title: Alpha-synuclein overexpression increases cytosolic catecholamine concentration.
abstract: Dysregulation of dopamine homeostasis and elevation of the cytosolic level of the transmitter have been suggested to underlie the vulnerability of catecholaminergic neurons in Parkinson's disease. Because several known mutations in alpha-synuclein or overexpression of the wild-type (WT) protein causes familial forms of Parkinson's disease, we investigated possible links between alpha-synuclein pathogenesis and dopamine homeostasis. Chromaffin cells isolated from transgenic mice that overexpress A30P alpha-synuclein displayed significantly increased cytosolic catecholamine levels as measured by intracellular patch electrochemistry, whereas cells overexpressing the WT protein and those from knock-out animals were not different from controls. Likewise, catechol concentrations were higher in L-DOPA-treated PC12 cells overexpressing A30P or A53T compared with those expressing WT alpha-synuclein, although the ability of cells to maintain a low cytosolic dopamine level after L-DOPA challenge was markedly inhibited by either protein. We also found that incubation with low-micromolar concentrations of WT, A30P, or A53T alpha-synuclein inhibited ATP-dependent maintenance of pH gradients in isolated chromaffin vesicles and that the WT protein was significantly less potent in inducing the proton leakage. In summary, we demonstrate that overexpression of different types of alpha-synuclein disrupts vesicular pH and leads to a marked increase in the levels of cytosolic catechol species, an effect that may in turn trigger cellular oxyradical damage. Although multiple molecular mechanisms may be responsible for the perturbation of cytosolic catecholamine homeostasis, this study provides critical evidence about how alpha-synuclein might exert its cytotoxicity and selectively damage catecholaminergic cells.
authors.affiliation: Department of Neurology, Columbia University Medical Center, New York, New York 10032, USA.
authors.forename: Eugene V
authors.initials: EV
authors.lastname: Mosharov
authors.forename: Roland G W
authors.initials: RG
authors.lastname: Staal
authors.forename: Jordi
authors.initials: J
authors.lastname: Bové
authors.forename: Delphine
authors.initials: D
authors.lastname: Prou
authors.forename: Anthonia
authors.initials: A
authors.lastname: Hananiya
authors.forename: Dmitriy
authors.initials: D
authors.lastname: Markov
authors.forename: Nathan
authors.initials: N
authors.lastname: Poulsen
authors.forename: Kristin E
authors.initials: KE
authors.lastname: Larsen
authors.forename: Candace M H
authors.initials: CM
authors.lastname: Moore
authors.forename: Matthew D
authors.initials: MD
authors.lastname: Troyer
authors.forename: Robert H
authors.initials: RH
authors.lastname: Edwards
authors.forename: Serge
authors.initials: S
authors.lastname: Przedborski
authors.forename: David
authors.initials: D
authors.lastname: Sulzer
mesh_terms: D000818:Animals; D002395:Catecholamines / Q000378:metabolism*; D002478:Cells, Cultured; D019439:Chromaffin Cells / Q000378:metabolism*; D003600:Cytosol / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000502:physiology; D051379:Mice; D008822:Mice, Transgenic; D016716:PC12 Cells; D051381:Rats; D051844:alpha-Synuclein / Q000378:metabolism*
chemical_list: D002395:Catecholamines; D051844:alpha-Synuclein