Free radical determinants of endogenous and amphetamine-enhanced neurodegenerative disease
Subtitle: prostaglandin H synthase-catalyzed free radical formation, reactive oxygen species-mediated oxidative DNA damage and glucose-6-phosphate dehydrogenase-catalyzed neuroprotection.
Published: 2004
Language: eng
Number of pages: 359
Description: The amphetamine analogs 3,4-methylenedioxymethamphetamine (MDMA, "Ecstasy"), 3,4-methylenedioxyamphetamine (MDA) and methamphetamine (METH) are bioactivated by prostaglandin H synthase (PHS) to free radical intermediates that initiate the formation of reactive oxygen species (ROS), which oxidatively damage cellular macromolecules. Using electron paramagnetic resonance spectrometry, the mechanism of free radical formation for MDMA, MDA and METH were evaluated in an  in vitro system with purified PHS-1, and the free radical spin trapping agent, alpha-phenyl-N-t-butylnitrone. MDMA, MDA and METH all demonstrated PHS-catalyzed, time- and concentration-dependent formation of a carbon-centered- and/or nitrogen-centered free radical intermediate. Moreover, these amphetamines were stereoselective in causing oxidative DNA damage, which was blocked by the PHS inhibitor, eicosatetraynoic acid. Similarly in vivo, these amphetamines were stereoselective in mediating oxidative DNA damage, dopaminergic neurodegeneration, and permanent motor coordination deficits. These in vivo effects were inhibited by a single pretreatment with the PHS inhibitor aspirin (with MDA), or reduced in a gene dose-dependent manner in PHS-1 knockout mice (with MDMA). In addition to the neurodegenerative effects in adult brain, administration of a single dose of METH to pregnant mice in mid- (organogenesis) or late gestation caused increased DNA damage in fetal brain and permanent neurodevelopmental deficits, evidenced by postnatal impairment of motor coordination.These results implicate PHS-catalyzed free radical formation and ROS-mediated oxidative damage in xenobiotic- and endobiotic-initiated neurodegeneration, and indicate a neuroprotective role for GOD, thus providing a novel hypothesis potentially relevant to the mechanisms and risk factors for neurodegeneration associated with amphetamines, as well as with aging.PHS also bioactivated various endogenous neurotransmitters and precursors in vitro leading to DNA oxidation. Moreover, aged -/- PHS-1-deficient knockout mice had significantly decreased oxidative DNA damage in various brain regions compared to wild-type PHS-1 normal controls, with PHS-1 heterozygotes exhibiting intermediary risk. Thus, oxidative DNA damage initiated in a PHS-dependent fashion by endogenous substrates may contribute to the neurodegeneration associated with aging. The neurodegenerative potential of endogenous ROS was confirmed in mutant mice with a deficiency in the antioxidative enzyme glucose-6-phosphate dehydrogenase (G6PD), as evidenced by increased oxidative DNA damage and neurodegenerative changes in -/- G6PD-deficient mice.
Notes: Adviser:  Peter George Wells.

Thesis (Ph.D.)--University of Toronto, 2004.

Electronic version licensed for access by U. of T. users.

Source: Dissertation Abstracts International, Volume: 65-10, Section: B, page: 5117.