Mitochondrial toxin inhibition of [(3)H]dopamine uptake into rat striatal synaptosomes.
Administration of the mitochondrial inhibitors malonate and 3-nitropropionic acid (3-NP) to rats provides useful models of Huntington's disease. Exposure to these inhibitors has been shown to result in increased extracellular concentrations of striatal dopamine (DA), which is neurotoxic at high concentrations. The cause of this increase is unknown. The purpose of this study was to determine whether mitochondrial inhibition alters dopamine transporter (DAT) function. Striatal synaptosomes were incubated in the presence of several structurally unrelated inhibitors of mitochondrial Complexes I, II, and IV, and [(3)H]DA uptake was measured. Although all of the toxins inhibited [(3)H]DA uptake, there was a large variation in their inhibitory potencies, the rank order being rotenone>>cyanide>azide>3-NP>>malonate. Examination of the kinetic parameters of [(3)H]DA uptake revealed that inhibition was due to a reduction in maximum velocity (V(max)), with no change in affinity (K(m)). The addition of either ATP or of ADP plus P(i) to synaptosomes treated with 3-NP, or of the reactive oxygen species spin trap alpha-phenyl-N-tert-butyl nitrone to synaptosomes exposed to either malonate or cyanide failed to prevent mitochondrial toxin-induced inhibition of DAT function. The lack of effect of high energy substrates or of a free radical scavenger suggests that the mechanism by which extracellular DA is increased by several mitochondrial toxins involves factors other than mitochondrial ATP production or oxidative stress. Taken together, the results suggest that one mechanism whereby mitochondrial toxins increase extracellular concentrations of DA is via interaction with the DAT at a site other than the substrate site, i.e. noncompetitive inhibition of the DAT.
Maragos
William F
WF
Department of Neurology, Kentucky Clinic, Room L-445, University of Kentucky Medical Center, Lexington, KY 40536, USA. maragos@uky.edu
Zhu
Jun
J
Chesnut
M Dathan
MD
Dwoskin
Linda P
LP
D000818:Animals; D001692:Biological Transport; D003342:Corpus Striatum / Q000187:drug effects* / Q000378:metabolism; D004298:Dopamine / Q000378:metabolism*; D008297:Male; D008314:Malonates / Q000494:pharmacology*; D008928:Mitochondria / Q000187:drug effects* / Q000378:metabolism; D009574:Nitro Compounds; D011422:Propionates / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013574:Synaptosomes / Q000187:drug effects* / Q000378:metabolism; D014316:Tritium
D008314:Malonates; D009574:Nitro Compounds; D011422:Propionates; D014316:Tritium; C030290:malonic acid; C015392:3-nitropropionic acid; D004298:Dopamine