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RISS 인기검색어
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- 저자 : Sung-Ho Ghil(길성호) (검색결과 3 건)
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Goα를 영구적으로 발현하는 세포에서의 신경돌기 성장 억제
길성호(Sung-Ho Ghil),서해영(Haeyoung Suh-Kim) 대한해부학회 2002 Anatomy & Cell Biology Vol.35 No.2
G 단백질은 신경전달물질과 호르몬에 의해서 발생되는 신호를 세포내부로 중개하는 신호전달물질이다. G 단백질 중 Go 는 뇌조직의 막에 상당량 발현되는 단백질이지만, 아직까지 그 기능이 정확히 알려져 있지 않다. 본 연구팀은 선행연구에서 Go의 α 소단위체인 αo를 신경모세포종인 F11세포에 일시적으로 과발현시키면, 신경돌기의 형성에 큰 영향이 있음을 보고한바 있다. 본 연구에서는 이러한 변화가 유전자의 일시적 과발현에 의해서 생기는 비특이 적 현상인지, 아니면 αo에 의해서 일어나는 특이적 현상인지를 규명하기 위하여 F11 세포에 αo를 영구적으로 발현하는 세포를 제작한 후, αo가 지속적으로 발현될 때 신경세포의 분화과정에서의 변화를 조사하였다. αo의 야생형과 돌연변이형을 지속적으로 발현하는 F11 세포를 모두 제작하였다. 각 세포에 dibutyryl cAMP를 처리한 후 신경돌기의 길이를 측정하였다. 정상적인 F11 세포의 경우, 신경돌기의 길이는 57.9±7.0 μm이었으며, 야생형과 돌연변이 αo가 지속적으로 발현된 세포의 경우, 신경돌기의 길이는 각각 34.4±5.1 μm와 30.5±3.6 μm이었다. 이는 정상 F11 세포에 비하여 각각 40.6%, 47.3% 감소한 것으로서, αo가 지속적으로 발현되는 경우에, 일시 과발현되는 경우와 마찬가지로 신경세포의 분화과정 중 특히 신경돌기의 형성과정을 조절한다는 사실을 의미한다. 나아가 두 가지 다른 접근법에 의하여 동일한 결과를 얻었으므로 신경돌기 형성과정의 변화는 αo에 특이적으로 일어남을 알 수 있었다. Heterotrimeric G proteins mediate signals generated by neurotransmitters and hormones. Among G proteins, Go is found in a large quantity in brain and growth cone membranes of neurons. In spite of its abundance in neurons, the role of Go is not fully understood. In the previous study, we showed that transient expression of the α subunit of Go (αo) modulated neurite outgrowth in F11 cells. It is possible that transient transfection may cause transient accumulation of the protein, which itself may alter differentiation process in non-specific manner. In this study, we determined that modulation of neurite outgrowth by αo was specific by evaluating the effect of αo in stably transformed F11 cells. F11 cells stably expressing the wild type αo (αowt) and a constitutively active form of αo (αoQ205L) were established. In normal F11 cells and αo-stable cell lines, the neurite length was measured in the presence of dibutyryl cAMP. In normal F11 cells, the average length of neurites was 57.9±7.0 μm. In αowt- and αoQ205L-expressing cells, the average length were 34.4±5.1 μm 30.5±3.6 μm, respectively. Thus, stable expression of αowt and αoQ205L caused a decrease in neurite outgrowth by 40.6%, 47.3% respectively. This result indicates that modulation of neurite by αo was specific to the function of αo but not due to accumulation of exogenous proteins.
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금강모치 (Rhynchocypris kumgangensis)에서 heat shock protein 70의 클로닝과 수온상승에 의한 발현 변화 분석
임지수 ( Ji Su Im ),길성호 ( Sung Ho Ghil ) 한국물환경학회 2013 한국물환경학회지 Vol.29 No.2
Water temperature is key factor influencing growth and reproduction of fish and its increase give rise to various physiological changes including gene expression. Heat shock protein(Hsp), one of the molecular chaperones, is highly conserved throughout evolution and its expression is induced by various stressors such as temperature, oxidative, physical and chemical stresses. Here, we isolated partial cDNA clones encoding 70 kDa Hsp(Hsp70) and B actin using reverse transcriptase PCR(RT PCR) from gut of Rhynchocypris kumgangensis, a Korean indigenous species and cold water fish, and investigated expression profiles of Hsp70 under an increase of water temperature using B action as an intermal control for RT PCR. Cloned Hsp70 cDNA of R. kumgangensis showed homology to Ctenopharyngodon idella (96%), Hypophthalmichthys molitrix (96%). Danio rerio (93%) and Oncorhynchus mykiss(81%) Hsp70. Cloned B actin cDNA of R. kumgangensis shouwed homology to D. rerio (98%), H, molitrix (97%), C. idella(97%) and O. mykiss(90%) B actin, Both mRNA of Hsp70 and B actin were expressed in gut, brain, and liver in R. kumgangensis. Futhermore, expression of Hsp70, in brain, was highly augmented by an increase of water temperature. These results suggest that Hsp70 mRNA expression level in brain can be used as a biological molecular marker to represent physiological stress against an increase of water temperature.
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Fluorescence-based Assay System for Endocannabinoid Degradation Enzyme, Fatty Acid Amide Hydrolase
Dae-Woong Kim,Gun-Joong Kim,Hae-Jo Kim,Sung-Ho Ghil(길성호) 대한의생명과학회 2010 Biomedical Science Letters Vol.16 No.4
Endogenous cannabinoids (endocannabinoids) display various pharmacological effects including pain control, anti-inflammation, and neuroprotection. The synthesis and release of endocannabinoids are regulated under both physiological and pathological conditions. The main degrading enzyme of endocannabinoid is fatty acid amide hydrolase (FAAH). Therefore we have developed the fluorescence-based assay system for FAAH. We established stable CosM6 cell lines expressing human FAAH. We also synthesized 2-oxo-2H-chromen-7-yl decanoate (DAEC) as a fluorogenic substrate for FAAH. When crude membrane extracts stably expressing FAAH was incubated with DAEC at 25℃, FAAH reacted specifically to DAEC and catalyzes the hydrolysis of DAEC into decanoic acid and highly fluorescent coumarin. Furthermore, the serin hydrolase inhibitor, phenylmethanesulfonylfluoride, inhibited the coumarin release to the reaction buffer in concentration dependent manner. This assay system is suitable for high-throughput screening since this system has simple experimental procedure and measurement method.
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