Methoxychlor inhibits brain mitochondrial respiration and increases hydrogen peroxide production and CREB phosphorylation.
The organochlorine insecticide methoxychlor (mxc) is an established reproductive toxicant that affects other systems including the central nervous system (CNS), possibly by mechanisms involving oxidative stress. This study tested the hypothesis that mxc inhibits brain mitochondrial respiration, resulting in increased production of reactive oxygen species (ROS). Oxygen electrode measurements of mitochondrial respiration and Amplex Red measurements of H(2)O(2) production were performed with rat brain mitochondria exposed in vitro to mxc (0-10 microg/ml) and with brain mitochondria from mice chronically exposed in vivo to mxc (0-64 mg/kg/day) for 20 days by intraperitoneal injection. In vitro mxc exposure inhibited ADP-dependent respiration (state 3) using both complex I- and II-supported substrates. Similarly, state 3 respiration was inhibited following in vivo mxc exposure using complex I substrates. H(2)O(2) production was stimulated after in vitro mxc treatment in the presence of complex I substrates, but not in mitochondria isolated from in vivo mxc-treated mice. Because previous studies demonstrated a relationship between oxidative stress and CREB phosphorylation, we also tested the hypothesis that mxc elevates phosphorylated CREB (pCREB) in mitochondria. Enzyme-linked immunosorbent assay (ELISA) measurements demonstrated that pCREB immunoreactivity was elevated by in vitro mxc exposure in the presence or absence of respiratory substrates, indicating that stimulation of H(2)O(2) production is not necessary for this effect. These multiple effects of mxc on mitochondria may play an important role in its toxicity, particularly in the CNS.
Schuh
Rosemary A
RA
Department of Anesthesiology, University of Maryland School of Medicine, Baltimore, 21201, USA.
Kristián
Tibor
T
Gupta
Rupesh K
RK
Flaws
Jodi A
JA
Fiskum
Gary
G
D000818:Animals; D001921:Brain / Q000187:drug effects* / Q000378:metabolism; D019069:Cell Respiration / Q000187:drug effects; D017362:Cyclic AMP Response Element-Binding Protein / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D005260:Female; D006861:Hydrogen Peroxide / Q000378:metabolism*; D007274:Injections, Intraperitoneal; D007306:Insecticides / Q000633:toxicity*; D008297:Male; D008731:Methoxychlor / Q000633:toxicity*; D051379:Mice; D008815:Mice, Inbred Strains; D008928:Mitochondria / Q000187:drug effects* / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D018384:Oxidative Stress / Q000187:drug effects*; D010101:Oxygen Consumption / Q000187:drug effects; D010766:Phosphorylation; D051381:Rats; D017207:Rats, Sprague-Dawley
C492004:Creb1 protein, mouse; D017362:Cyclic AMP Response Element-Binding Protein; D007306:Insecticides; D009419:Nerve Tissue Proteins; D006861:Hydrogen Peroxide; D008731:Methoxychlor