following natural ovulation, but natural ovulation yielded higher (p < 0.0563) maturation rate than superovulation. The capacity of mature oocytes to form pronucleus and to develop into blastocysts in vitro was similar. In experiment 2, a notable (p < 0.0186) increase in mitochondrial deformity, characterized by the formation of vacuolated mitochondria, was detected in the superovulation group. Multivesicular body formation was also increased, whereas early endosome formation was significantly decreased. No obvious changes in other microorganelles, however, were detected, which included the formation and distribution of mitochondria, cortical granules, microvilli, and smooth and rough endoplasmic reticulum. In experiment 3, significant decreases in mitochondrial activity, ATP production and dextran uptake were detected in the superovulation group. In conclusion, superovulation treatment may change both maturational status and functional and ultrastuctural integrity of oocytes. Superovulation effect on preimplantation development can be discussed.

• Clinical and Experimental Reproductive Medicine
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• 제48권4호
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• pp.347-351
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• 2021

Objective: We investigated the impact of vitamin D₃ (VD3) supplementation during mouse preantral follicle culture in vitro and the mRNA expression of 25-hydroxylase (CYP2R1), 1-alpha-hydroxylase (CYP27B1), and vitamin D receptor (VDR) in mouse ovarian follicles at different stages. Methods: Preantral follicles were retrieved from 39 BDF1 mice (7-8 weeks old) and then cultured in vitro for 12 days under VD3 supplementation (0, 25, and 50 pg/mL). Follicular development and the final oocyte acquisition were assessed. Preantral follicles were retrieved from 15 other BDF1 mice (7-8 weeks old) and cultured without VD3 supplementation. Three stages of mouse ovarian follicles were obtained (preantral, antral, and ruptured follicles). Total RNA was extracted from the pooled cells (from 20 follicles at each stage), and then reverse transcriptase-polymerase chain reaction was performed to identify mRNA for CYP2R1, CYP27B1, and VDR. Results: The survival of preantral follicles, rates of antrum formation and ruptured follicles (per initiated follicle) and the number of total or mature oocytes were all comparable among the three groups. Both CYP2R1 and CYP27B1 were expressed in antral and ruptured follicles, but not in preantral follicles. VDR was expressed in all three follicular stages. Conclusion: VD3 supplementation in vitro (25 or 50 pg/mL) did not enhance mouse follicular development or final oocyte acquisition. Follicular stage-specific expression of CYP2R1, CYP27B1, and VDR was observed.

• Asian-Australasian Journal of Animal Sciences
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• 제20권8호
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• pp.1190-1195
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• 2007

This study was conducted to improve efficiency of a follicle culture system without reducing developmental competence of intrafollicular oocytes. Preantral follicles (100 to $125{\mu}m$ in diameter) of F1 hybrid (B6CBAF1) mice were cultured singly for 216 h in modified ${\alpha}$-MEM-glutamax medium, to which 2.5 IU/ml hCG and epidermal growth factor was added 16 h prior to the end of culture. Medium change was either performed three times (54 h interval), twice (72 h interval), once (108 h interval), or not at all (216 h interval). Maturation (progression to the metaphase II stage) of intrafollicular oocytes was detected from 4 days after culture in the three-times change treatment, while all treatments yielded mature oocytes from day 5 of culture. Compared with the three-times change, decreasing the change frequency to once did not reduce the capacity to begin maturation (germinal vesicle breakdown of 82 to 86%), to mature (78 to 79%) and to develop into blastocysts after parthenogenetic activation (29 to 32%). Morphological parameters were similar among these treatments. Except for the no medium change treatment, similar colony-forming activity of inner cell mass cells after culturing of blastocysts in leukemia inhibitory factor-containing medium was detected, while the morphology of the colony-forming cells deteriorated in the change-once treatment compared with the change twice or three-times. In conclusion, the efficiency of the preantral follicle culture system could be improved by reducing frequency of medium change up to a 72 h interval (three times in total 216 h culture) without decreasing developmental competence of oocytes.

• 한국발생생물학회지:발생과생식
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• 제4권2호
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• pp.175-180
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• 2000

양산 내원사 계곡에 서식하는 난태생인 다슬기(Semisulcospira libertina libertina)를 재료로 하여 번식주기를 조사하였다. 서식지 수온은 연중 1.3~22.5$^{\circ}C$였으며, 육중량비는 수온이 상승하기 시작하는 3월에 최고값을 보였고, 8월에 최저값을 나타내었다. 난모세포의 평균 직경은 7월에 249.6$\pm$2.6 $mu extrm{m}$로 최대값을 나타냈으며, 12월에 134.3$\pm$2.8 $\mu\textrm{m}$로 연중 최소값을 보였다. 다슬기는 혹서기와 혹한기를 제외하고 주년 번식하는 종으로서, 연간 2회의 주된 번식주기를 가져, 암컷에서 분열증식기 3월ㆍ10월, 성장기 4~5월ㆍ11월, 성숙기 6~7월ㆍ12월, 배란기 8월ㆍ1월, 출산기 9~10월ㆍ3~5월, 휴지기 9월ㆍ2월, 수컷에서는 분열증식기 3월ㆍ10월, 성장기 4월ㆍ11월, 성숙기 5~6월ㆍ12월, 교미기 7~8월ㆍ1월, 휴지기 9월ㆍ2월인 것으로 판정되었다.

• 한국수정란이식학회지
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• 제17권2호
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• pp.117-121
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• 2002

본 연구는 소형견의 불임 해결을 위해 미숙난자를 보존 후 이용할 수 있는지의 여부를 판명하기 위하여 미성숙 난포란을 시간별로 배양한 뒤 vitrification 동결 융해후 체외수정율과 생존율을 조사하였다. 1 미숙난포란을 회수 후 1, 6, 12, 24시간 성숙 배양 후vitrification동결 융해 후 체외수정 시켰을때 수정율은 각각 31.4％. 22.5％, 11.9％ 및 5.3％로서 대조군의 수정율 60.0％에 비해 낮은 성적이었고, 회수 후 시간이 경과되지 않은 난포란이 높은 체외수정율을 나타냈다. 2. 미숙난포란을 회수 후 1, 6, 12 및 24시간 배양시킨 난포란을 vitrification 동결 융해 후 체외 수정시킨 배의 생존율은 각각 55.0％, 40.0％, 28.6％ 및 17.1％로써 대조군의 76.7％에 비해 낮은 생존율을 나타냈다.