Pyridostigmine bromide, chlorpyrifos, and DEET combined Gulf War exposure insult depresses mitochondrial function in neuroblastoma cells

J Biochem Mol Toxicol. 2021 Dec;35(12):e22913. doi: 10.1002/jbt.22913. Epub 2021 Sep 15.


Gulf War Illness (GWI) is defined by the Centers for Disease Control and Prevention (CDC) as a multi-symptom illness having at least one symptom from two of three factors, which include: fatigue, mood-cognition problems, and musculoskeletal disorders. The cluster of long-term symptoms is unique to military personnel from coalition countries including United States, Australia, and the United Kingdom that served in Operation Desert Storm from 1990 to 1991. Reporting of these symptoms is much lower among soldiers deployed in other parts of the world like Bosnia during the same time period. The exact cause of GWI is unknown, but combined exposure to N,N-diethyl-m-toluamide (DEET), organophosphates like chlorpyrifos (CPF), and pyridostigmine bromide (PB), has been hypothesized as a potential mechanism. Mitochondrial dysfunction is known to occur in most neurodegenerative diseases that share symptoms with GWI and has therefore been implicated in GWI. Although exposure to these and other toxicants continues to be investigated as potential causes of GWI, their combined impact on mitochondrial physiology remains unknown. In this study, the effects of combined GWI toxicant exposure on mitochondrial function were determined in a commonly used and readily available immortalized cell line (N2a), whose higher rate of oxygen consumption resembles that of highly metabolic neurons in vivo. We report that combined exposure containing pesticide CPF 71 μM, insect repellants DEET 78 μM, and antitoxins PB 19 μM, causes profound mitochondrial dysfunction after a 4-h incubation resulting in decreased mitochondrial respiratory states in the absence of proapoptotic signaling, proton leak, or significant increase in reactive oxygen species production.

Keywords: ATP; Gulf War Illness; mitochondria; oxygen consumption; reactive oxygen species.

MeSH terms

  • Adenosine Triphosphate / biosynthesis
  • Animals
  • Apoptosis / drug effects
  • Cell Line, Tumor
  • Chlorpyrifos / toxicity*
  • DEET / toxicity*
  • Humans
  • Mice
  • Mitochondria / drug effects*
  • Mitochondria / metabolism
  • Neuroblastoma / pathology*
  • Oxygen Consumption / drug effects
  • Persian Gulf Syndrome*
  • Protein Kinases / metabolism
  • Pyridostigmine Bromide / toxicity*
  • Signal Transduction / drug effects
  • War Exposure*


  • DEET
  • Adenosine Triphosphate
  • Protein Kinases
  • Chlorpyrifos
  • Pyridostigmine Bromide