Rapid and Sensitive Neurotoxicity Test Based on the Morphological Changes of PC12 Cells with Simple Computer-Assisted Image Analysis

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概要

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• In order to develop a rapid and sensitive bioassay for the screening of chemicals with possible neurotoxicity, a computer-assisted simple image-analysis system was developed to quantify small changes in the specific morphology of the cultured pheochromocytoma cell line, PC12. This cell forms a neuronlike microfibril network(neurites) in response to a nerve growth factor(NGF) stimulation in vitro. Dichrolvos(DDVP) and methylmercury chloride(MMC) were employed as model neurotoxicants. In DDVP treatment, there was no large difference in the ED_<50>s(effective dose that reduces the morphological index by 50%) among the toxicities determined from various morphological indices, but they were significantly lower than those observed by whole-cell-area-based toxicity assay using the hepatoblastoma cell line, Hep G2. In contrast, in MMC treatment, neurite-length-based toxicity was observed as early as 2h, and at 48h this was lower by over three orders of magnitude compared with whole-cell-area-based one (2.06×10^<-7>mM vs. 6.4×10^<-4>mM). These results demonstrate that the developed bioassay using image analysis of nerve-tissue-derived cell morphology allows us to screen possible neurotoxic chemicals very rapidly with highly enhanced sensitivity, particularly for some chemicals that preferentially act on nerve fibers.

• 社団法人日本生物工学会の論文
• 2000-07-25

著者

• Sakai Yasuyuki

  Institute of Industrial Science, University of Tokyo

• SUZUKI MOTOYUKI

  Institute of Industrial Science, University of Tokyo

• Suzuki M

  Institute Of Industrial Science University Of Tokyo

• SAKODA Akiyoshi

  Institute of Industrial Science, University of Tokyo

• SHOJI RYO

  Institute of Industrial Science, University of Tokyo

• MISHIMA YUTAKA

  Institute of Industrial Science, University of Tokyo

• Shoji Ryo

  Institute Of Industrial Science University Of Tokyo

• Sakai Y

  Center For Disease Biology And Integrative Medicine Graduate School Of Medicine University Of Tokyo

• Mishima Y

  Mishima Inst. Dermatological Res. Kobe

• Mishima Yutaka

  Institute Of Industrial Science University Of Tokyo

• Suzuki Motoyuki

  Institute Of Technology And Science The University Of Tokushima

• Sakoda A

  Institute Of Industrial Science University Of Tokyo

• Sakoda Akiyoshi

  Institute Of Industrial Science University Of Tokyo

• Sakai Yasuyuki

  Institute Of Industrial Science (iis) The University Of Tokyo

• Suzuki Motoyuki

  Institute Of Industrial Science University Of Tokyo

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