A novel method for analyzing mitochondrial movement: inhibition by paclitaxel in a pheochromocytoma cell model

J Mol Neurosci. 2009 Mar;37(3):254-62. doi: 10.1007/s12031-008-9129-8. Epub 2008 Jul 18.

Abstract

A method was developed to assess mitochondrial movement in the living cell that is dependent, in part, on microtubule and/or associating protein interactions. The leader sequence from cytochrome-c was used to drive DsRed2 fluorescent proteins to accumulate in the mitochondria, thus enabling to follow mitochondrial (cytochrome-c's) movement. For calculating the percentage of mitochondrial movement, an image-processing program was used (ImageJ). Paclitaxel, an antitumor agent, is a potent microtubule-stabilizing agent that increases the stability of tubulin polymers, inhibiting mitosis and mitochondrial activity in dividing cells. Here, we tested whether paclitaxel inhibits mitochondrial movement in pheochromocytoma cells (a neuronal model, when tested in a differentiated state). While a 2-day exposure to paclitaxel resulted in cellular toxicity (measured as inhibition of mitochondrial activity), 2-3 h exposure to paclitaxel were sufficient to inhibit mitochondrial movement as assessed in 10-20-s imaging sessions in living cells. Mitotracker deep-red staining validated the staining obtained with DsRed2-cytochrome-c and identified intact mitochondria. Results showed a significant paclitaxel dose-dependent inhibition of mitochondrial movement. This new method should enable further assessment of microtubule-interacting drugs and other cytoskeletal components for their potential influence of mitochondrial movement as a test for activity and side effects.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adrenal Gland Neoplasms / metabolism*
  • Animals
  • Antineoplastic Agents, Phytogenic / pharmacology*
  • Cell Line, Tumor
  • Fluorescent Dyes / metabolism
  • Mitochondria* / drug effects
  • Mitochondria* / metabolism
  • PC12 Cells / cytology
  • PC12 Cells / drug effects
  • PC12 Cells / physiology
  • Paclitaxel / pharmacology*
  • Pheochromocytoma / metabolism*
  • Rats

Substances

  • Antineoplastic Agents, Phytogenic
  • Fluorescent Dyes
  • Paclitaxel