Effect of t-butyl hydroperoxide on Ca 2+ movement in PC12 pheochromocytoma cells

Cheng Hsien Lu, Warren Su, Yuk Keung Lo, Wei Chung Chen, Wen Neng Chang, Jue Long Wang, Yu Chu Tsai, Ping Yu Lee, Chung Ren Jan*

*Corresponding author for this work

Research output: Contribution to journalJournal Article peer-review

6 Scopus citations


The effect of the oxidant t-butyl hydroperoxide on intracellular free levels of Ca 2+ ([Ca 2+ ]i) in PC12 pheochromocytoma cells was examined by using fura-2 as a fluorescent dye. t-Butyl hydroperoxide induced an increase in [Ca 2+ ] i in a concentration-dependent fashion between 50-250 μM with an EC50 of 100 μM. The [Ca 2+ ] i signal consisted of a slow rise and a sustained phase. The response was decreased by 65% by removal of extracellular Ca 2+ . In Ca 2+ -free medium, pretreatment with 1 μM thapsigargin (an endoplasmic reticulum Ca 2+ pump inhibitor) abolished 150 μM t-butyl hydroperoxide-induced [Ca 2+ ] i increase, and conversely, pretreatment with t-butyl hydroperoxide abrogated thapsigargin-induced [Ca 2+ ] i increase. The 150 μM t-butyl hydroperoxide-induced [Ca 2+ ] i increase in Ca 2+ medium was reduced by 42±5% by pretreatment with 0.1 μM nicardipine but not by 10 μM verapamil, nifedipine, nimodipine or diltiazem, or by 50 μM La 3+ or Ni 2+ . Pretreatment with 10 μM t-butyl hydroperoxide for 40 min did not affect 10 μM ATP-induced [Ca 2+ ] i increase. Together, the results show that t-butyl hydroperoxide induced significant [Ca 2+ ] i increase in PC12 cells by causing store Ca 2+ release from the thapsigargin-sensitive endoplasmic reticulum pool in an inositol 1,4,5-trisphosphate-independent manner and by inducing Ca 2+ influx via a nicardipine-sensitive pathway.

Original languageEnglish
Pages (from-to)51-56
Number of pages6
JournalChinese Journal of Physiology
Issue number2
StatePublished - 2002
Externally publishedYes


  • Fura-2
  • PC12 cells
  • Thapsigargin
  • t-butyl hydroperoxide


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