Rotenone-induced apoptosis and role of calcium: A study on Neuro-2a cells

Supriya Swarnkar, Poonam Goswami, Pradeep Kumar Kamat, Sonam Gupta, Ishan K. Patro, Sarika Singh, Chandishwar Nath

Research output: Contribution to journalArticlepeer-review

42 Scopus citations


Rotenone causes cytotoxicity in astrocytic cell culture by glial activation, which is linked to free radical generation. The present study is an investigation to explore whether rotenone could also cause cellular toxicity in mouse neuroblastoma cells (Neuro-2a) under treatment similar to astroglial cells. The effect of rotenone (0.1, 1, and 10 μM) onmitochondrial dehydrogenase enzyme activity by MTT reduction assay, PI uptake, total reactive oxygen species (ROS)/superoxide levels, nitrite levels, extent of DNA damage (by comet assay), and nuclear morphological alteration by Hoechst staining was studied. Caspase-3 and Ca2+/calmodulin-dependent protein kinase II (CaMKIIα) gene expression was determined to evaluate the apoptotic cell death and calcium kinase, respectively. Calcium level was estimated fluorometrically using fura-2A stain. Rotenone decreased mitochondrial dehydrogenase enzyme activity and generated ROS, superoxide, and nitrite. Rotenone treatment impaired cell intactness and nuclear morphology as depicted by PI uptake and chromosomal condensation of Neuro-2a cells, respectively. In addition, rotenone resulted in increased intracellular Ca+2 level, caspase-3, and CaMKIIα expression. Furthermore, co-exposure of melatonin (300 μM), an antioxidantto cell culture, significantly suppressed the rotenone-induced decreased mitochondrial dehydrogenase enzyme activity, elevated ROS and RNS. However, melatonin was found ineffective to counteract rotenone-induced increased PI uptake, alteredmorphological changes, DNA damage, elevated Ca+2, and increased expression of caspase-3 and CaMKIIα. The study indicates that intracellular calcium rather than oxidative stress is a major factor for rotenoneinduced apoptosis in neuronal cells.

Original languageEnglish (US)
Pages (from-to)1387-1397
Number of pages11
JournalArchives of Toxicology
Issue number9
StatePublished - Sep 2012
Externally publishedYes


  • Apoptosis
  • DNA damage
  • Melatonin
  • Neuronal cells
  • Reactive oxygen species
  • Rotenone

ASJC Scopus subject areas

  • Toxicology
  • Health, Toxicology and Mutagenesis


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