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TheScientificWorldJOURNAL (ISSN 1537-744X)

Article Details

Title:

Organotypic Cultures as a Model of Parkinson´s Disease. A Twist to an Old Model

Authors:

Stahl, Katja ; Skare, Oivind ; Torp, Reidun

Journal:

TheScientificWorldJOURNAL

Year:

2009

Volume:

Page Range:

811-821

Article Type:

Methods Paper

Handling Editor:

Rudy Tanzi

Domains:

Neuroscience , Aging , Neurology , Methods & Protocols

DOI:

10.1100/tsw.2009.68

cell death - compounds - cultures - induction - internal control - mesencephalon - microelectrode - neurons - new application - ohda - pd - precise quantification - refinement - representativeness - tissue slices - tissue surface - toxicity - toxin - vm - western blots

Synopsis:

Modeling Parkinson´s disease in organotypic cultures has traditionally been done by adding dopamine-selective toxic compounds in the culture media. This method induces dopaminergic degeneration but does not account for natural variation among slices. We present a new unilateral application technique that accounts for the variation and enables precise quantification of the cell loss due to the toxin.

Keywords:

Parkinson’s disease, organotypic cultures, 6-hydroxydopamine (6-OHDA), confocal microscopy

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Abstract

Organotypic cultures from the ventral mesencephalon (VM) are widely used to model Parkinson’s disease (PD). In this method, neurotoxic compounds have traditionally been applied to the media to induce a uniform dopaminergic (DAergic) cell death in the tissue slices, regardless of the variation existing among slices. This study demonstrates a refinement of the toxic induction technique. We show that unilateral application of 6-hydroxydopamine (6-OHDA) at the tissue surface by means of a microelectrode causes a precisely localized cell death that closely resembles an in vivo stereotactic model. This technique introduces an internal control that accounts for variation between slices and enables a precise quantification of the cell loss due to the toxin in use. We characterized organotypic VM cultures in terms of effects of 6-OHDA toxicity and number of DAergic neurons as judged by immunofluorescence and Western blots. Our findings illustrate that this new application technique greatly improves the representativeness of organotypic cultures as a model for PD.We characterized organotypic VM cultures in terms of effects of 6-OHDA toxicity and number of DAergic neurons as judged by immunofluorescence and Western blots. Our findings illustrate that this new application technique greatly improves the representativeness of organotypic cultures as a model for PD.

TheScientificWorldJOURNAL (ISSN 1537-744X)

Article Details