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A Galvanotaxis Assay for Analysis of Neural Precursor Cell Migration Kinetics in an Externally Applied Direct Current Electric Field
Published on: October 13, 2012
The morphological and molecular changes of brain cells exposed to direct current electric field stimulation
Simon J Pelletier1, Marie Lagacé1, Isabelle St-Amour1
1Centre de Recherche du CHU de Québec, Axe Neuroscience, Québec, QC, Canada (Mr Pelletier, Ms Lagacé, Drs St-Amour, Arsenault, Cisbani, and Cicchetti); Département de Psychiatrie et Neurosciences, Université Laval, Québec, QC, Canada (Drs Lévesque and Cicchetti); Centre de recherche de l'Institut Universitaire en Santé Mentale de Québec, Québec, QC, Canada (Ms Chabrat and Dr Lévesque); Laboratory of Canada Research Chair in Cognitive Neuroscience, Centre Interdisciplinaire de Recherche en Réadaptation et Intégration Sociale, Centre de Recherche de l'Institut Universitaire en Santé Mentale de Québec, Université Laval, Canada (Dr Fecteau); Berenson-Allen Center for Noninvasive Brain Stimulation, Beth Israel Deaconess Medical Center, Harvard Medical School, Cambridge, MA (Dr Fecteau).
Background:
The application of low-intensity direct current electric fields has been experimentally used in the clinic to treat a number of brain disorders, predominantly using transcranial direct current stimulation approaches. However, the cellular and molecular changes induced by such treatment remain largely unknown.
Methods:
Here, we tested various intensities of direct current electric fields (0, 25, 50, and 100V/m) in a well-controlled in vitro environment in order to investigate the responses of neurons, microglia, and astrocytes to this type of stimulation. This included morphological assessments of the cells, viability, as well as shape and fiber outgrowth relative to the orientation of the direct current electric field. We also undertook enzyme-linked immunosorbent assays and western immunoblotting to identify which molecular pathways were affected by direct current electric fields.
Results:
In response to direct current electric field, neurons developed an elongated cell body shape with neurite outgrowth that was associated with a significant increase in growth associated protein-43. Fetal midbrain dopaminergic explants grown in a collagen gel matrix also showed a reorientation of their neurites towards the cathode. BV2 microglial cells adopted distinct morphological changes with an increase in cyclooxygenase-2 expression, but these were dependent on whether they had already been activated with lipopolysaccharide. Finally, astrocytes displayed elongated cell bodies with cellular filopodia that were oriented perpendicularly to the direct current electric field.
Conclusion:
We show that cells of the central nervous system can respond to direct current electric fields both in terms of their morphological shape and molecular expression of certain proteins, and this in turn can help us to begin understand the mechanisms underlying the clinical benefits of direct current electric field.
