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Effect of 6-hydroxydopamine (6-OHDA) on proliferation of glial cells in the rat cortex and striatum: evidence for de-differentiation of resident astrocytes

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Abstract

Reactive astrogliosis is the universal response to any brain insult. It is characterized by cellular hypertrophy, up-regulation of the astrocyte marker glial fibrillary acidic protein (GFAP), and proliferation. The source of these proliferating cells is under intense debate. Progenitor cells derived from the subventricular zone (SVZ), cells positive for chondroitin sulfate proteoglycan (NG2+), and de-differentiated astrocytes have been proposed as the origin of proliferating cells following injury. We have analyzed the effect of intraventricular-applied 6-hydroxydopamine (6-OHDA) on the proliferation and morphology of astrocytes in rat cortex and striatum by means of immunohistochemistry and confocal laser microscopy. At 4 days post-lesion, GFAP expression increased markedly. A subpopulation of the GFAP+ cells co-expressed Ki-67, indicating that these cells were proliferating. To investigate whether these cells (1) arose from migrating SVZ progenitor cells, (2) derived from NG2+ progenitor cells, or (3) de-differentiated from resident astrocytes, we studied the expression of the migration marker doublecortin (Dcx), the oligodendrocyte progenitor marker NG2, and the progenitor markers Nestin and Pax6. The proliferating Ki-67+ cells co-expressed Nestin and Pax6, whereas no co-expression of Ki-67 with NG2 or the migration marker Dcx was observed. Thus, resident astrocytes de-differentiate, in response to the intraventricular application of 6-OHDA, to a phenotype resembling radial glia cells, which represent transient astrocyte precursors during development. An understanding of the mechanisms of the de-differentiation of mature astrocytes might be useful for designing new approaches to cell therapy in neurodegenerative diseases such as Parkinson’s disease.

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Abbreviations

6-OHDA:

6-hydroxydopamine

ABC:

avitin-biotin peroxidase complex

DAB:

3,3’-diaminobenzidine

Dcx:

doublecortin

GFAP:

glial fibrillary acidic protein

MPTP:

1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine

NG2:

chondroitin sulfate proteoglycan

Pax6:

paired box gene 6

PBS:

phosphate-buffered saline

SVZ:

subventricular zone

TH:

tyrosine hydroxylase

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Acknowledgements

The authors thank Ulrich Mattheus, Elke Maier, and Mihnea Nicolescu for excellent technical assistance and Stephan Grissmer for critical reading of the manuscript.

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Authors and Affiliations

  1. Institute of Anatomy, Department of Cellular Neurobiology, University of Tübingen, Oesterbergstrasse 3, 72074, Tübingen, Germany

    Britta Wachter, Hans-Jochen Wagner & Eva Kueppers

  2. Institute of Neurobiology, Department of Neuropharmacology, Tübingen, Germany

    Sonja Schürger, Jens Rolinger & Andreas von Ameln-Mayerhofer

  3. Center of Neurology, Hertie Institute for Clinical Brain Research, Tübingen, Germany

    Sonja Schürger, Jens Rolinger & Daniela Berg

  4. Graduate School for Cellular and Molecular Neuroscience, Eberhard Karls University, Tübingen, Germany

    Britta Wachter

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  1. Britta Wachter
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  2. Sonja Schürger
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Correspondence to Eva Kueppers.

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Wachter, B., Schürger, S., Rolinger, J. et al. Effect of 6-hydroxydopamine (6-OHDA) on proliferation of glial cells in the rat cortex and striatum: evidence for de-differentiation of resident astrocytes. Cell Tissue Res 342, 147–160 (2010). https://doi.org/10.1007/s00441-010-1061-x

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