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$Na^+/H^+$ exchanger와 $HCO_3^-$ transporter에 의한 흰쥐 타액선 선세포내 pH 조절
박동범,서정택,손흥규,이종갑,Park, Dong-Bum,Seo, Jeong-Taeg,Sohn, Heung-Kyu,Lee, Jong-Gap 대한소아치과학회 1998 大韓小兒齒科學會誌 Vol.25 No.2
Intracellular pH (pHi) plays an important role in the regulation of cellular processes by influencing the acitivity of various enzymes in cells. Therefore, almost every type of mammalian cell possesses an ability to regulate its pHi. One of the most prominent mechanisms in the regulation of pHi is $Na^+/H^+$ exchanger. This exchanger has been known to be activated when cells are stimulated by the binding of agonist to the muscarinic receptors. Therefore, the aims of this study were to compare the rates of $H^+$ extrusion through $Na^+/H^+$ exchanger before and during muscarinic stimulation and to investigate the possible existence of $HCO_3^-$ transporter which is responsible for the continuous supply of $HCO_3^-$ ion to saliva. Acinar cells were isolated from the rat mandibular salivary glands and loaded with pH-sensitive fluoroprobe, 2', 7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein(BCECF), for 30min at room temperature. Cells were attached onto the coverglass in the perfusion chamber and the changes in pHi were measured on the iverted microscope using spectrofluorometer. 1. By switching the perfusate from $HCO_3^-$-free to $HCO_3^-$-buffered solution, pHi decreased by $0.39{\pm}0.02$ pH units followed by a slow increase at an initial rate of $0.04{\pm}0.007$ pH units/min. The rate of pHi increase was reduced to $0.01{\pm}0.002$ pH units/min by the simultaneous addition of 1 mM amiloride and $100{\mu}M$ DIDS. 2. An addition and removal of $NH_4^+$ caused a decrease in pHi which was followed by an increase in pHi. The increase of pHi was almost completely blocked by 1mM amiloride in $HCO_3^-$-free perfusate which implied that the pHi increase was entired dependent on the activation of $Na^+/H^+$ exchanger in $HCO_3^-$-free condition. 3. An addition of $10{\mu}M$ carbachol increased the initial rate of pHi recovery from $0.16{\pm}0.01$ pH units/min to $0.28{\pm}0.03pH$ units/min. 4. The initial rate of pHi decrease induced by 1mM amiloride was also increased by the exposure of the acinar cells to $10{\mu}M$ carbachol ($0.06{\pm}0.008pH$ unit/min) compared with that obtained before carbachol stimulation ($0.03{\pm}0.004pH$ unit/min). 5. The intracellular buffering capacity ${\beta}1$ was $14.31{\pm}1.82$ at pHi 7.2-7.4 and ${\beta}1$ increased as pHi decreased. 6. The rate of $H^+$ extrusion through $Na^+/H^+$ exchanger was greatly enhanced by the stimulation of the cells with $10{\mu}M$ carbachol and there was an alkaline shift in the activity of the exchanger. 7. An intrusion mechanism of $HCO_3^-$ was identified in rat mandibular salivary acinar cells. Taken all together, I observed 3-fold increase in $Na^+/H^+$ exchanger by the stimulation of the acinar cells with $10{\mu}M$ carbachol at pH 7.25. In addition, I have found an additional mechanism for the regulation of pHi which transported $HCO_3^-$ into the cells.
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고성덕(S.D. Ko),천명철(M.C. Chon),김만중(M.J. Kim),윤주용(J.Y. Yoon) 한국해양환경·에너지학회 2011 한국해양환경·에너지학회 학술대회논문집 Vol.2011 No.11
해양오염사고에 의한 기름 유출은 대부분 선박에서 발생하고 있으며 오염 행위자를 밝히지 못하면 피해당사자의 보상문제 뿐만아니라 사고초기에 방제전략 수립 등이 어렵다. 더욱이 선박은 이동성 및 국제항행 특수성이 있어 오염사고 주변해역을 지나간 수백척의 선박을 짧은 시간안에 조사하는 것은 매우 어렵다. 따라서 오염행위자를 적발하기 위해서는 정확한 유출유의 유종 파악과 유출시간 추정을 통해 조사대상을 압축하여 정밀조사하는 것이 효과적이다. 우리나라에서는 이러한 문제를 해결하기 위해 국내에 도입되는 모든 원유, 제품유에 대한 유지문D/B를 구축 하여 활용하고 있으나 해상에 유출된 기름이 주변해역의 기상, 수온 및 경과시간 등 자연조건변화에 따라 물리화학적 특성이 변화한 경우에는 정확한 유종파악에 많은 시간이 소요되며 유출시간을 추정하는 것이 매우 제한적이다. 본 연구에서는 경시변화된 기름의 형광분광광도계를 이용한 감식ㆍ분석방법과 다양한 해역조건에 따라 미국 해양대기청(NOAA)의 ADIOS 2를 활용한 유출시간 추정 방법 등 효율적인 기름 감식ㆍ분석시스템에 대해 연구하였다. Generally oil spill incidents come from vessel. If it happened, especially in case of unknown oil spill incidents, it is difficult to ferret out criminals and compensation for victim grows all the more serious. Besides, It is much difficult to find criminal’s vessel among many vessels. Therefore, it is important to analyse the kind of spilled oil and spilled time of oil because that is one of useful method how to be able to summarize research subjects. KCG(Korea Coast Guard) makes full use of oil fingerprint D/B about crude oil, petroleum oil imported from overseas. But, because the characteristics of spilled oil is changed by weather, temperature and weathering time etc, it is difficult to analyse the oil sample and estimate to spilled time, too. In this study, we introduced how to forensic identify weathering oil by using spectrofluorometer and we studied about oil analysing system by using ADIOS 2 program to be made by NOAA(National Oceanic and Atmospheric Administration).
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Na+/H+ exchanger와 HCO-₃Transporter에 의한 흰쥐 타액선 선세포내 pH 조절
서정택,박동범,손흥규,이종갑 大韓小兒齒科學會 1998 大韓小兒齒科學會誌 Vol.25 No.2
Intracellular pH (pHi) plays an important role in the regulation of cellular processes by influencing the activity of various enzymes in cells. Therefore, almost every type of mammalian cell possesses an ability to regulate its pHi. One of the most prominent mechanisms in the regulation of pHi is Na+/H+ exchanger. This exchanger has been known to be activated when cells are stimulated by the binding of agonist to the muscarinic receptors. Therefore, the aims of this study were to compare the rates of H+ extrusion through Na+/H+ exchanger before and during muscarinic stimulation and to investigate the possible existence of HCO-₃ transporter which is responsible for the continuous supply of HCO-₃ ion to saliva. Acinar cells were isolated from the rat mandibular salivary glands and loaded with pH-sensitive fluoroprobe, 2', 7' -bis(2-carboxyethyl)-5(6)-carboxyfluorescein(BCECF), for 30min at room temperature. Cells were attached onto the coverglass in the perfusion chamber and the changes in pHi were measured on the iverted microscope using spectrofluormeter. 1. By switching the perfusate from HCO-₃-free to HCO-₃-buffered solution, pHi decreased by 0.39±0.02 pH units followed by a slow increase at an initial rate of 0.04±0.007 pH units/min. the rate of pHi increase was reduced to 0.01±0.002 pH units/min by the simultaneous addition of 1 mM amilorede and 100μM DIDS. 2. An addition and removal of NH+₄ caused a decrease in pHi which was followed by an increase in pHi. The increase of pHi was almost completely blocked by 1mM amiloride in HCO-₃-free perfusate which implied that the pHi increase was entired dependent on the activation of Na+/H+ exchanger in HCO-₃=free condition. 3. An addition of 10μM carbachol increased the initial rate of pHi recovery from 0.16±0.01 pH units/min to 0.28±0.03pH units/min. 4. The initial rate of pHi decrease induced by 1mM amilorede was also increased by the exposure of the acinar cells 10μM carbachol(0.06±0.008pH unit/min) compared with that obtained before carbachol sitmulation (0.03±0.004pH unit/min). 5. The intracellular buffering capacity β1 was 14.31±1.82 at pHi 7.2-7.4 and β1 increased as pHi decreased. 6. The rate of H+ extrusion through Na+/H+ exchanger was greatly enhanced by the stimulation of the cells with 10μM carbachol and there was an alkaline shift in the activity of the exchanger. 7. An intrusion mechanism of HCO-₃ was identified in rat mandibular salivary acinar cells. Taken all together, I observed 3-ford increased in Na+/H+ exchanger by the stimulation of the acinar cells with 10μM carbachol at pH 7.25. In addition, I have found an additional mechanism for the regulation of pHi which transported HCO-₃ into the cells.
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Lysosome based toxic detection in Saccharomyces cerevisiae using novel portable fluorometer
Ngoc-Tu Nguyen,안지영,방승혁,김동환,김덕민,김양훈,민지호 대한독성 유전단백체 학회 2016 Molecular & cellular toxicology Vol.12 No.2
Lysosome is an organelle in the cell, commonly used as biomonitoring tool in environmental pollution. In previous studies, the lysosomal proteome profiling in yeast was analyzed in response to different toxic chemicals, such as sodium meta-arsenite and tetracycline. A recombinant yeast contained the specific biomarker found in the previous studies was constructed for toxic detection. We evaluated green fluorescent intensity of the recombinant yeast exposed to toxic chemicals with dose dependent using by a portable kit designed by our laboratory. The results confirmed the potential of both the yeast and the portable kit to detect each of toxic chemical such as heavy metals, pesticides, or pharmaceuticals at an optimal dose that the intensity of fluorescent protein reached peak at. Therefore, the recombinant yeast and portable kit play an important role to detect various toxic chemicals.
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