Acute versus long-term effects of 6-hydroxydopamine on oxidative stress and dopamine depletion in the striatum of mice

Varcin, Mustafa; Bentea, Eduard; Mertens, Birgit; Van den Haute, Chris; Baekelandt, Veerle; Michotte, Yvette; Sarre, Sophie

doi:10.1016/J.JNEUMETH.2011.07.004

Title

Acute versus long-term effects of 6-hydroxydopamine on oxidative stress and dopamine depletion in the striatum of mice

Author

Varcin, Mustafa

Bentea, Eduard

Mertens, Birgit

Van den Haute, Chris

Baekelandt, Veerle

Michotte, Yvette

Sarre, Sophie

Abstract

Oxidative stress is one of the mechanisms which may be important in the pathogenesis of Parkinson's disease. In the current study, the effects of 6-hydroxydopamine (6-OHDA) perfusion on hydroxyl radical formation in the mouse striatum were investigated using the in vivo salicylate trapping microdialysis technique. The latter uses salicylate as a trapping agent for hydroxyl radicals with formation of 2,3-dihydroxybenzoic acid (2,3-DHBA), which is measured by HPLC. Two different approaches of the technique were validated in mice. First, perfusion of the trapping agent salicylate (1 mM) via the probe in combination with 6-OHDA (5 mu M) was used to screen for radical scavenging properties of compounds in mice. Alternatively, striatal administration of 6-OHDA in a concentration known to induce nigrostriatal denervation (1 mM), without the trapping agent, allowed to maximally challenge the neuronal microenvironment and as such to investigate both its acute and long-term effects. In the first method, as expected, glutathione (GSH) (1.5 mM) prevented the 6-OHDA-induced increase in 2,3-DHBA levels. In the second method, GSH prevented the hydroxyl radical formation, while depletion of GSH with 2-cyclohexen-1-one (CHO) resulted in significantly higher 2,3-DHBA levels than when 6-OHDA was perfused alone. Three weeks after the local 6-OHDA perfusion, the total striatal dopamine (DA) and dihydroxyphenylacetic acid (DOPAC) content were reduced by 30%, compared to the intact striatum, accompanied by a reduction in striatal tyrosine hydroxylase (TH) immunoreactive (ir) nerve terminals. This suggests that the second method can be used to determine the acute as well as the long-term effects of 6-OHDA in the mouse striatum. (C) 2011 Elsevier B.V. All rights reserved.

Language

English

Source (journal)

Journal of neuroscience methods. - Amsterdam

Source (book)

13th International Conference on In Vivo Methods, SEP 12-16, 2010, Brussels, BELGIUM

Publication

Amsterdam : Elsevier science bv , 2011

ISSN

0165-0270

DOI

10.1016/J.JNEUMETH.2011.07.004