• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2002년도 국제심포지엄
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• pp.115-115
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• 2002

To evaluate the correlations between the expression of cyclin B1 mRNA and protein after stimulation and oocyte activation and development of nuclear transferred mouse embryos, this study was performed. The oocyte activation was induced by 7% ethanol or 10$\mu\textrm{g}$/$m\ell$ Ca-ionophore without (single) or with (combined) 10$\mu\textrm{g}$/$m\ell$ cycloheximide (CH). Cyclin B1 mRNA and protein in mouse oocytes was evaluated by PCR and western blot. The activation and blastocyst development in both single (P<0.05) and combined (P<0.01) stimulation was higher than in non-activated group. The cyclin B1 mRNA and protein levels were significantly reduced in both single and combined stimulation groups (P<0.05), respectively. Cyclin B1 mRNA expression showed a negative correlation between activation and blastocyst development in both single and combined stimulation groups. And also the expression of cyclin B1 protein showed a negative correlation with between oocyte activation and blastocysts development in both single and combined stimulation groups. In conclusion, it may suggest that single and combined stimulation increases the oocyte activation and blastocyst development of nuclear transferred embryos, because it induces the degradation of cyclin B1 mRNA and protein after activation in enucleated mouse oocytes.

• Animal Bioscience
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• 제36권4호
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• pp.555-569
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• 2023

Objective: The objective of this study was to investigate the effects of N⁶-Methyladenosine modification-circRNA-zinc finger protein 638 (m⁶A-circRNA-ZNF638) on the induced activation of secondary hair follicle (SHF) stem cells with its potential mechanisms in cashmere goats. Methods: The m⁶A modification of ZNF638 was analyzed using methylation immunoprecipitation with real-time quantitative polymerase chain reaction technique in SHF stem cells. The effects of circRNA-ZNF638 on the induced activation of SHF stem cells in m⁶A dependence were evaluated through the overexpression of circRNA-ZNF638/its m⁶A-deficient mutants in circRNA-ZNF638 knockdown SHF stem cells. The competitive binding of miR-361-5p to circRNA-ZNF638/Wnt5a 3'- untranslated region was analyzed through Dual-luciferase reporter assay. Results: The m⁶A-circRNA-ZNF638 had significantly higher transcription at anagen SHF bulge of cashmere goats compared with that at telogen, as well as it positively regulated the induced activation of SHF-stem cells in cashmere goats. Mechanismly, m⁶A-circRNA-ZNF638 sponged miR-361-5p to heighten the transcriptional expression of Wnt5a gene in SHF-stem cells. We further demonstrated that the internal m⁶A modification within circRNA-ZNF638 is required for mediating the miR-361-5p/Wnt5a pathway to regulate the induced activation of SHF stem cells through an introducing of m⁶A-deficient mutant of circRNA-ZNF638. Conclusion: The circRNA-ZNF638 contributes the proper induced activation of SHF-stem cells in cashmere goats in m⁶A-dependent manner through miR-361-5p/Wnt5a axis.

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2002년도 국제심포지엄
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• pp.116-116
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• 2002

Further development of reconstructed embryos may be dependent upon the synchronization of donor nucleus and recipient cytoplasm at cell fusion, To control the synchronization of donor and recipient cells, the enucleated MII arrested oocytes are artificially stimulated prior to embryo reconstruction. Destruction of cyclin B results in the exit of cells from M-phase of cell cycle. This study was designed to investigate the effects of single or combined stimulation affected cyclin B1 mRNA and protein levels in mouse oocytes. The oocyte activation was induced by 7% ethanol or 10$\mu\textrm{g}$/$m\ell$ Ca-ionophore without (single) or with (combined) 10$\mu\textrm{g}$/$m\ell$ cycloheximide. Competitive quantitative PCR for cyclin Bl mRNA and western blot analysis for cyclin B1 protein was preformed in mouse oocytes. Cyclin B1 mRNA level was significantly reduced in single (P<0.05) and combined (P<0.05) stimulation groups. However, this level did not change in non-activated group and increased in intact group. Cyclin B1 protein level was also significantly reduced in both single (P<0.05) and combined (P<0.05) stimulation groups. In conclusion, single and combined stimulation induces the degradation of cyclin B1 mRNA and protein after activation in enucleated mouse oocytes.

• Molecules and Cells
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• 제42권5호
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• pp.426-439
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• 2019

Many small RNAs (sRNAs) regulate gene expression by base pairing to their target messenger RNAs (mRNAs) with the help of Hfq in Escherichia coli. The sRNA DsrA activates translation of the rpoS mRNA in an Hfq-dependent manner, but this activation ability was found to partially bypass Hfq when DsrA is overproduced. The precise mechanism by which DsrA bypasses Hfq is unknown. In this study, we constructed strains lacking all three rpoS-activating sRNAs (i.e., ArcZ, DsrA, and RprA) in $hfq^+$ and $Hfq^-$ backgrounds, and then artificially regulated the cellular DsrA concentration in these strains by controlling its ectopic expression. We then examined how the expression level of rpoS was altered by a change in the concentration of DsrA. We found that the translation and stability of the rpoS mRNA are both enhanced by physiological concentrations of DsrA regardless of Hfq, but that depletion of Hfq causes a rapid degradation of DsrA and thereby decreases rpoS mRNA stability. These results suggest that the observed Hfq dependency of DsrA-mediated rpoS activation mainly results from the destabilization of DsrA in the absence of Hfq, and that DsrA itself contributes to the translational activation and stability of the rpoS mRNA in an Hfq-independent manner.

• Genomics & Informatics
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• 제21권1호
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• pp.2.1-2.14
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• 2023

Microglia, similar to peripheral macrophages, are the primary immune cells of the central nervous system (CNS). Microglia exist in the resting state in the healthy CNS, but can be activated and polarized into either M1 or M2 subtypes for immune defense and the maintenance of CNS homeostasis by multiple stimuli. Several long noncoding RNAs (lncRNAs) mediate human inflammatory diseases and neuropathologies by regulating their target genes. However, the function of common lncRNAs that contribute to microglial activation remains unclear. Thus, we used bioinformatic approaches to identify common lncRNAs involved in microglial activation in vitro. Our study identified several lncRNAs as common regulators of microglial activation. We identified 283 common mRNAs and 53 common lncRNAs during mouse M1 microglial activation processes, whereas 26 common mRNAs and five common lncRNAs were identified during mouse M2 microglial activation processes. A total of 648 common mRNAs and 274 common lncRNAs were identified during the activation of human M1 microglia. In addition, we identified 1,920 common co-expressed pairs in mouse M1 activation processes and 25 common co-expressed pairs in mouse M2 activation processes. Our study provides a comprehensive understanding of common lncRNA expression profiles in microglial activation processes in vitro. The list of common lncRNAs identified in this study provides novel evidence and clues regarding the molecular mechanisms underlying microglial activation.

• 대한약리학회지
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• 제32권3호
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• pp.347-355
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• 1996

흰쥐 뇌하수체 전엽배양세포에서 PKC나 cAMP에 의한 LH 분비와 $LH{\beta}$subunit mRNA 증가과정에서 세포외 $Ca^{++}$의 역할을 검증하기 위하여 phorbol ester와 forskolin에 의해 촉진된 LH 분비와 $LH{\beta}$ subunit mRNA 수준에 미치는 EGTA와 verapamil의 영향을 조사하였다. Forskolin에 의해 촉진된 LH 분비와 $LH{\beta}$ subunit mRNA 수준은 $Ca^{++}$ chelator인 EGTA나 $Ca^{++}$ 채널차단제인 verapamil의 처리로 세포외부로부터 $Ca^{++}$의 이동을 억제시켰을때 모두 감소하였다. PMA에 의해 유도된 LH 분비는 EGTA와 verapamil 처리에 의해 영향을 받지 않았으나 PMA에 의해 촉진된 $LH{\beta}$ subunit mRNA는 억제되었다. 따라서 본 연구에 의하면 PKC 활성화나 세포내 cAMP 농도 증가는 $LH{\beta}$ subunit mRNA 수준을 증가시키며 이러한 과정은 세포외 $Ca^{++}$ 의존적인 것으로 생각된다. 그러나 PKC 활성화나 cAMP 증가에 의한 세포외 $Ca^{++}$의 역할이 서로 다른 양상을 보인다. PKC 활성화에 의한 LH 분비는 세포외 $Ca^{++}$에 비의존적이나 cAMP에 의해 촉진된 LH 분비는 세포외 $Ca^{++}$ 유입의 영향을 받는 것으로 사료된다.

• 생명과학회지
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• 제25권9호
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• pp.1043-1050
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• 2015

신경과흥분은 신경세포의 수지돌기 말단부에 있는 흥분성 수용체에 대한 과도한 자극에 의해서 신경세포가 손상을 받는 현상으로 transcription factor의 발현을 유도하여 통증을 유발하는 자극, 학습, 발작, 흥분, 신경변성, 저산소성 국소빈혈, 뇌신경손상, 신경절제, 약제내성 등의 원인이 된다. 신경과흥분은 정상농도 이상의 NMDA에 의해서도 유발되는데 본 논문에서는 mouse의 복강으로 과량의 NMDA를 투여하여 소뇌에서 RT-PCR 방법으로 Inducible transcription factors (Egr-1, c-jun, JunB, FosB) mRNAs의 상대적 발현량을 비교하였다. NMDA를 투여한 군에서 inducible transcription factors (Egr-1, C-Jun, JunB, FosB)가 투여량과 시간의 경과에 따라 다양한 발현의 변화를 보였으며, NMDA투여 후 일정한 시간에서 투여한 양에 대한 변화는 체중 g 당 5 μg의 NMDA투여한 경우에 현저한 변화가 나타났다. 조사한 transcription factor 중에서 JunB의 발현 변화가 다른 transcription factor보다 두드러지게 나타났다. NMDA 투여량이 일정할 때 투여 후 경과 시간에 따른 발현양상은 투여 후 24시간이 경과한 후에 발현의 변화가 두드러지게 증가하는 경향을 나타내었고 대부분 이 48시간 경과 후 발현이 최고치에 도달하였다. 이러한 결과는 과흥분이 유도된 소뇌에서의 유전자 발현의 변화를 2D-gel 또는 microarray와 같은 방법을 이용하여 세포 내의 전체 단백질 혹은 유전자의 변화를 관찰함으로써 NMDA 수용체의 과흥분에 의한 뇌세포의 사멸에 관련된 기전을 밝힐 수 있는 좋은 자료가 될 수 있을 것으로 기대된다.

• 한국임상수의학회지
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• 제34권3호
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• pp.172-177
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• 2017

Fucoidan, a cell wall polysaccharide found in the brown seaweed, is reported to have broad-spectrum biological activities. The objectives of this study were to examine the effect of fucoidan on prostaglandin $E_2$ ($PGE_2$) and cyclooxygenase-2 (COX-2) expression in lipopolysaccharide (LPS)-stimulated porcine peripheral blood mononuclear cells (PBMCs) and to determine whether these effects are involved in Akt activation. The levels of $PGE_2$ production in the culture supernatants from PBMCs were determined by the enzyme-linked immunosorbent assay (ELISA) kit and the levels of COX-2 mRNA were measured by real time polymerase chain reaction (RT-PCR). Akt activity was determined by Western blot analysis. Fucoidan in LPS-$na{\ddot{i}ve}$ PBMCs has no effect on $PGE_2$ production and COX-2 mRNA expression. Furthermore, fucoidan does not affect Akt activation in LPS- $na{\ddot{i}ve}$ PBMCs. However, $PGE_2$ production and COX-2 mRNA expression on PBMCs were remarkably enhanced by LPS stimulation. Akt activity was also increased by LPS. Increasing effects of $PGE_2$ production and COX-2 mRNA expression in PBMCs induced by LPS were suppressed by addition of fucoidan. In addition, fucoidan reduced an increase in Akt activity in LPS-stimulated PBMCs. These results suggested that fucoidan exerts potent anti-inflammatory properties by suppression of $PGE_2$ production, COX-2 mRNA expression and Akt activation in LPS-stimulated PBMCs.

• 한방안이비인후피부과학회지
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• 제21권2호
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• pp.46-53
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• 2008

Background and Objectives : The aim of this study was to investigate the anti-oxidant, anti-allergic and anti-inflammatory ability of the Taglisodog-eum(SHE) extract on the RAW 264.7 and EL4 cells Materials and Methods : Three types of experiments were implemented for this study: first, the experiment to study the anti-oxidant effect of SHE using Riboflavin; second, in vitro experiment to investigate the inhibition of Th 2 cell differentiation by SHE using EL4 cells (IL-4 mRNA expression); third, the suppression of $NF-{\kappa}B$ activation using RAW 264.7 cells (iNOS and COX-2 mRNA expression). Results : The anti-oxidant ability of SHE were dose-dependantly increased. From in vitro, the LPS-induced iNOS and COX-2 mRNA expression were dose-dependantly decreased in the RAW264.7 cells treated with SHE and the PMA-induced IL-4 mRNA expression were also dose-dependantly decreased in EL4 cells. $NF-{\kappa}B$ activation was suppressed, and iNOS & COX-2 production were inhibited by SHE Conclusion : The results suggest that SHE has dose-dependant anti-oxidant ability, and has anti-allergic and anti-inflammatory effects through the suppression of $NF-{\kappa}B$ activation and the inhibition of Th 2 cell differentiation.

• 동의신경정신과학회지
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• 제22권1호
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• pp.37-51
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• 2011

Objectives : This experiment was designed to investigate the effect of Gamisoyo-san(Jiaweixiaoyaosan, SYS) on serotonin activity of P815 Mast Cell. Methods : The effects of SYS on activation of TPH-1 mRNA and AAADC mRNA in P815 mast cell were investigated. The effect of SYS on content of serotonin in P815 mast cell was investigated. The effects of SYS on activation of DPPH and SOD in P815 mast cell were investigated. Results : 1. The SYS increased the activation of SOD and DPPH in P815 mast cell. 2. The SYS decreased the manifestation TPH-1 mRNA in P815 mast cell. 3. The SYS decreased the manifestation AAADC mRNA in P815 mast cell. Conclusions : This experiment shows that Gamisoyo-san might not be effective for treating depression in terms of biogenic amine theory. However, Gamisoyo-san showed significant anti-oxidative effect and it can not yet be ruled out for treating depression. Therefore, pathogenesis of depression and clinical research of Gamisoyo-san is suggested for future research.