• Title/Summary/Keyword: mouse embryo

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Expression patterns of Rho-associated protein kinase signaling pathway-related genes in mouse submandibular glands

  • Kim, Ki-Chul;Roh, Sangho
    • International Journal of Oral Biology
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    • v.46 no.2
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    • pp.81-84
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    • 2021
  • Salivary glands are exocrine glands that secrete saliva into the oral cavity, and secreted saliva plays essential roles in oral health. Therefore, maintaining the salivary glands in an intact state is required for proper production and secretion of saliva. To investigate a specific signaling pathway that might affect the maintenance of mouse submandibular gland (SMGs), RNA sequencing was performed. In SMGs, downregulated expression patterns of Rho-associated protein kinase (ROCK) signaling pathway-related genes, including Rhoa, Rhob, Rhoc, Rock1, and Rock2, were observed. Gene expression profiling analyses of these genes indicate that the ROCK signaling pathway is a potential signal for SMG maintenance.

Involvement of the Cyclic AMP-Protein Kinase A Pathway in Gap Junctional Communication in Preimplantation Mouse Embryos

  • Haengseok Song;Gye, Myung-Chan;Jun, Jin-Hyun
    • Animal cells and systems
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    • v.2 no.1
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    • pp.99-106
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    • 1998
  • In this study, we have examined the role of cAMP in gap junctional communication (GJC) in preimplantation mouse embryos. GJC was monitored by Lucifer Yellow (LY) injected into one blastomere of compacted embryos. The speed of GJC was defined as the time taken for the last blastomere of the embryo to become visibly fluorescent. The median time for 8-cell embrvos (140 sec) was similar to that for 16-cell (135 sec). To determine whether cAMP and cAMP-dependent protein kinase (PKA) are involved in the regulation of GJC, the effects of PKA inhibitor (H8) and cAMP analogues (Rp-cAMP and 8-Br-cAMP) on dye transfer between blastomeres of compacted embryos were examined. Some of the embryos treated with either H8 or Rp-cAMP failed to transfer LY to all blastomeres within 10 min. In contrast, 8-Br-cAMP speeded up fluorescent dye transfer. The median time to fill all blastomeres with LY was 140 sec in untreated controls and 90 sec in siblings treated with 8-Br-cAMP. Inhibition of PKA by H8 or Rp-cAMP induced delay or arrest in embryo development after compaction, but the increase of intracellular cAMP showed no effect. These findings suggest that GJC in preimplantation mouse embryos is regulated by cAMP-PKA pathway and transient interference by PKA inhibitors induces the developmental delay beyond compaction.

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TASK-2 Expression Levels are Increased in Mouse Cryopreserved Ovaries

  • Kang, Dawon;Choe, Changyong;Kim, Chang-Woon;Goo, Ae Jin;Han, Jaehee
    • Journal of Embryo Transfer
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    • v.30 no.4
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    • pp.277-282
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    • 2015
  • Cryopreservation affects osmotic tolerance and intracellular ion concentration through changes in expression levels of water and ion channels. Control of these changes is important for cell survival after cryopreservation. Relatively little is known about changes in $K^+$ channel expression compared to water channel expression. This study was performed to investigate changes in TASK-2 channel (KCNK5: potassium channel, subfamily K, member 5), a member of two-pore domain $K^+$ channel family, in cryopreserved mouse ovaries. Cryopreservation increased TASK-2 mRNA expression in mouse ovaries. In addition, TASK-2 protein expression was upregulated in vitrified and slowly frozen ovaries. TASK-2 protein was expressed in all area of granulosa cells that surround the oocyte within the follicle, except nucleus. Viability of cells overexpressed with TASK-2 was higher than that of vector-transfected cells. Our results found that TASK-2 expression was increased by cryopreservation and overexpression of TASK-2 decreased cryopreservation-induced cell death. These results suggest that TASK-2 upregulation might reduce cryodamage.

Cloning and Characterization of Bovine 5-Cytosine DNA Methyltransferase I cDNA

  • Lee, Poongyeon;Min, Kwan-Sik;Lee, Hyun-Gi;Kim, Soon-Jeung;Chung, Hee-Kyoung;Seo, Myung-Kyu;Lee, Yun-Keun;Kim, Sung-Woo;Park, Jin-Ki
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.39-39
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    • 2003
  • Methylation of DNA 5-cytosine in mammalian early embryo affects great deal in nuclear reprogramming and chromatin remodeling of developing embryo. Current efforts to clone and produce cloned animals including transgenic animals face various problems including low birth rate, irregular development, and so on. In this report, cDNA for the one of house keeping methyltransfcrase, Dnmt1 was cloned from bovine somatic tissues and was analyzed for its nucleotide sequences to investigate the structure and function of the gene in bovine early development. Nucleotide sequence of bovine Dnmt1 homologue showed 76.8% identity with that of human Dnmtl and 66.4% with mouse Dnmt1. Translated amino acid sequence showed 88.4% homology with human homologue and 75.8% homology with mouse counterpart. Three types of Dnmt1 are reported in mouse and human, and are likely present in bovine tissues. Understanding of role of Dnmt1 in bovine development may shed a light in the field of animal, especially bovine cloning.

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Inductional Expression of the Human Lactadherin Gene in Mouse Mammary Epithelial Cells

  • Kwon, Mo-Sun;Koo, Bon-Chul;Kim, Teoan
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.94-94
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    • 2002
  • Lactadherin (formerly known as BA46), a major glycoprotein of the human milk fat globule membrane, is abundant in human breast milk and breast carcinomas and may prevent symptomatic rotavirus infections. In this study, under the control of mouse whey acidic protein (WAP) promoter, the expression pattern of lactadherin (Ltd) in lactogenic hormone-dependent mouse mammary epithelial cell line HC11 were tested. pLNWLtd construct containing 2.4 kilobases of the WAP promoter and 1.5 kilobases of human lactadherin gene was stably transfered into HC11 cells using retroviral vector system. Integration and expression level of the transgene was estimated using PCR and RT-PCR, respectively. Prominent induction of Ltd gene under the WAS promoter was accomplished in the presence of insulin, hydrocortisone and prolactin, while induction with insulin alone resulted in lower expression. Our results demonstrate that the expression of the transgene is increased by synergistic effect of several lactogenic hormones, including insulin, hydrocortisone, and prolactin.

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The Effect of Electric Stimulation(anion pad) on the Maturation of Follicular Oocytes and the Cleavage of Fertilized Embryos of the Mouse (Electric Stimulation(음이온 pad)이 생쥐난자의 성숙 및 수정난의 난할에 미치는 영향)

  • Bae, In-Ha;Park, Won;Choi, Sung-Mi;Kim, Moon-Kyoo
    • Clinical and Experimental Reproductive Medicine
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    • v.23 no.3
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    • pp.293-301
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    • 1996
  • In the present study, mouse follicular oocytes and 2-cell embryos(late -zygote stage embryos included) were cultured on the electric pad for electric stimulation in the culture incubator. In addition, follicular oocytes and embryos were tested for maturation and development under higher temperature condition($39^{\circ}C$).Mouse follicular oocyte maturation were not affected by anion electric stimulation and there is no significant difference in GBVD and MI between the control and experiment group after 4hr culture. In the embryo culture, it was found that more morula and blastocyst were found in the electric stimulation group rather than the control(96hr). This may seem to be caused with cytoplasmic $Ca^{2+}$ transient rise by electric stimulation(anion pad). On the other hand higher temperature incubation ($39^{\circ}C$) on the anion pad caused all the embryos degenerated within $12h{\sim}24hr$ culture. This was quite different from large animal embryos(bovine, pig, sheep), in which beneficial effect of high temperature incubation for oocyte maturation and embryo development were found.

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Deformation prediction by a feed forward artificial neural network during mouse embryo micromanipulation

  • Abbasi, Ali A.;Vossoughi, G.R.;Ahmadian, M.T.
    • Animal cells and systems
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    • v.16 no.2
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    • pp.121-126
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    • 2012
  • In this study, a neural network (NN) modeling approach has been used to predict the mechanical and geometrical behaviors of mouse embryo cells. Two NN models have been implemented. In the first NN model dimple depth (w), dimple radius (a) and radius of the semi-circular curved surface of the cell (R) were used as inputs of the model while indentation force (f) was considered as output. In the second NN model, indentation force (f), dimple radius (a) and radius of the semi-circular curved surface of the cell (R) were considered as inputs of the model and dimple depth was predicted as the output of the model. In addition, sensitivity analysis has been carried out to investigate the influence of the significance of input parameters on the mechanical behavior of mouse embryos. Experimental data deduced by Fl$\ddot{u}$ckiger (2004) were collected to obtain training and test data for the NN. The results of these investigations show that the correlation values of the test and training data sets are between 0.9988 and 1.0000, and are in good agreement with the experimental observations.

Development of Mouse Embryos Reconstituted with 4-cell Nuclei at Metaphase with Nocodazole (Nocodazole로 분열기에 동조된 생쥐 4세포기 핵 유래 재구축란의 발생에 관한 연구)

  • 권오룡;하야우굉
    • Journal of Embryo Transfer
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    • v.11 no.2
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    • pp.95-101
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    • 1996
  • The present study investigated the effect of treatment with nocodazole on the efficiency of cell cycle synchronization and development of mouse 4-cefl embryos. In addition, developmental ability of reconstituted embryos that received nuclei from 4-cell embryos synchronized with nocodazole at M phase was examined in vitro and in vivo. (1) When 4-cell blastorneres exposed to culture medium containing l$\mu$g /ml nocodazole for 2, 4, 6 and 8 hrs, 40% (40/10l), 75% (l19/159), 85% (87/102) and 97% (155/160) of nuclei were synchronized at M phase, respectively. (2) Treated with nocodazole for 4 hrs, the proportion of 4-cell embryos developed to blastocysts (98%, 60/61) was not significanUy different from that of the control embryos (98%, 196/201). However, the developmental ability of 4-cell embryos treated for 8 (87%, P<0.05)and 12 hrs (76%, P

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