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Obtainment and Characterization of Brain Tumor Cell Using Vasopressin-SV40 T Ag Transgenic Mouse
Kim Sung-Hyun,Lee Eun-Ju,Kim Myoung-Li,Park Jun-Hong,Cho Kyoungin,Jung Boo-Kyung,Kim Hee-Chul,Hwnag Sol-Ha,Lee Hoon-Taek,Ryoo Zae-Young 한국발생생물학회 2003 한국발생생물학회 학술발표대회 Vol.2003 No.1
In previous reports, pVPSV.IGR2.1 transgenic mouse were described that brain tumor and lymphoma by reason of Vasopressin-SV40 T antigen. In this study, we produced pVPSV.IGR3.6 transgenic mouse that used pVPSV.IGR3.6 vector. Expression of transgene was vary different in transgenic mouse. We obtained 6 transgenic mouse line, moreover they had died at the age of 2-6 weeks without transmitting the transgene to their offspring, and had tumorigenesis on same location with pVPSV.IGR2.1 transgenic mouse. Only a founder mouse was investigated for expression of fusion gene. Here we extended this transgenic approach to the study of tumor progression. From the mouse, we confirmed brain tumor cell, after then cultured for investigate characterization. In this report, we demonstrate that reduction of survival rate in transgenic mouse fused vasopressin gene length, acquisition of brain tumor cell, composition with astrocyte cells and neuronal cells. Finally, cells had no change with increase of passage.
Kim Sung-Hyun,Lee Eun-Ju,Kim Myoung-Li,Park Jun-Hong,Cho Kyoungin,Jung Boo-Kyung,Kim Hee-Chul,Hwnag Sol-Ha,Lee Hoon-Taek,Ryoo Zae-Young 한국발생생물학회 2003 한국발생생물학회 학술발표대회 Vol.2003 No.1
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 /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 ) 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.
Kim, Jinmahn,Yeon, Jihye,Choi, Seong-Kyoon,Huh, Yang Hoon,Fang, Zi,Park, Seo Jin,Kim, Myoung Ok,Ryoo, Zae Young,Kang, Kyeongjin,Kweon, Hee-Seok,Jeon, Won Bae,Li, Chris,Kim, Kyuhyung Public Library of Science 2015 PLoS genetics Vol.11 No.8
<▼1><P>The expression of specific transcription factors determines the differentiated features of postmitotic neurons. However, the mechanism by which specific molecules determine neuronal cell fate and the extent to which the functions of transcription factors are conserved in evolution are not fully understood. In <I>C</I>. <I>elegans</I>, the cholinergic and peptidergic SMB sensory/inter/motor neurons innervate muscle quadrants in the head and control the amplitude of sinusoidal movement. Here we show that the LIM homeobox protein LIM-4 determines neuronal characteristics of the SMB neurons. In <I>lim-4</I> mutant animals, expression of terminal differentiation genes, such as the cholinergic gene battery and the <I>flp-12</I> neuropeptide gene, is completely abolished and thus the function of the SMB neurons is compromised. LIM-4 activity promotes SMB identity by directly regulating the expression of the SMB marker genes via a distinct <I>cis</I>-regulatory motif. Two human LIM-4 orthologs, LHX6 and LHX8, functionally substitute for LIM-4 in <I>C</I>. <I>elegans</I>. Furthermore, <I>C</I>. <I>elegans</I> LIM-4 or human LHX6 can induce cholinergic and peptidergic characteristics in the human neuronal cell lines. Our results indicate that the evolutionarily conserved LIM-4/LHX6 homeodomain proteins function in generation of precise neuronal subtypes.</P></▼1><▼2><P><B>Author Summary</B></P><P>The correct generation and maintenance of the nervous system is critical for the animal’s life. Dysregulation of these processes leads to multiple neurodevelopmental disorders. It has been a daunting challenge not only to identify the developmental mechanisms that determine neuronal cell fate, but also to understand the extent to which the mechanisms are evolutionarily conserved. Here, we describe a developmental mechanism that determines the fate of a specific cholinergic and peptidergic neuronal type in <I>C</I>. <I>elegans</I>. We show that the <I>lim-4</I> LIM homeodomain transcription factor is necessary and sufficient to promote and maintain the specific cholinergic and peptidergic properties and functions via binding to unique DNA sequences. We also demonstrate that <I>C</I>. <I>elegans lim-4</I> and human <I>LHX6</I> show striking functional similarity; specifically, <I>C</I>. <I>elegans</I> LIM-4 or human LHX6 can induce cholinergic and peptidergic characteristics in human neuronal cell lines. Given the high conservation of these transcription factors, these developmental mechanisms are likely to be generally applicable in the nervous system of other organisms as well.</P></▼2>
Li, Yan,Kim, Ryunhee,Cho, Yi Sul,Song, Woo Seok,Kim, Doyoun,Kim, Kyungdeok,Roh, Junyeop Daniel,Chung, Changuk,Park, Hanwool,Yang, Esther,Kim, Soo-Jeong,Ko, Jaewon,Kim, Hyun,Kim, Myoung-Hwan,Bae, Yong- Society for Neuroscience 2018 The Journal of neuroscience Vol.38 No.26
<P>SALM1 (SALM (synaptic adhesion-like molecule), also known as LRFN2 (leucine rich repeat and fibronectin type III domain containing), is a postsynaptic density (PSD)-95-interacting synaptic adhesion molecule implicated in the regulation of NMDA receptor (NMDAR) clustering largely based on in vitro data, although its in vivo functions remain unclear. Here, we found that mice lacking SALM1/LRFN2 (Lrfn2(-/-) mice) show a normal density of excitatory synapses but altered excitatory synaptic function, including enhanced NMDAR-dependent synaptic transmission but suppressed NMDAR-dependent synaptic plasticity in the hippocampal CA1 region. Unexpectedly, SALM1expression was detected in both glutamatergic and GABAergic neurons and Lrfn2(-/-) CA1 pyramidal neurons showed decreases in the density of inhibitory synapses and the frequency of spontaneous inhibitory synaptic transmission. Behaviorally, ultrasonic vocalization was suppressed in Lrfn2(-/-) pups separated from their mothers and acoustic startle was enhanced, but locomotion, anxiety-like behavior, social interaction, repetitive behaviors, and learning and memory were largely normal in adult male Lrfn2(-/-) mice. These results suggest that SALM1/LRFN2 regulates excitatory synapse function, inhibitory synapse development, and social communication and startle behaviors in mice.</P>
랫드의 간에서 다양한 농도의 아플라톡신 투여에 의한 DNA Adduct의 형성과 Ras의 발현양상
김태명(Tae Myoung Kim),허진주(Jin Joo Hue),리란(Lan Li),김대중(Dae Joong Kim),남상윤(Sang Yoon Nam),윤영원(Young Won Yun),이범준(Beom Jun Lee) 한국독성학회 2005 Toxicological Research Vol.21 No.4
Aflatoxins are produced by Aspergillus flavus, parasiticus that grows in improperly stored cereals. Aflatoxin B₁ (AFB₁) is a potent hepatocarcinogen in a variety of experimental animals including human beings. In spite of a high attention to the hepatocarcinogenecity of aflatoxins, the relative toxicity of other types (AFB₂, AFG₁ and AFG₂) of the toxins is not fully clarified. Sprague-Dawley male rats were orally administered with AFB₁, AFB₂, AFG₁ and AFG₂ at the dose of 250, 1250, and 2500 μg/kg body weight. Animals were then killed at 12, 24 or 48 hrs following aflatoxin adminstration. Subsequently the relative weight of liver was measured and histopathological examination on the liver was performed. Level of 8-OxodG and expression of ras gene in the liver was determined. The relative liver weights at high doses of AFB₁ and AFG₁ was significantly low. The treatment of AFB₁ at the high dose of 2500 ㎍/㎏ showed vacuolar degeneration and centrilobular hepatic necrosis with inflammatory cells. The pathological changes by AFB₂, AFG₁ and AFG₂ were not clearly found. The formation of 8-OxodG by AFB₁ increased in a dose-dependent manner up to 24 hrs after a single treatment of AFB₁ thereafter decreased to the level of the control. The treatments of AFB₂, AFG₁ and AFG₂ showed an inconsistent pattern in the formation of 8-OxodG in the liver of rats with increasing time. The expression of ras oncogene in the liver by AFB₁ at the dose of 1250 ㎍/㎏ was increased twice compared to the control. The treatments of AFB₂, AFG₁ and AFG₂ at all doses decreased the expression of ras in the liver. These results in the present study indicate that AFB₁ among aflatoxins with low comparable levels is the most toxic as determined by early biomarkers such as 8-OxodG formation and ras expression. However, the levels of 8-OxodG and ras as biomarkers were not useful to predict the relative hepatocarcinogenicity of aflatoxins to AFB₁ in the present model. Further studies are required to look for other biomarkers to predict carcinogenic potency of aflatoxins.
Lee, Hyun Gyu,Cho, Nam-chul,Jeong, Ae Jin,Li, Yu-Chen,Rhie, Sung-Ja,Choi, Jung Sook,Lee, Kwang-Ho,Kim, Youngsoo,Kim, Yong-Nyun,Kim, Myoung-Hwan,Pae, Ae Nim,Ye, Sang-Kyu,Kim, Byung-Hak Elsevier 2016 The Journal of investigative dermatology Vol.136 No.1
<P>T-cell-mediated immune responses play an important role in body protection. However, aberrantly activated immune responses are responsible for inflammatory and autoimmune diseases. The regulation of pathologic immune responses may be a potential therapeutic strategy for the treatment of these diseases. Despite that multiple pharmacologic properties of benzoxathiole derivatives have been defined, the molecular mechanisms underlying these properties remain to be clarified. Here, we demonstrated the benzoxathiole derivative 2-cyclohexylimino-6-methyl-6,7-dihydro-5H-benzo[1,3]oxathiol-4-one (BOT-4-one) regulated immune responses and ameliorated experimentally induced inflammatory skin diseases both in vitro and in vivo. BOT-4-one inhibited the differentiation of CD4(+) T-cell subsets by regulating the expression and production of T-cell lineage-specific master transcription factors and cytokines and activating the signal transducer and activator of transcription proteins. In addition, BOT-4-one inhibited TCR-mediated Akt and NF-kappa B signaling. Topical application of BOT-4-one ameliorated experimentally induced inflammatory skin diseases in mice models such as 2,4,6-trinitrochlorobenzene-induced contact and atopic dermatitis and IL-23-induced psoriasis-like skin inflammation. Our study demonstrated that BOT-4-one ameliorates inflammatory skin diseases by suppressing the pathogenic CD4(+) T cell differentiation and overall immune responses.</P>
Kim, Myoung-Hee,Woo, Hee-Gweon,Park, Woo Jin,Sohn, Honglae,Chu, Bryan Tsu-Te,Li, Hong,Ko, Young Chun American Scientific Publishers 2010 Journal of Nanoscience and Nanotechnology Vol.10 No.5
<P>The co-dehydrocoupling at ambient air atmosphere of 1,1-dihydrotetraphenylsilole 1 and 1,1-dihydrotetraphenylgermole 2 (9:1 mole ratio) with 2 mol% of AgNO3 in toluene at 90 degrees C produces optoelectronic poly(silole-co-germole)s 3 in high yield. The copolymer mainly has Si-Si bonds (and Si-Ge/Ge-Ge bonds in minor) along with the small portion of Si-O/Ge-O bonds in the polymer backbone chain. While Ag2SO4 is also a good catalyst as AgNO3 for the co-dehydrocoupling, AgI is a moderate catalyst. However, CP2Co, Cp2Ni, Cp2ZrCl2/Red-Al, AgX (X = F, Cl, Br) and AgI@MWCNT do not show appreciable catalytic activity. The silver complexes (AgNO3, Ag2SO4, AgI) transformed to colloidal silver nanoparticles during the catalytic reaction. The co-dehydrocoupling of 1 and 2 with AgNO3 even at dry nitrogen atmosphere is occurred, supporting that the oxidation of NO3- ion to NO2 is only the possible oxygen source, but not from the adventitious moisture in air. All cyclodextrins (sigma, beta, and gamma forms) considerably deteriorated the co-dehydrocoupling of 1 and 2 probably by forming toluene-insoluble inclusion complexes and by encapsulating SiH2 moiety. The resulting copolymer emits green light at 521 nm and is electroluminescent at 523 nm, which is similar to the polysilole.</P>