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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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Ultrarapid Freezing of Mouse 2-Cell Embryos (생쥐 2-세포기 수정란의 초급속동결)

  • 강만종;이철상;한용만;유대열;이경광
    • Korean Journal of Animal Reproduction
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    • v.14 no.1
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    • pp.9-16
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    • 1990
  • This study was carried out in order to investigate effects of cryoprotectant concentration and equilibration time on survival of ultrarapidly frozen 2-cell mouse embryos. Mouse 2-cell embryos, following dehydration by exposure to DMSO and sucrose, were directly immersed into liquid nitrogen and thawed in 37$^{\circ}C$ water. Viability was defined by development rate to the blastocyst stage after in vitro culture for 72 hours. The results are summarized as follows ; 1. When 0.25M of sucrose was added into the freezing medium at various concentrations of DMSO and dilution medium, higher development rate of embryo was obtained in 3.0M DMSO concentrations (82.6%). However, when sucrose concentraitons of 0.25 and 0.5 M were added to the freezing medium with 3.0 M DMSO and dilution medium, development rate of embryos were 81.7% and 24.1%, respectively. 2. In the equilibration time at room temperature, higher development rate was attained after short period of time (2.5min) in 3.0 M DMSO+0.25 M sucrose (85.9%). 3. The development rate of embryos at in vitro 2-cell, in vitro 2-cell, solution control and untreated control was 84.6%, 90.9%, 89.9%,, and 89.7%, respectively.

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Effect of capsaicin on murine lymphocyte functions and lymphoid tissue morphology

  • Lee, June-Chul;Park, Yeong-Min
    • IMMUNE NETWORK
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    • v.1 no.3
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    • pp.203-212
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    • 2001
  • Background: Rapid advances in neuroendocrine immunology have established the concept of bidirectional communication between the immune and neuroendocrine systems. Capsaicin suppresses the immune function by destroying substance P acting as mediatior of neuroendocrine immune system. Methods and Results: In this study, effect of capsaicin on mature murine lymphocyte functions and lymphoid tissue morphology was examined. Formally, capsaicin showed the strong cytotoxic effect on splenocyte over $10{\mu}g/ml$ concentration in citro. And proliferation and Th1-cytokine expression of splenic cells in mice that received high dose of capsaicin ($100{\mu}g/mouse$) were significantly diminished. However, low dose of capsaicin treatment did not influence these responses in vivo($1{\mu}g/mouse$) and in vitro (under $5{\mu}g/ml$). And the morphology of spleen and lymph nodes after capsaicin treatment was observed. In the spleen of mice injected with high dose of capsaicin (100, $200{\mu}g/mouse$), the size of white pulp was significantly decreased and the length of red pulp was increased, Moreover, vascularity index was diminished in a dose dependent manner. Conclusion: These results implies that immunosuppressive effect of capsaicin is associated with cytotoxic activity on lymphocyte, Th1-cytokine down-regulation and lymphoid tissue abnormalization, and this report is expected to give a hand to the study for the mechanism of action of neurotoxin of the immune system.

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Identification of a Functionally Relevant Signal Peptide of Mouse Ficolin A

  • Kwon, Sang-Hoon;Kim, Min-Soo;Kim, Dong-Bum;Lee, Keun-Wook;Choi, Soo-Young;Park, Jin-Seu;Kim, Yeon-Hyang;Lee, Young-Hee;Kwon, Hyung-Joo
    • BMB Reports
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    • v.40 no.4
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    • pp.532-538
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    • 2007
  • Mouse ficolin A is a plasma protein with lectin activity, and plays a role in host defense by binding carbohydrates, especially GlcNAc, on microorganisms. The ficolin A subunit consists of an N-terminal signal peptide, a collagen-like domain, and a C-terminal fibrinogen-like domain. In this study, we show that ficolin A can be synthesized and oligomerized in a cell and secreted into culture medium. We also identify a functionally relevant signal peptide of ficolin A by using MS/MS analysis to determine the N-terminal sequence of secreted ficolin A. When the signal peptide of mouse ficolin A was fused with enhanced green fluorescent protein (EGFP), EGFP was released into HEK 293 cell medium, suggesting that the signal peptide can efficiently direct ficolin A secretion. Moreover, our results suggest that the signal peptide of ficolin A has potential application for the production of useful secretory proteins.