• Title/Summary/Keyword: Parthenogenetic

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Parthenogenesis in Silkworm, Bombyx mori L.

  • Gangopadhyay Debnirmalya;Singh Ravindra;Kariappa B. K.;Dandin S. B.
    • International Journal of Industrial Entomology and Biomaterials
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    • v.10 no.1
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    • pp.1-10
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    • 2005
  • Parthenogenesis in mulberry silkworm, Bombyx mori L. acquires immense use in the development of outstanding homozygous lines with higher viability, hybrid vigour, combining ability and less phenotypic variability. It can serve as a powerful tool in controlling sex of the offsprings as well as a useful tool in selection. In fact India is the second largest silk producing country in the world next only to China and all the five types of natural silks viz., mulberry, oak tasar, tropical tasar, muga and eri are produced in India. However, little information is available on the role of artificial parthenogenesis in the development of superior silkworm breeds. This paper overviews some important studies carried out on artificial parthenogenesis, and outline of different types of parthenogenesis, methods of induction of artificial parthenogenesis, factors responsible for successful parthenogenetic development, cytogenetics of artificial parthenogenesis and role of artificial parthenogenesis in silkworm breeding. Besides, an attempt is made to describe briefly about parthenogenetic engineering which includes cloning in silkworm, artificial insemination, chimeras, hybridization, chromosomal substitution and recombinant DNA in silkworm.

Isolation and Characterization of Parthenogenetic Embryonic Stem (pES) Cells Containing Genetic Background of the Kunming Mouse Strain

  • Yu, Shu-Min;Yan, Xing-Rong;Chen, Dong-Mei;Cheng, Xiang;Dou, Zhong-Ying
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.1
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    • pp.37-44
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    • 2011
  • Parthenogenetic embryonic stem (pES) cells could provide a valuable model for research into genomic imprinting and X-linked diseases. In this study, pES cell lines were established from oocytes of hybrid offspring of Kunming and 129/Sv mice, and pluripotency of pES cells was evaluated. The pES cells maintained in the undifferentiated state for more than 50 passages had normal karyotypes with XX sex chromosomes and exhibited high activities of alkaline phosphatase (AKP) and telomerase. Meanwhile, these cells expressed ES cell molecular markers SSEA-1, Oct-4, Nanog, and GDF3 but not SSEA-3 detected by immunohistochemistry and RT-PCR. The pES cells could be differentiated into various types of cells from three germ layers in vitro by analysis of embryoid bodies (EBs) with immunohistochemistry and RT-PCR, and in vivo by observation of pES cell-derived teratoma sections. Therefore, the established pES cell lines contained all features of mouse ES cells. This work provides a new strategy for isolating pES cells from Kunming mice, and the pES cell lines could be applied as the cell model in research into genomic imprinting and epigenetic regulation of Kunming mice.

Existence of Amino Acids in Defined Culture Medium Influences In Vitro Development of Parthenogenetic and Nuclear Transfer Porcine Embryos

  • Won, Cheol-Hee;Park, Sang-Kyu;Kim, Ki-Young;Roh, Sang-Ho
    • Journal of Embryo Transfer
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    • v.23 no.4
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    • pp.245-250
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    • 2008
  • This study was designed to investigate the effect of essential amino acids (EAA) and/or non-essential amino acids (NEAA) on the development of parthenogenetic and somatic cell nuclear transfer (SCNT) porcine embryos in vitro. To evaluate the timing of amino acids supplementation, activated oocytes were cultured in NCSU23-PVA with EAA, NEAA or NEAA+EAA (AAs) during specific periods as below: EAA, NEAA or AAs were supplemented during Day 0 to 6 (whole culture period: ALL), Day 2 to Day 6 (post-maternal embryonic transition period: POST-MET), Day 5 to Day 6 (post-compaction period: POST-CMP), Day 0 to Day 2 (pre-maternal embryonic transition period: PRE-MET), or Day 0 to Day 4 (post-compaction period: PRE-CMP). Supplementation of NEAA decreased cleavage rates in PRE-MET and PRE-CMP and also decreased blastocyst rates in POST-CMP. On the other hand, EAA significantly enhanced blastocyst formation rate in POST-MET and no detrimental effect on embryonic development in other groups. Interestingly, NEAA and EAA had synergistic effect when they were supplemented to the medium during whole culture period. Supplementation of AAs also enhanced SCNT porcine embryo development whereas BSA-free medium without AAs could not supported blastocyst formation of SCNT embryos. In conclusion, existence of EAA and NEAA in defined culture medium variously influences the development of parthenogenetic and SCNT porcine embryos, and their positive effect are only occurred when both EAA and NEAA are supplemented to the medium during whole culture period. Additionally, AAs supplementation enhances the blastocyst formation of SCNT porcine embryos when they are cultured in the defined condition.

Effect of Parthenogenetic Mouse Embryonic Stem Cell (PmES) in the Mouse Model of Huntington′s Disease

  • 이창현;김용식;이영재;김은영;길광수;정길생;박세필;임진호
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.80-80
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    • 2003
  • Huntington's disease (HD) is an autosomal dominant neurodegenerative disorder characterized by motor, cognitive, and psychiatric symptoms, accompanied by marked cell death in the striatum and cortex. Stereotaxic injection of quinolinic acid (QA) into striatum results in a degeneration of GABAergic neurons and exhibits abnormal motor behaviors typical of the illness. The objective of this study was carried out to obtain basic information about whether parthenogenetic mouse embryonic stem (PmES) cells are suitable for cell replacement therapy of HD. To establish PmES cell lines, hybrid F1 (C57BL/6xCBA/N) mouse oocytes were treated with 7% ethanol for 5 min and cytochalasin-B for 4 hr to initiate spontaneous cleavage. Thus established PmES cells were induced to differentiate using bFGF (20ng/ml) followed by selection of neuronal precursor cells for 8 days in N2 medium. After selection, cells were expanded at the presence of bFGF (20 ng/ml) for another 6 days, then a final differentiation step in N2 medium for 7 days. To establish recipient animal models of HD, young adult mice (7 weeks age ICR mice) were lesioned unilaterally with a stereotaxic injection of QA (60 nM) into the striatum and the rotational behavior of the animals was tested using apomorphine (0.1mg/kg, IP) 7 days after the induction of lesion. Animals rotating more than 120 turns per hour were selected and the differentiated PmES cells (1$\times$10$^4$cells/ul) were implanted into striatum. Four weeks after the graft, immunohistochemical studies revealed the presence of cells reactive to anti-NeuN antibody. However, only a slight improvement of motor behavior was observed. By Nissl staining, cell mass resembling tumor was found at the graft site and near cortex which may explain the slight behavioral improvement. Detailed experiment on cell viability, differentiation and migration explanted in vivo is currently being studied.

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Optimization of Embryo Density and the Volume of Culture Medium for an Improvement of Mouse Parthenogenetic Embryo Development

  • Roh Sangho;Choi Young-Joo;Min Byung-Moo
    • Reproductive and Developmental Biology
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    • v.29 no.3
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    • pp.145-147
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    • 2005
  • Autocrine or paracrine mediators released by the early embryo are implicated in the support of embryonic development. Their mechanisms and optimal embryo density in the medium, however, are uncertain. This study was conducted to establish the optimal embryo density and culture medium volume in mouse parthenogenetic embryo culture. In experiment 1, culture of parthenogenetirally activated oocytes at a concentration of $2{\~}4$ embryos/${\mu}L$ significantly improved development to the blastoryst stage ($72{\%}{\leq}$) compared with culture at the lower ($0.2{\~}1$e mbryos/${\mu}L,\;0\~37.5\%$) and the higher ($5{\~}6$ embryos/${\mu}L,\;30\~53\%$) concentration for 120 h when the oocytes were cultured in a 5 ${\mu}L$ drop under mineral oil In experiment 2, the embryos cultured at a concentration of $2{\~}4$ embryos/${\mu}L$ in a 10 ${\mu}L$ drop ($81.1{\%}$) showed significantly higher blastocyst rates than those in a 5 ${\mu}L$ drop ($68.5{\%}$). This study optimizes in vitro culture condition by modifying embryo density and the volume of culture medium It may give appropriate level of autocrine and/or paracrine factors to enhance viability and subsequent normal development of mouse parthenogenetic embryos in vitro.

In Vitro Development and Chromosome Constitution of Porcine Parthenotes following Different Activation Treatments

  • Wi, Hae-Joo;Kwon, Dae-Jin;Park, Joo-Hee;Park, Choon-Keun;Yang, Boo-Keun;Cheong, Hee-Tae
    • Reproductive and Developmental Biology
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    • v.31 no.4
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    • pp.273-278
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    • 2007
  • This study was conducted to examine the protein kinase inhibitors, 6-dimethylaminopurine (DMAP) and cycloheximide (CHXM) on the development and chromosome constitution of porcine parthenogenetic embryos. In vitro matured oocytes were activated by electric stimuli (ES) or a combination of ES with culture in 2 mM DMAP or $10{\mu}g/ml$ CHXM for 4 hr. Activated oocytes were cultured in PZM-3 for 6 days. Some 1-cell embryos and blastocysts were fixed by air dry method to analyze the chromosome constitutions and/or total cell number. Blastocyst development of DMAP-treated group (26.7%) was significantly higher (p<0.05) than those of CHXM-treated and ES control groups. Ploidy in 1-cell stage embryos was not different among groups (77.3 to 81.0%), however, proportion of diploid chromosome constitutions was high in DMAP-treated group (61.9%, p<0.05). In the blastocyst stage, proportion of diploid chromosome plates was significantly high in DMAP-treated group (64.2%, p<0.05), and proportion of abnormal chromosome plates was higher in CHXM-treated group (36.6%, p<0.05) than DMAP-treated group (28.3%,). Proportion of embryos with abnormal chromosome constitutions was slightly increased by DMAP (40.0%) and CHXM (42.1%) treatment due to the increasing of mixoploid (47.4 and 52.0%). The present study shows that the DMAP treatment increase the development of porcine parthenotes. However, parthenogenetic activation by ES or combined treatment with ES and DMAP or CHXM detrimentally affects the chromosome constitutions of porcine parthenotes during early embryonic development, leads to increased abnormal ploidy in the blastocyst stage.