Selection of Molecular Biomarkers Relevant to Abnormal Behaviors of Medaka Fish (Oryzias latipes) Caused by Diazinon

다이아지논에 의해 야기된 송사리의 이상행동 연관 분자생물지표의 선발

  • Koh, Sung-Cheol (Division of Civil and Environmental Engineering, Korea Maritime University) ;
  • Shin, Sung-Woo (Division of Civil and Environmental Engineering, Korea Maritime University) ;
  • Cho, Hyun-Duk (Department of Biological Sciences, Pusan Naitonal University) ;
  • Chon, Tae-Soo (Department of Biological Sciences, Pusan Naitonal University) ;
  • Kim, Jong-Sang (Department of Animal Biotechnology, Kyungpook Naitonal University) ;
  • Lee, Sung-Kyu (Korea Institute of Toxicology)
  • 고성철 (한국해양대학교 건설환경공학부) ;
  • 신성우 (한국해양대학교 건설환경공학부) ;
  • 조현덕 (부산대학교 생명과학부) ;
  • 전태수 (부산대학교 생명과학부) ;
  • 김정상 (경북대학교 동물공학과) ;
  • 이성규 (안전성평가연구소)
  • Published : 2009.12.31

Abstract

본 연구의 목적은 다이아지논(Diazinon; O, O-diethyl O-[6-methyl-2 (1-methylethyl)-4-pyrimidinyl] phosphorothioate)에 노출된 모델 생물체(송사리)의 행동변화와 관련된 분자생물학적 기전 규명을 통하여 비정상적 행동의 모니터링을 위한 생물지표(biomarker)를 개발하는데 있다. 이를 위해 우선 suppression subtractive hybridization (SSH) 및 DNA microarray 기법을 활용하여 다양한 유전자를 스크리닝하였다. 다이아지논에 노출시킨 송사리에서 발현의 차이가 나는 상향 조절된 유전자 97개 (알려지지 않은 유전자 27개 포함)와 하향 조절된 유전자 99개 (알려지지 않은 유전자 60개 포함)를 동정 하였고 이들 중 이상행동과 관련되는 것으로 보이는 유전자 10개 (상향조절 5개, 하향조절 5개)를 선발하였다. 이들 중에서 primer 제작이 잘된 beta-1, Orla C3-1, parvalbumin 및 apolipoprotein E을 선발하여 그 유전자 발현을 real-time PCR 기법을 사용하여 정량적으로 모니터링 하였다. Orla C3-1, parvalbumin 및 apolipoprotein E는 고농도의 다이아지논 처리(1000 ppb; 24 h)에서 그 발현이 억제됨이 관찰되었다. 다이아지논 처리 시 신경질환 (알츠하이머 병 및 다운신드롬)에 관련된 apolipoprotein E와 근육세포의 유연화에 작용하는 parvalbumin 등의 발현억제는 송사리의 인지능력 교란 및 근육세포의 경직 등을 각각 유도하여 송사리의 비정상적 행동을 야기하는 것으로 판단되었다. 따라서 이들 생물지표는 신경독성물질에 의한 송사리 및 기타 어류의 이상행동의 변화의 감지에 활용될 수 있을 것으로 사료된다.

Keywords

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