• 제목/요약/키워드: Developmental competence

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Developmental competence of chimeric porcine embryos through the aggregation of parthenogenetic embryos and somatic cell nuclear transfer embryos

  • Joohyeong Lee;Lian Cai;Mirae Kim;Hyerin Choi;Dongjin Oh;Ali Jawad;Eunsong Lee;Sang-Hwan Hyun
    • 대한수의학회지
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    • 제63권1호
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    • pp.3.1-3.9
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    • 2023
  • The efficiency of somatic cell nuclear transfer (NT) in pigs is low and requires enhancement. We identified the most efficient method for zona pellucida (ZP) removal and blastomere aggregation in pigs and investigated whether the aggregation of NT and parthenogenetic activation (PA) of blastomeres could reduce embryonic apoptosis and improve the quality of NT-derived embryos by investigating. Embryonic developmental competence after ZP removal using acid Tyrode's solution or protease (pronase E). The embryonic developmental potential of NT-derived blastomeres was also investigated using well-of-the-well or phytohemagglutinin-L. We analyzed apoptosis in aggregate-derived blastocysts. The aggregation rate of protease-treated embryos was lower than that of Tyrode's solution-treated embryos (69.2% vs. 88.3%). No significant difference was observed between phytohemagglutinin-L and well-of-the-well (35.7%-38.5%). However, 2P1N showed a higher number of blastocysts compared to 3N (73.8% vs. 24.3%) and an increased blastocyst diameter compared to the control and 1P2N (274 ㎛ vs. 230-234 ㎛). In blastomeres aggregated using phytohemagglutinin-L, the apoptotic cell ratio was significantly higher in 1P2N and 3N than in 3P (5.91%-6.46% vs. 2.94%, respectively). Our results indicate that aggregation of one NT embryo with two PA embryos improved the rate of blastocysts with increased blastocyst diameter.

Impact of co-transfer of embryos produced by somatic cell nuclear transfer using two types of donor cells on pregnancy outcomes in dogs

  • Son, Young-Bum;Jeong, Yeon Ik;Jeong, Yeon Woo;Hossein, Mohammad Shamim;Hwang, Woo Suk
    • Animal Bioscience
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    • 제35권9호
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    • pp.1360-1366
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    • 2022
  • Objective: The present study analyzed the influence of co-transferring embryos with high and low cloning efficiencies produced via somatic cell nuclear transfer (SCNT) on pregnancy outcomes in dogs. Methods: Cloned dogs were produced by SCNT using donor cells derived from a Tibetan Mastiff (TM) and Toy Poodle (TP). The in vivo developmental capacity of cloned embryos was evaluated. The pregnancy and parturition rates were determined following single transfer of 284 fused oocytes into 21 surrogates and co-transfer of 47 fused oocytes into four surrogates. Results: When cloned embryos produced using a single type of donor cell were transferred into surrogates, the pregnancy and live birth rates were significantly higher following transfer of embryos produced using TP donor cells than following transfer of embryos produced using TM donor cells. Next, pregnancy and live birth rates were compared following single and co-transfer of these cloned embryos. The pregnancy and live birth rates were similar upon co-transfer of embryos and single transfer of embryos produced using TP donor cells but were significantly lower upon single transfer of embryos produced using TM donor cells. Furthermore, the parturition rate for TM dogs and the percentage of these dogs that remained alive until weaning was significantly higher upon co-transfer than upon single transfer of embryos. However, there was no difference between the two embryo transfer methods for TP dogs. The mean birth weight of cloned TM dogs was significantly higher upon single transfer than upon co-transfer of embryos. However, the body weight of TM dogs did not significantly differ between the two embryo transfer methods after day 5. Conclusion: For cloned embryos with a lower developmental competence, the parturition rate and percentage of dogs that remain alive until weaning are increased when they are co-transferred with cloned embryos with a greater developmental competence.

Improving the meiotic competence of small antral follicle-derived porcine oocytes by using dibutyryl-cAMP and melatonin

  • Jakree Jitjumnong;Pin-Chi Tang
    • Animal Bioscience
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    • 제37권6호
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    • pp.1007-1020
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    • 2024
  • Objective: We increased the nuclear maturation rate of antral follicle derived oocytes by using a pre-in vitro maturation (IVM) culture system and improved the developmental potential of these porcine pathenotes by supplementing with melatonin. Furthermore, we investigated the expression patterns of genes involved in cumulus expansion (HAS2, PTGS2, TNFAIP6, and PTX3) derived from small and medium antral follicles before and after oocyte maturation. Methods: Only the cumulus oocyte-complexes (COCs) derived from small antral follicles were induced with [Pre-SF(+)hCG] or without [Pre-SF(-)hCG] the addition of human chorionic gonadotropin (hCG) during the last 7 h of the pre-IVM period before undergoing the regular culture system. The mature oocytes were investigated on embryonic development after parthenogenetic activation (PA). Melatonin (10-7 M) was supplemented during in vitro culture (IVC) to improve the developmental potential of these porcine pathenotes. Results: A pre-IVM culture system with hCG added during the last 7 h of the pre-IVM period [Pre-SF(+)hCG] effectively supported small antral follicle-derived oocytes and increased their nuclear maturation rate. The oocytes derived from medium antral follicles exhibited the highest nuclear maturation rate in a regular culture system. Compared with oocytes cultured in a regular culture system, those cultured in the pre-IVM culture system exhibited considerable overexpression of HAS2, PTGS2, and TNFAIP6. Porcine embryos treated with melatonin during IVC exhibited markedly improved quality and developmental competence after PA. Notably, melatonin supplementation during the IVM period can reduce and increase the levels of intracellular reactive oxygen species (ROS) and glutathione (GSH), respectively. Conclusion: Our findings indicate that the Pre-SF(+)hCG culture system increases the nuclear maturation rate of small antral follicle-derived oocytes and the expression of genes involved in cumulus expansion. Melatonin supplementation during IVC may improve the quality and increase the blastocyst formation rate of porcine embryos. In addition, it can reduce and increase the levels of ROS and GSH, respectively, in mature oocytes, thus affecting subsequent embryos.

아토피 피부염 아동의 문제행동, 자아 역량 인식, 사회적 능력과 양육 스트레스와의 관계 (Behavioral Problem, Self-Perceived Competence, Social Competence, and Parental Stress in Children with Atopic Dermatitis)

  • 조복희;오세영;정자용;이형민
    • 대한가정학회지
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    • 제46권5호
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    • pp.97-109
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    • 2008
  • Atopic dermatitis is the fastest growing skin disorder among children from infancy to adolescence in Korea. The side-effects of atopic dermatitis not only include physical discomfort, but also psychological trauma which ultimately affects the developmental growth of children. This study was conducted in order to investigate the behavioral characteristics of an atopic dermatitis sufferer. As part of this research, the relative influence of behavioral problems and, self-perceived competence were analyzed in relation to the social competence of an atopic child. In total, 301 atopic and non-atopic children, between 2 and 6 years of age, and their mothers and teachers participated in the study. From this number, 109 children had atopic dermatitis, while 192 children did not. Mothers were asked to complete a parent-report questionnaire that required information on parental stress, according to the scale parenting methodology of Abbdin(1990) and Cho(1999). Teachers were subjected to teacher-report questionnaires which included topics on social competence, socia-emotional assessment and behavioral problems of an atopic child. Furthermore, children also completed questionnaires on self-perceived competence. According to analysis on K-CBCL, ITSEA, social competence, self-perceived competence and parental stress, children with atopic dermatitis showed higher scores in depression/anxiety and depression/withdrawal, compared to children in the normal control group. In relation to parental stress, daily stress, parental role stress, and stress related disease, mothers with atopic dermatitis children exhibited higher scores. In establishing relationships among the related variables, atopic children who demonstrated more social competence were more likely to suffer less from withdrawal, attention problems, and depression/anxiety. Stress related disease in mothers with atopic children was positively related to attention problems of the child. In terms of relative influences, behavioral problems was the most significant variable, accounting for 23% of variance. Lower behavioral problems was positively related to more social competence. In summation, this study investigated the general characteristics of atopic children. In conclusion, atopic children and their mothers had difficulty in dealing with this disease. It is our belief that an atopic child would not only require physical treatment, but also need appropriate psychological care.

Developmental Competence of Porcine NT Embryos Constructed by Microinjection of Fibroblast Cells into Vitrified Porcine Oocytes

  • Kim, Y.H.;Seok, H.B.;Kim, S.K.
    • 한국수정란이식학회지
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    • 제22권4호
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    • pp.265-269
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    • 2007
  • This study was conducted to investigate the efficacy of vitrification procedure for the cryopreservation of porcine oocytes and the utilization of vitrified oocytes as recipient cytoplasts for somatic cell nuclear transfer (NT), and observed that porcine oocytes are evaluated by pronuclear formation, and parthenogenetic development. Single fetal donor cells were deposited into the perivitelline space of vitrified enucleation oocytes, followed by electrical fusion and activation. NT embryos were cultured in NCSU-23 medium supplemented with 5% FBS, at $38.5^{\circ}C$ in 5% $CO_2$ and air. 1. When the developmental rates of the oocytes after being culture for $0{\sim}10$ hours vitrified with EDS and ETS were 42.0%, 38.0%, respectively. This results were lower than the control group(62.2%). 2. When the developmental rates of the oocytes after being culture for $0{\sim}10$ hours vitrified-thawed with sucrose and glucose, 5% PVP, NCSU-23 supplemented with 10% FBS were 33.3%, 25.9%, respectively. This results were lower than the control group(55.6%). 3. The fusion and development to the blastocyst stage between the NT embryos constructed with the vitrified and non-vitrified oocytes were significant differences. Developmental rate of oocytes and NT embryos constructed with the vitrified or non-vitrified oocytes were $13.0{\pm}2.4%\;and\;23.2{\pm}2.4%$, respectively.

대학과 협력한 초등수학 교사학습공동체의 발달 과정에 관한 연구 (A Study on the Development of Professional Learning Community in Mathematics Based on the Collaboration with University and Its Affiliated Elementary School)

  • 김남균
    • 한국수학교육학회지시리즈A:수학교육
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    • 제56권1호
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    • pp.119-130
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    • 2017
  • The purpose of this study is to explain the long term growth and development of elementary teachers' Professional Learning Communities(PLC) about mathematics implemented on an institutional basis. Especially, it is meaningful to analyze and present the development process and characteristics of PLC, which was started by the basis on the collaboration of a National University of Education and its affiliated elementary school. In this study, PLC activities during three years were analyzed according to the capacities and dimensions of a professional learning community. The developmental capacity of the PLC analyzed in this study can be summarized as follows. In the first year, development of organizational competence in terms of capacity, resources, structure, and system of exchanges was the main factor in personal competence, and the development of individual competence began to share collective learning and practice. In the second year, personal exchanges were active in all the topics of activities, and personal level competence was activated such that more activities of critical knowledge formation were performed on an individual level. On the basis of the development of the individual level formed in the second, individual competence and organizational capacity developed. Factors that have influenced the development of capacities of PLC include: disclosure of activities outside the community, participation in outsiders, provision of procedures to share equal participation and leadership, voluntary and critical participation of teachers, improvement of mathematics teaching methods, sharing themes and visions.

Porcine Oocytes with Meiotic Competence are Synchronized Early Stage of Germinal Vesicle (GV)

  • Park, Kwang-Wook
    • Reproductive and Developmental Biology
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    • 제34권3호
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    • pp.169-173
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    • 2010
  • Correlations between cumulus cells and germinal vesicle (GV) chromatin configuration were examined in porcine oocytes. Cumulus-oocyte complexes (COCs) were collected from 2~6 mm follicles and divided into three categories according to cumulus cell morphology. "A" group was compacted COCs with more than three cumulus cell layers. "B" group was COCs with less cumulus cell layers than "A" group. "C" group was COCs with one or less layer of cumulus cells. Cumulus cells were removed 0.1% hyaluronidase, and denuded oocytes were stained with Hoechst 33342. GV chromatin configuration was classified into GV-Con and GV-Dis. GV-Con meant that a nucleus was surrounded by condensed chromatin in a ring. GV-Dis meant that filamentous chromatin clumps were distributed in nucleus. The proportion (80.2%) of GV-Con in "A" group was significantly higher than "B" (62.0%) or "C" (44.9%). The proportion (55.1%) of GV-Dis in "C" group was significantly higher than "A" (19.8%) or "B" (38.0%). The meiotic competence of COCs was examined after 44 h culture. The proportion (90.0%) of oocytes reaching to metaphase II (M-II) in "A" group was significantly higher than "B" (76.5%) or "C" (45.5%). In conclusion, oocytes with good quality cumulus cell layers are synchronized early GV stage, and early GV stage is important for meiotic competence in pigs.

Impact of glycosylation on the unimpaired functions of the sperm

  • Cheon, Yong-Pil;Kim, Chung-Hoon
    • Clinical and Experimental Reproductive Medicine
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    • 제42권3호
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    • pp.77-85
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    • 2015
  • One of the key factors of early development is the specification of competence between the oocyte and the sperm, which occurs during gametogenesis. However, the starting point, growth, and maturation for acquiring competence during spermatogenesis and oogenesis in mammals are very different. Spermatogenesis includes spermiogenesis, but such a metamorphosis is not observed during oogenesis. Glycosylation, a ubiquitous modification, is a preliminary requisite for distribution of the structural and functional components of spermatids for metamorphosis. In addition, glycosylation using epididymal or female genital secretory glycans is an important process for the sperm maturation, the acquisition of the potential for fertilization, and the acceleration of early embryo development. However, nonemzymatic unexpected covalent bonding of a carbohydrate and malglycosylation can result in falling fertility rates as shown in the diabetic male. So far, glycosylation during spermatogenesis and the dynamics of the plasma membrane in the process of capacitation and fertilization have been evaluated, and a powerful role of glycosylation in spermatogenesis and early development is also suggested by structural bioinformatics, functional genomics, and functional proteomics. Further understanding of glycosylation is needed to provide a better understanding of fertilization and embryo development and for the development of new diagnostic and therapeutic tools for infertility.

Mitochondrial Porin Isoform AtVDAC1 Regulates the Competence of Arabidopsis thaliana to Agrobacterium-Mediated Genetic Transformation

  • Kwon, Tackmin
    • Molecules and Cells
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    • 제39권9호
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    • pp.705-713
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    • 2016
  • The efficiency of Agrobacterium-mediated transformation in plants depends on the virulence of Agrobacterium strains, the plant tissue culture conditions, and the susceptibility of host plants. Understanding the molecular interactions between Agrobacterium and host plant cells is crucial when manipulating the susceptibility of recalcitrant crop plants and protecting orchard trees from crown gall disease. It was discovered that Arabidopsis voltage-dependent anion channel 1 (atvdac1) mutant has drastic effects on Agrobacterium-mediated tumorigenesis and growth developmental phenotypes, and that these effects are dependent on a Ws-0 genetic background. Genetic complementation of Arabidopsis vdac1 mutants and yeast porin1-deficient strain with members of the AtVDAC gene family revealed that AtVDAC1 is required for Agrobacterium-mediated transformation, and there is weak functional redundancy between AtVDAC1 and AtVDAC3, which is independent of porin activity. Furthermore, atvdac1 mutants were deficient in transient and stable transformation by Agrobacterium, suggesting that AtVDAC1 is involved in the early stages of Agrobacterium infection prior to transferred-DNA (T-DNA) integration. Transgenic plants overexpressing AtVDAC1 not only complemented the phenotypes of the atvdac1 mutant, but also showed high efficiency of transient T-DNA gene expression; however, the efficiency of stable transformation was not affected. Moreover, the effect of phytohormone treatment on competence to Agrobacterium was compromised in atvdac1 mutants. These data indicate that AtVDAC1 regulates the competence of Arabidopsis to Agrobacterium infection.

돼지난포의 크기가 체외성숙-수정-배양에 의한 발육능력과 난자의 질에 미치는 영향

  • 안미현;김인덕;석호봉
    • 한국동물번식학회:학술대회논문집
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    • 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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    • pp.57-57
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    • 2003
  • 수정란의 체외생산(IVP)기술은 동물생산기술의 적용과 생리학이나 세포생물학의 기본 연구에 새로운 biotchnologies의 발생에 있어 매우 중요하고 흥미로운 사건으로서 여러 종류의 포유류에 적용되고 있다. 이러한 기술은 3가지의 절차를 밟아야 하는데 In vitro maturation(IVM), In vitro fertilization(IVF) 및 In vitro development(culture)(IVD)가 그것이다. 그런데 돼지는 다태동물로서 난소의 난포가 성장할 때 난포간 meiotic competence가 난자에 의하여 진행되므로 난포내의 난포액에 의하여 난자의 발육이 좌우된다고 보고 있다 돼지WP에 사용되는 난자는 난포의 직경이 3~5mm에서 수집하고 cumulus-oocyte complexes(COCs)의 형태에 따라 선택하는 것이 보편화되어 있다. 실험목적은 돼지 난소의 antral follicles이 성장할 때 크기에 의하여 oocyte meiotic competence의 방출에 어떤 영향을 주는지를 난자의 체외성숙과 체외수정 및 체외발육 비율을 각각 조사하여 미성숙 난자의 체외배양 기술을 개선할 목적이었다. 수행내용은 도축돈 난소의 난포크기별 3mm<, 3~5mm, 5mm)로 나누어, 다시 말해서 preantral stage, antral stage, postantral stage으로 구분하여 채취한 COCs를 IVM, IVF, IVC 상태에서의 COCs 형태, cell cleaved rate, developmental rate 등을 조사하였다.

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