• Title/Summary/Keyword: 난자형성과정

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Histological Studies on Gonad and Germ Cell Development of Diploid and Triploid Mud Loach (Misgurnus mizolepis) (2배체와 3배체 미꾸라지(Misgurnus mizolepis)의 생식소 발달)

  • Kim Bong-Seok;An Cheul Min;Kim Dong Soo
    • Journal of Aquaculture
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    • v.8 no.4
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    • pp.327-341
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    • 1995
  • A Study was conducted by using a histological method to examine gonadal development in diploid and triploid mud loach, Misgurnus mizolepis. Shape of ovaries from both groups looked like a single mass however, size of triploid ovary was significantly smaller than that of diploid ovary. Testis of triploid was also similar that of diploid in shape, but it was smaller than that of diploid. Ovarian development and oogenesis in diploid were significantly more rapid than those process in triploid, and first matured eggs were observed 100-days after hatching (SL, 5.68cm). Triploid ovary from hatching to 9-month-old had external appearance of undeveloped gonad and a few of oocytes of perinucleolus stage. However, normal matured eggs like those of female diploid were observed in one 5year-old triploid fish ovary examined. Although testicular development and sperrnatogenesis of diploid were prosperous sexually, testicular development of triploid were rather retarded compare to their male diploid counterpart in spermatogenesis.

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Identification of Oocyte-Specific Diva-Associated Proteins using Mass Spectrometry (Mass Spectrometry를 이용한 난자 특이적인 Diva와 상호작용하는 단백질의 동정)

  • Yoon, Se-Jin;Kim, Jung-Woong;Choi, Kyung-Hee;Lee, Sook-Hwan;Lee, Kyung-Ah
    • Clinical and Experimental Reproductive Medicine
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    • v.33 no.3
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    • pp.189-198
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    • 2006
  • Objective: We previously described that Diva is highly expressed in matured metaphase II (MII) oocytes compared to immature germinal vesicle (GV) oocytes in mouse. We report here that the expression of Diva transcript as well as protein is oocyte-specific. To elucidate its physiological role in oocyte, the binding partner(s) of Diva has been identified by using immunoprecipitation (IP) followed by Mass Spectrometry. Methods: NIH/3T3 cells were transiently transfected for 24 h with either empty vector for control or FLAG-tagged mouse Diva construct, and IP was performed with anti-FLAG antibody. The immuno-isolated complexes were resolved by SDS-PAGE on a 12% gel followed by Coomassie Blue staining. For in-gel digestion, 15 bands of interest were excised manually and digested with trypsin. All mass spectra were acquired at a positive reflector mode by a 4700 Proteomics Analyzer (Applied Biosystems, Framingham, MA). Proteins were identified by searching the NCBI nonredundant database using MASCOT Peptide Mass Fingerprint software (Matrixscience, London). Results: Diva-associated complexes were formed in FLAG-tagged mouse Diva-overexpressed NIH/3T3 cells via IP using anti-FLAG-conjugated beads. Among the excised 15 bands, actin and actin-binding proteins such as tropomyosin, tropomodulin 3, and ${\alpha}$-actinin were identified. Binding between Diva and actin or tropomyosin was confirmed by IP followed by Western blot analysis. Both bindings were also detected endogenously in mouse ovaries, indicating that Diva works with actin and tropomyosin. Conclusions: This is the first report that immuno-isolated Diva-associated complexes are related to actin filament of the cytoskeletal system. When we consider the association of Diva with actin and tropomyosin, oocyte-specific Diva may play a role in modulating the cytoskeletal system during oocyte maturation.

User-centric Context Model for Context-aware Application (맥락 인식 애플리케이션을 위한 사용자 중심의 맥락 모델)

  • Hong, Dong-Pyo;Woo, Woon-Tack
    • Proceedings of the Korea Information Processing Society Conference
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    • 2007.05a
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    • pp.810-813
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    • 2007
  • 본 논문에서는 맥락 인식 (context-aware) 애플리케이션 개발에 있어서 사용자의 맥락 정보를 보다 효과적으로 처리할 수 있는 사용자 중심의 맥락 모델을 제안한다. 유비쿼터스 컴퓨팅 개념의 확산과 함께 맥락 인식 애플리케이션들에 관한 많은 연구가 진행되고 있다. 하지만, 맥락에 대한 정의는 여전히 모호하며, 애플리케이션들 마다 서로 다른 형태의 맥락을 활용하고 있기 때문에, 맥락에 대한 보다 구체적인 정의와 다양한 애플리케이션에 활용 가능한 형태의 맥락 모델이 필요하다. 제안된 사용자 중심의 맥락 모델에서는 사용자가 애플리케이션과 상호작용할 때 사용자의 직접적인 명령을 제외한 사용자와 관련된 정보를 맥락으로 정의한다. 또한, 제안된 사용자 중심의 맥락은 5W1H 형태로 구조화한 맥락요소 (ContextElement), 맥락 요소들을 편리하게 처리할 수 있는 연산자들을 포함하는 맥락 (Context), 그리고 단편적인 맥락 정보뿐만 아니라 기존의 맥락 정보까지도 활용할 수 있는 맥락메모리 (ContextMemory)로 구성된다. 특히, 다양한 센서들로부터 획득된 정보를 맥락 모델의 인터페이스를 통해서 맥락 인식 애플리케이션에서 활용할 수 있기 때문에, 서로 다른 맥락 인식 애플리케이션들을 개발함에 있어서도 동일한 맥락 모델을 사용할 수 있는 장점이 있다. 제안된 맥락 모델의 유용함을 보이기 위해서, 센서로부터 획득된 맥락 정보를 처리하는데 소요되는 시간을 측정하는 실험을 하였다. 따라서 제안된 사용자 중심의 맥락 모델은 사용자와 맥락 인식 애플리케이션간 자연스러운 상호작용을 지원할 것으로 기대된다.) kcal/mol의 생성활성화 에너지 감을 나타내었고, TGA로부터의 분해활성화 에너지는 각각 31.94, 30.84, 24.16 kcal/mol의 값을 나타내었다.로 감소되었다(35.2% vs. 77.4%; p<0.01). 실험 2에서 다양한 정자 농도에 의한 정자 침투율과 정상 수정률을 바탕으로 판단했을 때 $4.6{\times}10^6/ml$의 정자 농도가 다른 정자 농도에 비해 난구 세포부착 난자의 체외 수정에 적합한 것으로 나타났다. 체외 수정과정에서 난구 세포 부착된 상태로 수정된 난자는 나화 난자에 비해 유의적으로(p<0.05) 높은 분할률(48.8% vs. 58.9%), 배반포 형성률(11.0% vs. 22.8%)과 배반포 세포수$(22{\pm}2\;vs.\;29{\pm}2)$를 나타내었다. 본 연구의 결과로부터 돼지의 체외 수정과정에서 난구 세포의 존재는 정자 침투를 저해하지만 분할률, 배반포 형성률 및 배반포의 세포수를 증가시키는 것으로 사료된다.수의 유출입 지점에 온도센서를 부착하여 냉각수의 온도를 측정하고 냉각수의 공급량과 대기의 온도 등을 측정하여 대사열의 발생을 추정할 수 있었다. 동시에 이를 이용하여 유가배양시 기질을 공급하는 공정변수로 사용하였다 [8]. 생물학적인 폐수처리장치인 활성 슬러지법에서 미생물의 활성을 측정하는 방법은 아직 그다지 개발되어있지 않다. 본 연구에서는 슬러지의 주 구성원이 미생물인 점에 착안하여 침전시 슬러지층과 상등액의 온도차를 측정하여 대사열량의 발생량을 측정하고 슬러지의 활성을 측정할 수 있는 방법을 개발하였다.enin과

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Differentiation of Border Cells During Oogenesis in Drosophila melanogaster (노랑초파리 난자 형성과정 동안의 경계세포의 분화)

  • Gye, Myung-Chan;Cho, Kyoung-Sang;Lee, Chung-Choo
    • Development and Reproduction
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    • v.2 no.1
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    • pp.45-52
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    • 1998
  • An enhncer detector line(EDL) having P[1ArB] insertion in X chromosome with expression of reporter gene (lacZ) in the polar cells and border cell of egg chamber was established and used to monitor the differentiation and migration of border cells during the oogenesis of Drosophila. differentiation of border cell from the anterior polar follicle cells was evident in stage-9 egg chamber of EDL149 which was characterized by migration of columnar follicle cells toward posterior of egg chamber surrounding the oocyte. Migration of border cells was observed in the stage-9 and -10 egg chambers. \beta -galactosidase activities were rapidly increased during the first 4 days after eclosion, and it coincided with the timing of border cell differentiation in the ovary during adult life. Homozygote of EDL149 showed some retardation of border cell migration , resulting absence of migration of some border cells in the anterior part of egg chamber or delayed migration of some border cells in the stage-10 egg chamber. These results suggest that the P[1ArB] of EDL149 is inserted at the locus of the structural gene required for the border cell migration. In addition to the expression in egg chambers, lacZ expression was also detected in the meiotic germ cells of testis and antenna, suggesting the possible requirement of the trapped gene function in these organ. this EDL and enhancer trapped gene might be useful for the study of developmentally regulated cell migration.

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Reproductive Cycles of Moroco oxycephalus and M. lagowskii in Korea (한국산 버들치와 버들개의 생식 주기에 관한 연구)

  • Kang, Young-Jin;Min, Mi-Sook
    • Korean Journal of Ichthyology
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    • v.11 no.2
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    • pp.117-125
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    • 1999
  • We investigated the reproductive cycles of two freshwater fishes, Moroco oxycephalus and M. lagowskii, in Korea. Seasonal changes in gonadosomatic index (GSI) and gonads were investigated histologically from April 1998 to April 1999. The reproductive cycles of two species were not shown any differences. The reproductive cycle can be divided into 5 phases : phase I (spent phase), phase II (immature phase), phase III (early developing phase), phase IV (late developing phase), and phase V (ripe phase). In phase I, the gonads of two species began to lose distinctly their weights from mid April, and reached the lowest GSI in late July (phase II). In September, the GSI values of testis and ovary increased very slowly (phase III) and gonadal developments rested during the winter season (phase IV). In March, the GSI values of M. oxycephalus and M. lagowskii began to increase, and reached the maximum in April (phase V). From the cyclic changes in the GSI and histological analyses, the spawning period was between mid April and mid May.

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Histological Studies on the Gametogensis and Reproductive Cycle of the Hard Clam , Meretrix Iusoria (백합, Meretrix Iusoria의 생식세포형성과정 및 생식주기에 관한 조직학적 연구)

  • Lee, Ju-Ha
    • The Korean Journal of Malacology
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    • v.13 no.2
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    • pp.131-141
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    • 1997
  • 전북 김제시 심포에서 1994년 1월부터 12월까지 월별로 채집된 백합 Meretrix lusoria의 생식세포형성과정 및 생식주기를 , 조직학적으로 조사한 결과는 다음과 같다. 백합은 자웅이체이고 난생을 하며, 생식소는 내장낭의 간중장선의 하부로부터 족부의 근육부근까지 분포하며, 성숙되면 팽대되지만 방란, 방정 후에는 위축된다. 생식소는 많은 난자형성여포들과 정자형셩여포들로 구성되어 있다. 여포에는 호산성 세포와 미분화간충직들이 들어 있는데, 이들은 초기 생식세포의 영양물질로 제공되고 있으며, 생식소가 발달함에 따라 점차 소실된다. 초기활성기의 난원세포는 직경 10$\mu\textrm{m}$ 내외이며, 초기 난모세포는 난병을 여포벽에 부착한 채 성장을 하여 70$\mu\textrm{m}$ 내외의 완숙란으로 된다. 정원세포가 성장을 하여 정보세포, 정세포를 거쳐 변태를 마친 정자는, 정자형성소낭의 중앙 내강에 정자속을 형성한다. 방란, 방정을 마친 생식소는, 일부 미방출된 생식세포가 퇴화 흡수되면서 비활성기를 지나 이듬해 수온상승과 더불어 새로운 발달을 시작한다. 생식주기는 연속적인 5단계로 구분할 수 있는데, 초기 활성기(1-3월), 후기 활성디 (2-5월), 완숙기(4-8월), 부분 방출기(6-9월), 퇴화 및 비활성기(9-2월)로 구분할 수 있다. 산란은 6월 (22$^{\circ}C$)부터 9월 ($25^{\circ}C$)까지 지속되지만, 산란성기는 7월(27$^{\circ}C$)-8월(28$^{\circ}C$)이다. 생식소지수는 1월에 0.58을 나타내며 5월에 4.60으로 최대값을 나타낸 후, 6월부터 점차 감소하기 시작하여 12월과 1월에는 1.0미만에 머무른다.

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Mesodermal Formation and Patterning during Ascidian Embryogenesis (멍게 배발생 과정에서 중배엽 형성과 패턴화)

  • 김길중;니시다히로키
    • Development and Reproduction
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    • v.6 no.2
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    • pp.77-82
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    • 2002
  • In ascidians, a primitive chordate, maternal cytoplasmic factors and inductive interactions are involved in the specification of cell fates in early embryos. The larval structure of ascidians is relatively simple, and the major mesodermal tissues of the tadpole larva are notochord, muscle, and mesenchyme. Formation of muscle cells is a cell-autonomous process, and localized maternal macho-l mRNA specify muscle fate in the posterior marginal zone of the early embryo. In contrast, inductive influence from endoderm precursors plays important roles in the specification of notochord and mesenchyme fates. FGF-Ras-MAPK signaling is involved In the induction of both tissues. The difference in responsiveness of the posterior mesenchyme and anterior notochord precursors to FGF signaling is caused by the presence or absence of intrinsic factors that inherited from the posterior-vegetal egg cytoplasm, respectively. In these inductions, directed signal polarizes the induced cells and promotes asymmetric cell divisions to produce two daughter cells with distinct fates.

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Profiles of microRNAs in Mice Follicles According to Gonadotropins during in vitro Culture (생쥐 난포의 체외배양 중 생식샘자극호르몬에 따른 미세리보핵산 발현 양상)

  • Kim, Yong-Jin;Ku, Seung-Yup;Kim, Yoon-Young;Oh, Sun-Kyung;Kim, Seok-Hyun;Choi, Young-Min;Moon, Shin-Yong
    • Clinical and Experimental Reproductive Medicine
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    • v.36 no.4
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    • pp.265-274
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    • 2009
  • Objective: MicroRNAs (miR) are known to repress target genes at post-transcriptional level and play important roles in development and maturation of cell. However, the expression profiles of miR during ovarian follicle maturation have not been fully elucidated. Here, we designed this study to investigate the expression profiles of miR in oocytes and granulose cells (G-cells) after in vitro culture according to gonadotropins and adding hCG. Methods: Ovaries from 12-day-old mice (C57BL6) were removed and preantral follicles were isolated and cultured in $20\;{\mu}L$-drop of culture media with supplementation of either rFSH, rLH, or rFSH+rLH. After their full maturation, follicles were incubated with rhCG and rEGF. RNA was isolated from oocytes and G-cells, and real-time PCR were performed with primers of miR known to be expressed in the mouse ovary (mmu-miR-16, -miR-27a, -miR-126, -miR-721). Results: FSH+LH group showed the highest ovulation and MII rates among gonadotropin groups. The profiles of miRs in oocytes and G-cells differed according to gonadotropin groups and adding hCG. The profiles of miRs showed divergent changes between oocytes and G-cells. Conclusion: miR expression profiles are altered by gonadotropins and supplementation of hCG during in vitro maturation of murine follicles. Target gene study must be necessary to validate these findings.

Studies on Mammalian Homolog and Flanking Sequence of Mouse MT Transposon-like Element, Clone MTi7(MTi7) (생쥐의 MT Transposon-like Element, Clone MTi7(MTi7) 유전자의 포유류 Homolog 및 Flanking Sequence에 대한 연구)

  • Kim Young-Hoon;Ko Minsu;Woo Dae-Gyun;Choi Donchan;Lee Kyung-Ah
    • Development and Reproduction
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    • v.7 no.2
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    • pp.119-126
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    • 2003
  • In the previous study, we obtained list of differentially expressed genes between postnatal day 1 and day 5 mouse ovaries using suppression subtractive hybridization(SSH) and found that MT Oansposon-like element, clone MTi7(MTi7) was one of the highly expressed genes in the day 5 mouse ovary(Park et at., 2002). Results of in situ hybridization and RNA interference revealed that the expression of MTi7 is oocyte-specific in the ovary and may be involved in the regulation of oocyte maturation(Park et at., 2003). At present, MTi7 sequence has been known only in the mouse. Therefore, the present study was accomplished 1) to identify MTi7 sequence in the other mammalian species, such as bovine, porcine, rat, and human, and 2) to evaluate the flanking sequence of the mouse MTi7 since it has transposon characteristics. Using ovarian cDNAs derived from low different species, we cloned and identified new MTi7 sequence showing a high degree of sequence homology with the mouse MTi7(87∼98%). By using inverse PCR, we found that the mouse MTi7 may intercalated the beta-carotene 15, 15'-monooxygenase(Bcdo) gene and/or serine protease inhibitor, Kunitz Dpe I(Spint 1) gene. By finding the MTi7 sequences in the other mammalian species and the flanking gene of the MTi7 in mouse, it is expected to reveal the role(s) of MTi7 in the oogenesis as well as folliculogenesis in the near future.

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Effects of Recipient Oocytes and Electric Stimulation Condition on In Vitro Development of Cloned Embryos after Interspecies Nuclear Transfer with Caprine Somatic Cell (수핵난자와 전기적 융합조건이 산양의 이종간 복제수정란의 체외발달에 미치는 영향)

  • 이명열;박희성
    • Reproductive and Developmental Biology
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    • v.28 no.1
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    • pp.21-27
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    • 2004
  • This study was conducted to investigate the developmental ability of caprine embryos after somatic cell interspecies nuclear transfer. Recipient bovine and porcine oocytes were obtained from slaughterhouse and were matured in vitro according to established protocols. Donor cells were obtained from an ear-skin biopsy of a caprine, digested with 0.25% trypsin-EDTA in PBS and primary fibroblast cultures were established in TCM-199 with 10% FBS. The matured oocytes were dipped in D-PBS plus 10% FBS + 7.5 $\mu$ g/ml cytochalasin B and 0.05M sucrose. Enucleation were accomplished by aspirating the first polar body and partial cytoplasm which containing metaphase II chromosomes using a micropipette with an out diameter of 20∼30 $\mu$m. A Single donor cell was individually transferred into the perivitelline space of each enucleated oocyte. The reconstructed oocytes were electric fusion with 0.3M mannitol fusion medium. After the electrofusion, embryos were activated by electric stimulation. Interspecies nuclear transfer embryos with bovine cytoplasts were cultured in TCM-199 medium supplemented with 10% FBS including bovine oviduct epithelial cells for 7∼9 day. And porcine cytoplasts were cultured in NCSU-23 medium supplemented with 10% FBS for 6 ∼8 day at $39^{\circ}C, 5% CO_2 $in air. Interspecies nuclear transfer by recipient bovine oocytes were fused with electric length 1.95 kv/cm and 2.10 kv/cm. There was no significant difference between two electric length in fusion rate(47.7 and 44.6%) and in cleavage rate(41.9 and 54.5%). Using electric length 1.95 kv/cm and 2.10 kv/cm in caprine-porcine NT oocytes, there was also no significant difference between two treatments in fusion rate(51.3 and 46.1%) and in cleavage rate(75.0 and 84.9%). The caprine-bovine NT oocytes fusion rate was lower(P<0.05) in 1 pulse for 60 $\mu$sec(19.3%), than those from 1 pulse for 30 $\mu$sec(50.8%) and 2 pulse for 30 $\mu$sec(31.0%). The cleavage rate was higher(P<0.05) in 1 pulse for 30 $\mu$sec(53.3%) and 2 pulse for 30 $\mu$sec(50.0%), than in 1 pulse for 60 $\mu$sec(18.2%). The caprine-porcine NT oocytes fusion rate was 48.1% in 1 pulse for 30 $\mu$sec, 45.2% in 2 pulse for 30 $\mu$sec and 48.6% in 1 pulse for 60 $\mu$sec. The cleavage rate was higher(P<0.05) in 1 pulse for 30 $\mu$sec(78.4%) and 1 pulse for 60 $\mu$sec(79.4%), than in 2 pulse for 30 $\mu$sec(53.6%). In caprine-bovine NT embryos, the developmental rate of morula and blastocyst stage embryos were 22.6% in interspecies nuclear transfer and 30.6% in parthenotes, which was no significant differed. The developmental rate of morula and blastocyst stage embryos with caprine-porcine NT embryos were lower(P<0.05) in interspecies nuclear transfer(5.1%) than parthenotes(37.4%).