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  • Title/Summary/Keyword: Embryonic developmental

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Hatching of mouse balstocysts on somatic cell culture

  • Nah, Hee-Young;Gye, Myung-Chan
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 1998.07a
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    • pp.43-44
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    • 1998
  • Effect of somatic cell coculture on hatching of mouse blastocyst was examined. Mid-blastocysts were cocultured with granulosa cell primary culture or Sertoli cell line (TM4) derived from mouse testis for 48 hr. Blastocysts cultured in medium (10% FBS) started to hatch more faster than cocultured embryos during 12 hr of coculture. After then blastocysts cocultured with somatic cell hatched faster than control. Degeneration of embryos was also greately reduced by coculture. This result suggested the potentiation of hatching as well as embryonic viability by coculture with somatic cell and Sertoli cell line can be used for embryo coculture.

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The Making of a Competent Oocyte - A Review of Oocyte Development and Its Regulation

  • Tukur, Hammed A.;Aljumaah, Riyadh S.;Swelum, Ayman Abdel-Aziz;Alowaimer, Abdullah N.;Saadeldin, Islam M.
    • Journal of Animal Reproduction and Biotechnology
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    • v.35 no.1
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    • pp.2-11
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    • 2020
  • Assisted reproductive technologies (ART) merely depend on improving the oocyte maturation and their developmental competence to produce good quality embryos. Oocyte maturation passes through long and complex molecular steps starts from the early embryonic life and ends with sperm fertilization. Oocyte developmental competence can be attained by improving the nuclear and cytoplasmic mechanisms together with some epigenetic maturation. In this review, we highlight the cornerstones of oocyte maturation on both nuclear and cytoplasmic levels. Interfering or supporting these molecular mechanisms would help in the development of novel regulating agents for reproductive performance of humans and livestock species.

Assessements of Apoptosis in Bovine Embryos Reconstructed with Fetal Fibroblast

  • Lee, S. L.;Park, G.;S. Y. Choe
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.136-136
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    • 2003
  • Mainly due to deficiencies in nuclear reprogramming, gene expression and DNA fragmentation, which result in early and late embryonic losses, the overall success rate achieved by cloning techniques to date is low. This present study compared the incidences of DNA fragmentation during development of IVF, parthenotes (PT), nuclear transfer (NT) and transgenic (TG) embryos. Terminal deoxynucleotidyl transferase (TdT) nick-end labelling (TUNEL) with propidium iodide counter staining was used for determination of DNA fragmentation and total number, respectively. TG and NT donor cells were fetal fibroblasts with or without transfection with EGFP, and cultured in DMEM+15% FCS until confluent, for 5 days. At 19 h post-maturation (hpm), enucleated oocytes were reconstructed with donor cells and activated at 24 hpm with the combinations of ionomycin (5 M, 5 min) and cyclo-heximide (10 g/ml, 5 h) after electric fusion by a single DC pulse (1.6 KV/cm, 60 sec). Parthenotes were produced by the same activation protocol at 24 hpm. (중략)

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Follistatins have potential functional role in Porcine Embryogenesis

  • Kim, Dong-Hee;Chun, Ju Lan;Lee, Ji Hye;Kim, Keun Jung;Kim, Eun Young;Lee, Bo Myeong;Zhuang, Lili;Kim, Min Kyu
    • Korean Journal of Agricultural Science
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    • v.43 no.1
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    • pp.52-60
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    • 2016
  • In animal reproduction, the quality of oocytes and embryos has been evaluated by the expression of specific molecules. Follistatin (FST), which was isolated from follicular fluid, binds and bio-neutralizes the TGF-β superfamily members. Previous studies using the bovine model showed FST could be an important molecular determinant of embryo developmental competence. However, the effect of FST treatment on porcine embryo developmental competence has not been established. In this study, the effect of exogenous FST on porcine embryo developmental competence was investigated during in vitro culture. FST (10 ng/ml) treatment induced a significant decrease in the rate of cell arrest at the 4-cell stage. The expression levels of DNA-methyltransferase 1 (DNMT1), histone deacetylase 1 (HDAC1), and histone deacetylase 2 (HDAC2) were decreased in 4-cell stage embryos. FST treatment also resulted in significant improvements in developmental competence of embryos in terms of blastocyst formation rate and OCT-4 mRNA levels, the latter being related to pluripotency. In conclusion, during in vitro culture, FST treatment significantly ameliorated 4-cell block during embryonic development and improved embryo developmental competence. Therefore, FST treatment may potentially have a functional role in porcine embryogenesis that is broadly applicable to enhance in vitro embryo development.

Teratogenicity Evaluation of 2-Bromopropane Using Rat Whole Embryo Culture (랫드 전배아배양법을 이용한 2-Bromopropane의 최기형성 평가)

  • Kim Jong-Choon;Shin Dong-Ho;Kim Sung-Ho;Yang Young-Soo;Oh Ki-Seok;Jiang Cheng-Zhe;Chung Moon-Koo
    • Toxicological Research
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    • v.22 no.2
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    • pp.127-133
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    • 2006
  • Recently, we have reported that the environmental pollutant 2-bromopropane (2-BP) induces a significant embryo-fetal developmental toxicity in rats. However, the cause of developmental toxicity and the relationship between maternal and developmental toxicities could not be elucidated because the developmental toxicity of 2-BP was observed only in the presence of maternal toxicity The in vitro teratogenicity study using whole embryo culture was carried out to understand the teratogenic properties and the possible mechanism of teratogenicity induced by 2-BP in rats. Rat embryos aged 9.5 days were cultured in vitro for 48 hrs at medium concentrations of 0, 1, 3, or 10 mg/ml of 2-BP. Embryos were evaluated for growth, differentiation, and morphological alterations at the end of the culture period. At 10 mg/ml, 2-BP caused a delay in the growth and differentiation of embryos and an increase in the incidence of morphological alterations, including altered yolk sac circulation, abnormal axial rotation, craniofacial hypoplasia, open neuropore, absent optic vesicle and kinked somites. At 3 mg/ml, only a delay in the growth and differentiation of embryos was observed. There were no adverse effects on embryonic growth and development at the concentration of 1 mg/ml. The results showed that the exposure of 2-BP to rat embryos results in a developmental delay and morphological alterations at dose levels of 3 mg/ml culture media or higher and that 2-BP can induce a direct developmental toxicity in rat embryos.

Genomic Organization, Tissue Distribution and Developmental Expression of Glyceraldehyde 3-Phosphate Dehydrogenase Isoforms in Mud Loach Misgurnus mizolepis

  • Lee, Sang Yoon;Kim, Dong Soo;Nam, Yoon Kwon
    • Fisheries and Aquatic Sciences
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    • v.16 no.4
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    • pp.291-301
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    • 2013
  • The genomic organization, tissue distribution, and developmental expression of two paralogous GAPDH isoforms were characterized in the mud loach Misgurnus mizolepis (Cypriniformes). The mud loach gapdh isoform genes (mlgapdh-1 and mlgapdh-2) had different exon-intron organizations: 12 exons in mlgapdh-1 (spanning to 4.88 kb) and 11 in mlgapdh-2 (11.78 kb), including a non-translated exon 1 in each isoform. Southern blot hybridization suggested that the mud loach might possess the two copies of mlgapdh-1 and a single copy of mlgapdh-2. The mlgapdh-1 transcript levels are high in tissues requiring high energy flow, such as skeletal muscle and heart, whereas mlgapdh-2 is expressed abundantly in the brain. Both isoforms are differentially regulated during embryonic and larval development, during which their expression is upregulated with the progress of development. Lipopolysaccharide challenge preferentially induced mlgapdh-2 transcripts in the liver. Therefore, the two isoforms have diversified functionally; mlgapdh-1 is associated more closely with energy metabolism, while mlgapdh-2 is related more to stress/immune responses, in the mud loach.

Morphological Study on the Correlation of Prenatal and Postnatal Development between Mouse Parotid Salivary Gland and Tooth

  • Jeong, Soon-Jeong;Jeong, Moon-Jin
    • Applied Microscopy
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    • v.47 no.4
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    • pp.242-250
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    • 2017
  • The purpose of this study was to characterize the prenatal and postnatal development of the mouse parotid salivary gland and tooth, and to investigate the relationship between the developmental timing of the two organs. Development of parotid salivary gland begins on embryonic day 15 (E15), which is the prebud stage; E17 is the initial bud stage; E21 to postnatal day 3 (PN3) is the pseudoglandular stage; PN5 to PN10 is the canalicular stage; and PN21 is the terminal bud stage. At E15, the developing maxillary molar tissue is at the bud stage; at E17, it is at the cap stage; at E21, it is at the early bell stage; PN3 to PN5 comprises the advanced bell stage; at PN10, it is at the crown stage; at PN21, it is at the functional stage. Therefore, unlike the other major salivary glands, the development of mouse parotid salivary gland is completed through a process of prenatal and postnatal morphogenesis and becomes functional at about the same time as the developing tooth. The developmental completion times of the parotid salivary gland and tooth are closely related to the weaning time of animal.

Ruvbl1 is Essential for Ciliary Beating during Xenopus laevis Embryogenesis

  • Chan Young Kim;Hyun-Kyung Lee;Hongchan Lee;Hyun-Shik Lee
    • Development and Reproduction
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    • v.27 no.3
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    • pp.159-165
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    • 2023
  • The Ruvb-like AAA ATPase1 (Ruvbl1; also known as Pontin) is an evolutionary conserved protein belonging to the adenosine triphosphates associated with diverse cellular activities (AAA+) superfamily of ATPases. Ruvbl1 is a component of various protein supercomplexes and is involved in a variety of cellular activities, including chromatin remodeling, DNA damage repair, and mitotic spindle assembly however, the developmental significance of this protein is unknown and needs detailed investigation. We investigated the developmental significance of Ruvbl1 in multiciliated cells of the Xenopus laevis epidermis since ruvbl1 is expressed in the multiciliated cells and pronephros during X. laevis embryogenesis. The knockdown of ruvbl1 significantly impaired cilia-driven fluid flow and basal body polarity in the X. laevis epidermis compared to control embryos, but did not affect cilia morphology. Our results suggest that Ruvbl1 plays a significant role in embryonic development by regulating ciliary beating; however, further investigation is needed to determine the mechanisms involved.

Generation of Embryonic Stem Cell-derived Transgenic Mice by using Tetraploid Complementation

  • Park, Sun-Mi;Song, Sang-Jin;Choi, Ho-Jun;Uhm, Sang-Jun;Cho, Ssang-Goo;Lee, Hoon-Taek
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.121-121
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    • 2003
  • The standard protocol for the production of transgenic mouse from ES-injected embryo has to process via chimera producing and several times breeding steps, In contrast, tetraploid-ES cell complementation method allows the immediate generation of targeted murine mutants from genetically modified ES cell clones. The advantage of this advanced technique is a simple and efficient without chimeric intermediates. Recently, this method has been significantly improved through the discovery that ES cells derived from hybrid strains support the development of viable ES mice more efficiently than inbred ES cells do. Therefore, the objective of this study was to generate transgenic mice overexpressing human resistin gene by using tetrapioid-ES cell complementation method. Human resistin gene was amplified from human fetal liver cDNA library by PCR and cloned into pCR 2.1 TOPO T-vector and constructed in pCMV-Tag4C vector. Human resistin mammalian expression plasmid was transfected into D3-GL ES cells by lipofectamine 2000, and then after 8~10 days of transfection, the human resistin-expressing cells were selected with G418. In order to produce tetraploid embryos, blastomeres of diploid embryos at the two-cell stage were fused with two times of electric pulse using 60 V 30 μsec. (fusion rate : 93.5%) and cultured upto the blastocyst stage (development rate : 94.6%). The 15~20 previously G418-selected ES cells were injected into tetraploid blastocysts, and then transferred into the uterus of E2.5d pseudopregnant recipient mice. To investigate the gestation progress, two El9.5d fetus were recovered by Casarean section and one fetus was confirmed to contain human resistin gene by genomic DNA-PCR. Therefore, this finding demonstrates that tetraploid-ES mouse technology can be considered as a useful tool to produce transgenic mouse for the rapid analysis of gene function in vivo.

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Genetically Modified Human Embryonic Stem Cells Expressing Nurr1 and Their Differentiation into Tyrosine Hydroxylase Positive Cells in vitro.

  • Cho, Hwang-Yoon;Lee, Chang-Hyun;Kil, Kwang-Soo;Yoon, Ji-Yeon;Shin, Hyun-Ah;Lee, Gun-Soup;Lee, Young-Jae;Kim, Eun-Young;Park, SePill;Lim, Jin-Ho
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.104-104
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    • 2003
  • As an effort to direct differentiation of human embryonic stem (hES, MB03) cells to dopamine-producing neuronal cells, Nurr1 was transfected using conventional transfection protocol into MB03 and examined the expression of tyrosine hydroylase (TH) after differentiation induced by retinoic acid (RA) and ascorbic acid (AA). Experimentally, cells were transfected with linearized Nurr1 cDNA in pcDNA3.1 (+)-hygovernight followed by selection in medium containing hygromycin-B (150 μ/ml). Expression of Nurr1 mRNA was confirmed by RT-PCR and protein by immunocytochemistry in the drug resistant clones. In order to study the effect of Nurr1 protein on the differentiation pattern of ES cells, one of the positive clones (MBNr24) was allowed to form embryoid body (EB) for 2 days and were induced to differentiate for another 4 days using RA (1 μM) and AA (50 mM) (2-/4+ protocol) followed by selection in N2 medium for 10 or 20 days. After 10 days in N2 medium, cells immunoreactive to anti-GFAP, anti-TH, or anti-NF200 antibodies were 38.8%, 11%, and 20.5%, respectively. After 20 days in N2 medium, cells expressing GFAP, TH, or NF200 were 28%, 15% and 44.8%, respectively but approximately 9% of MB03 expressed TH protein when the cells were induced to differentiate using a similar prorocol, These results suggest that ectopic expression of Nurr1 enhances generation of TH+ cells as well as neuronal cells when hES cells were differentiated by 2-/4+ protocol.

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