도파민 수용체 제 2 형 Exon 8 유전자 다형성과 알코올 의존의 임상 양상

이소희(So Hee Lee),이분희(Bun-Hee Lee),이준석(Jun-Seok Lee),채영규(Young Gyu Chai),최미란(Mi Ran Choi),한달무리(Dal Mu Ri Han),지 홍(Hong Ji),장경호(Gyeong-Ho Jang),신혜은(Hye Eun Shin),최인근(Ihn Geun Choi) 한국중독정신의학회 2013 중독정신의학 Vol.17 No.1

Objectives : Various single nucleotide polymorphisms (SNPs) of dopamine D2 receptor (DRD2) genes have been reported to be involved in the susceptibility to alcohol dependence. We investigated the association of exon 8 (E8) SNP in the DRD2 gene with alcohol dependence in Korean subjects and examined their associations with severities of depression, craving, and alcohol-withdrawal symptoms. Methods : The DRD2 E8 A/G polymorphisms were genotyped in a case-control sample consisting of 245 alcohol-dependent (AD) patients and 130 healthy controls (HC). Severities of de-pression, craving, and alcohol-withdrawal symptoms were examined with BDI, PACS, and CIWA-Ar in AD patients. Results : The genotype or allele frequencies of DRD2 E8 A/G had no significant differences between AD and HC groups (χ 2 = 2.659, p=0.265; χ 2 =2.694, p=0.107). AD with A/A genotype and A allele in E8 had significantly higher BDI scores (F=3.705, p=0.029; t=2.073, p=0.040), although there were no significant differences in PACS and CIWA-Ar scores among genotypes or alleles of E8 A/G. Conclusion : The present study suggests the possibility that DRD2 E8 polymorphisms have an impact on the AD phenotype, such as an increased depressiveness in Korean patients with alcohol dependence.

에탄올 처리에 의한 흰쥐 신경아교종(Glioma) 세포에서의 유전자 발현 - DNA 칩을 이용한 분석 -

이소희,오동열,한진희,최인근,전양환,이준노,이태경,정종현,정경화,채영규,Lee, So Hee,Oh, Dong-Yul,Han, Jin-Hee,Choi, Ihn-Geun,Jeon, Yang-Whan,Lee, Joon-Noh,Lee, Tae Kyung,Jeong, Jong-Hyun,Jung, Kyung Hwa,Chai, Young-Gyu 대한생물정신의학회 2007 생물정신의학 Vol.14 No.2

연구목적: 알코올의존에 내재된 분자생물학적 기전을 이해하고 알코올리즘 치료 약물의 새로운 표적을 알아내기 위해서는, 알코올에 반응하는 유전자 혹은 반응 경로를 알아내는 것이 필요하다. DNA microarray 기법의 발달로 고전적 연구 방법과 달리 동시에 수천 수만개의 유전자의 표현을 검사하는 것이 가능하게 되었다. 본 연구에서는 알코올을 흰쥐의 신경아교종 세포에 처리했을 때 어떤 유전자의 발현을 조절하는지 DNA microarray를 이용하여 알아보고자 하였다. 방 법: 흰쥐 신경아교종 C6 세포주를 배양하여 에탄올 처리하고 총 RNA를 분리한 후 유전자 발현 양상을 조사하기 위해 cDNA microarray를 수행하였다. 결 과: 에탄올 처리군과 대조군간의 유전자 발현의 차이를 비교 분석한 결과 에탄올이 처리된 군에서 대조군에 비해 15개의 유전자가 발현이 증가하였고 12개의 유전자가 발현이 감소하였다. 발현이 증가한 유전자는 Orthodenticle(Drosophila) homolog 1, procollagen type II, adenosine A2a receptor, GATA-bindning protein2를 포함하고 있었고, 발현이 감소한 유전자는 diacylglycerol kinase beta, PRKC, Protein phosphatase 1, clathrin-associated protein 17, nucleoporin p58, proteasome를 포함하였다. 결 론: 흰쥐의 신경아교종 세포주에 알코올을 처치하였을 때 급성기에 알코올에 반응하여 발현이 증가하거나 감소한 유전자는 전반적으로 전사의 조절, 신호전달체계, 허혈성 뇌손상의 중재, 신경세포의 퇴행에 관여하는 것들이었다. 본 연구는 유전자 발현 시스템을 이용하여 에탄올에 반응하는 새로운 후보 유전자들을 관찰하였다는데 의의가 있다. Objetives : Identification of target genes for ethanol in neurons is important for understanding its molecular and cellular mechanism of action and the neuropathological changes seen in alcoholics. The purpose of this study is to identify of altered gene expression after acute treatmet of ethanol in rat gliom cells. Methods : We used high density cDNA microarray chip to measure the expression patterns of multiple genes in cultured rat glioma cells. DNA microarrays allow for the simultaneous measurement of the expression of several hundreds of genes. Results : After comparing hybridized signals between control and ethanol treated groups, we found that treatment with ethanol increased the expression of 15 genes and decreased the expression of 12 genes. Upregulated genes included Orthodenticle(Drosophila) homolog 1, procollagen type II, adenosine A2a receptor, GATA bindning protein 2. Downregulated genes included diacylglycerol kinase beta, PRKC, Protein phosphatase 1, clathrin-associated protein 17, nucleoporin p58, proteasome. Conclusion : The gene changes noted were those related to the regulation of transcription, signal transduction, second messenger systems. modulation of ischemic brain injury, and neurodengeneration. Although some of the genes were previously known to be ethanol responsive, we have for the most part identified novel genes involved in the brain response to ethanol.

생쥐 초기배아의 유전자 활성에 미치는 Protein Kinase Inhibitors의 영향

이정은,채영규,배인하,윤용달,김문규,Lee, Jeong-Eun,Chai, Young-Gyu,Bae, In-Ha,Yoon, Young-Dal,Kim, Moon-Kyoo 대한생식의학회 1995 Clinical and Experimental Reproductive Medicine Vol.22 No.2

Transcriptional activation of the embryonic genome initiates at 2-cell stage in mouse embryo and is characterized by the synthesis of TRC which is restricted to 2-cell stage. To investigate the roles of various protein kinases on the embryonic gene activation, the effects of protein kinase inhibitors on in vitro development and protein synthetic profiles of the early mouse embryos were examinded. None of ${\alpna}-amanitin$ which is a mRNA synthetic inhibitor, H8 which is a PKA inhibitor, and H7 which is a PKC inhibitor, affected on first cleavage of mouse 1-cell embryos in vitro. However, all of these drugs inhibited the second cleavage. When the drugs were removed following treatment for 6 hours, H8 or H7 treatment showed little inhibition on subsequent development of 1-cell embryos to 2-cell stage or further. In contrast, ${\alpna}-amanitin$ irreversibly inhibited the development of 1-cell embryos to 2-cell stage following removal of the drug. Genistein, a TPK inhibitor, inhibited both the first cleavage of 1-cell embryos and the second cleavage of 2-cell embryos, suggesting that TPK activity may be important during the early cleavages. All of the above four drugs inhibited TRC synthesis as shown by the fluorographic analysis of $[^{35}S]-Met$ labeled protein profiles. When late 1-cell embryos were treated with H7 and analyzed synthetic patterns of $[^{35}S]-Met$ labeled protein, the quantitative differences of protein synthesis on SDS-PAGE appeared on 77 kD and 33 kD region at $32{\sim}38$ hours post hCG. From these studies, transcriptional activation of embryonic genome is not essenting to the mouse 1-cell embryos to develop to 2-cell stage. Hawever, TPK activity is reguisite for both the first cleavage and second cleavage. Similarly, both PKC and PKA activities are required for the second cleavage of mouse embryos, but not for the first cleavage.

카드뮴 저항성인 Chlamydomonas reinhardtii 의 부분 특성

이창균,채영규,최영길 ( C K Lee,Y G Chai,Y K Choi ) 한국하천호수학회 1991 생태와 환경 Vol.24 No.2

The effects of heavy metals on the growth and pigment content of green algae were examined. For heavy metals toxicity, C. reinhardtii 21 gr^+, cell wall mutant CW-15^- and Cd-tolerant population CW-15^r were used. These strains were exposed to the various concentration of CdCl₂ and CuSO₄, in a Tris-acetate phosphate medium. A CW -15^r which derived from a Cd-sensitive cell wall mutant CW-15^- shows that CdCl₂ toxicity was 7 times higher than that of CW-15^-. The 50% inhibition of cell growth and chlorophyll content for Chlamydomonas were found at Cd concentrations of about 60 μM, 400 μM for CW-15^-, CW-15^r and at Cu concentrations of 100 μM, 80 μM for 21 gr^+, CW-15^-, respectively. Results suggested that cell wall mutant Chlamydomonas is adjusted to survive in their harsh heavy metal environment.

재조함 인성장호르몬의 in vitro 풀림과 재접힘 과정의 구조변화 모니터링

조태훈,채영규,안상점,이은규,Cho, Tae-Hoon,Chai, Young-Kyu,Ahn, Sang-Jeom,Lee, Eun-Kyu 한국생물공학회 1999 KSBB Journal Vol.14 No.6

재조합 인성장호르몬을 사용하여 in vitro 재접힘 공정(풀림, 희석에 의한 공기 중 산화, 그리고 투석)을 수행하였다. 표면소수성이 풀림-재접힘 공정에 중요한 역할을 한다는 것을 형광값의 변화를 통하여 알 수 있었다. 변성제의 intermediate 농도는 Urea와 Gu-HCl 경우 하나의 peak로 SDS와 Sarkosyl의 경우 두개의 peak로 나타났다. 형광값의 변화 중 특이한 점은 Urea의 경우 공기 중 산화와 투석 중의 후반부에 형광값이 증가한다는 것이다. 따라서 공기 중 산화도중 형광값이 증가하기 전에 투석을 시킨 결과 형광값이 증가를 막을 수 있었다. 아직 이 원인에 대해 자세히 알 수 없지만 계속 실험 중에 있다. 이번 실험에서 표면소수성 변화와 연관시켜 fluorescence를 이황화결합에 의한 산화된 형태를 알아보기 위한 방법으로 RP-HPLC를 마지막으로 단백질의 2차원적인 구조를 알아 보기 위해 CD를 사용하였다. CD측정 결과 Gu-HCl보다 SDS의 경우 ${\alpha}$-helices의 파괴가 더 많음을 볼 수 있었다. 재접힘된 rhGH는 본래의 2차원적 구조의 90%이상을 얻을 수 있었다. 이 실험이 기지는 의의는 이 모든 실험결과를 토대로 단백질 재접힘을 모니터링 하였다는 점이다. 즉, 형광값의 변화를 통하여 형광값이 증가하는 것은 표면 소수성이 증가함을 보이는 것으로 단백질의 풀림이 일어난 것이고 3차원적 구조가 깨지고 2차원 구조를 알아 볼 수 있는 ${\alpha}$-helices의 감소를 의미하였다. 이와는 반대로 형광값이 감소하는 것을 통해 재접힘이 일어남을 알 수 있었고, 이러한 결과를 바탕으로 단백질의 재접힘 공정의 변화과정을 형광값을 통하여 모니터링 할 수 있었다. 또한 이 실험의 목적 단백질은 rhGH이지만 다른 단백질에 적용이 될 경우 단백질 재접힘 과정을 수시로 모니터링하고 상태를 예측할 수 있으므로 산업현장에서 소량의 sample로 재접힘 상태를 쉽고 빠르게 판단할 수 있을 것이다. 단백질 재접힘 과정에서 이러한 개념의 성공적 도입은 단백질 회수 수율을 높임으로써 생물분리공정 분야의 기술 발전에 이바지 하리라 사료된다. Using recombinant human growth hormone as a model protein, we carried out unfolding by adding a denaturant such as urea, guanidine HCl, or SDS followed by refolding by dilution and dialysis. The objectives were to monitor the structural changes during in vitro refolding process and, based on the results, to develop a quantitative method of refolding progress assessment. The changes in surface hydrophobicity were measured by fluorescence tagging of 1-anilinonaphthalene-8-sulfonate(1,8-ANS) to the hydrophobic portions, and those in the secondary structure were monitored by using far UV-CD(circular dichroism) spectroscopy. Also, we used RP-HPLC to separate and quantify the folded and unfolded proteins to correlate the result with the structure analysis. Our results indicate the surface hydrophobicity are well correlated with the formations of the secondary structure, primarily ${\alpha}$-helices, as well as the disulfide bridges. We expect this monitoring technique can be applied in industrial fields as a means to quantitatively assess the progress of in-vitro refolding of recombinant proteins.

알코올의존의 Lesch의 유형학에 따른 신경생물학적 특성 및 임상적 특성

최미란,채영규,이분희,이준석,Choi, Mi-Ran,Choi, Young-Gyu,Lee, Bun-Hee,Lee, Jun-Seok 대한생물정신의학회 2011 생물정신의학 Vol.18 No.3

Objectives Many studies have suggested different neurobiological findings and clinical courses in alcoholism. Recently, subtyping in alcohol dependence has become essential to overcome the heterogeneity of patients. Among several criteria of subtypes, Lesch's typology is proposed to integrate biological, social, and psychological factors. This review provides neurobiological findings and treatment-responses of alcohol dependence according to Lesch's typology. Method We searched the international published medical literature using the search terms 'Lesch's typology' and 'alcohol dependence' and using the limits 'human'. Results We identified 17 studies with subjects of alcohol dependence according to Lesch's typology. Conclusion They indicated that each subtype of Lesch's typology can have specific neurobiological factors and different clinical responses as follows. Lesch's subtype 1 is characterized by severe withdrawal symptoms and associated with elevated glutamate and homocysteine. Lesch's subtype 2 is defined by individuals who drink alcohol as self-medication for anxiety. Their craving has significant positive correlations with prolactin, leptin level, or intake-volume (vasopressin). Lesch's subtype 4 is related to cerebral dysfunction and associated with increased glutamate and left-handedness. Clinical trials showed that naltrexone was effective in Lesch's subtype 3 and 4 patients, while acamprosate was effective in the subtypes 1 and 2.

장기간 플루세틴 처리에 의한 흰쥐 해마에서의 NCAM140 유전자 발현의 증가

최미란,채영규,정경화,백승연,김석현,노성원,최준호,이준석,최인근,양병환,Choi, Mi Ran,Chai, Young Gyu,Jung, Kyoung Hwa,Baik, Seung Youn,Kim, Seok Hyeon,Roh, Sungwon,Choi, Joonho,Lee, Jun-Seok,Choi, Ihn Geun,Yang, Byung-Hwan 대한생물정신의학회 2009 생물정신의학 Vol.16 No.1

Objectives : Most of the mechanisms reported for antidepressant drugs are the enhancement of neurite outgrowth and neuronal survival in the rat hippocampus. Neural cell adhesion molecule 140(NCAM140) has been implicated as having a role in cell-cell adhesion, neurite outgrowth, and synaptic plasticity. In this report, we have performed to elucidate a correlation among chronic antidepressant treatments, NCAM140 expression, and activation of phosphorylated cyclicAMP responsive element binding protein(pCREB) which is a downstream molecule of NCAM140-mediated intracellular signaling pathway in the rat hippocampus. Methods : Fluoxetine(10mg/kg) was injected acutely(daily injection for 5days) or chronically(daily injection for 14days) in adult rats. RNA and protein were extracted from the rat hippocampus, respectively. Real-time RT-PCR was performed to analyze the expression pattern of NCAM140 gene and western blot analyses for the activation of the phosphorylation ratio of CREB. Results : Chronic fluoxetine treatments increased NCAM140 expression 1.3 times higher than control in rat hippocampus. pCREB immunoreactivity in the rat hippocampus with chronic fluoxetine treatment was increased 4.0 times higher than that of control. Conclusion : Chronic fluoxetine treatment increased NCAM140 expression and pCREB activity in the rat hippocampus. Our data suggest that NCAM140 and pCREB may play a role in the clinical efficacy of antidepressants promoting the neurite outgrowth and neuronal survival.

Chlamydomonas reinhardtii Glutamine Synthetase 에 대한 Phosphinothicin 의억제

이창균,정준희,채영규,최영길 ( C K Lee,J H Jung,Y G Chai,Y K Choi ) 한국하천호수학회 1992 생태와 환경 Vol.25 No.2

Kinetics for the inhibition of glutamine synthetase from Chlamydomonas reinhardtii by phosphinothricin (PPT) have been examined. Inhibition by PPT was competitive, and apparent Ki value of 7.0μM was determined. This value of Ki was similar to the other types of glutamine synthetase studied from the Escherichia coli and Asparagus sprengeri.

한국인 알코올리즘과 Catechol-O-methyltransferase(COMT) 유전자 다형성의 연합

김민정,양병환,이정식,채영규,박택규,Kim, Min-Jung,Yang, Byung-Hwan,Lee, Jung-Sik,Chai, Young-Gyu,Park, Taek-Kyu 대한생물정신의학회 2001 생물정신의학 Vol.8 No.1

An association study with Korean alcoholic patients(n=50) and normal controls(n=53) was performed to find the relationship between catechol-O-methyltransferase(COMT) gene polymorphism and alcoholism using polymerase chain reaction-restriction fragment length polymorphism. When we compared the allele and genotype frequencies of Nla III COMT gene polymorphism in alcoholism and normal controls, there was no significant difference between two groups. Our results do not support an association between the Nla III polymorphism of COMT gene and alcoholism.

Chlamydomonas reinhardtii 의 생장과 색소함량에 미치는 제초제의 영향

이창균,정준희,최영길,한명수,채영규 ( C K Lee,J H Chung,Y K Choi,M S Han,Y G Chai ) 한국하천호수학회 1990 생태와 환경 Vol.23 No.3

The effects of herbicides on the growth and pigment content of green algae were examined. Chlamydomonas reinhardtii was exposed to herbicides (glyphosate, phosphinothricin, simazine, alachlor and 2,4-D) concentration ranging from 1.0 x 10^(-6)M to 1.0 x 10^(-2) M contained in a high salt minimal medium. With increasing concentration of herbicides, a significant decrease in the cell number, and pigment content was observed. EC_(50), the concentration at which growth rates were reduced to a half, were 3.73 x 10^(-5) M, 3.18 x 10^(-5) M, 6.53 x 10^(-5) M, 4.75 x 10^(-7) M and 1.05 x 10^(-2) M for glyphosate, phosphinothricin, simazine, alachlor and 2,4-D, respectively.