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First published online January 25, 2006
doi: 10.1242/10.1242/dev.02233

Development 133, 675-684 (2006)
Published by The Company of Biologists 2006
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Extracellular nucleotide signaling in adult neural stem cells: synergism with growth factor-mediated cellular proliferation

Santosh K. Mishra1, Norbert Braun1, Varsha Shukla1, Marc Füllgrabe1, Christof Schomerus2, Horst-Werner Korf2, Christian Gachet3, Yukio Ikehara4, Jean Sévigny5, Simon C. Robson6 and Herbert Zimmermann1,*

1 Biocenter, J.W. Goethe-University, AK Neurochemistry, Frankfurt am Main, Germany.
2 Institute of Anatomy II, Medical School, J.W. Goethe-University, Frankfurt am Main, Germany.
3 INSERM U.311, EFS-Alsace, Strasbourg, France.
4 Department of Cell Biology, Fukuoka University School of Medicine, Japan.
5 Centre de Recherche en Rhumatologie et Immunologie, Sainte-Foy, (Québec), Canada.
6 Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA 02215, USA.

* Author for correspondence (e-mail: h.zimmermann{at}cns.uni-frankurt.de)

Accepted 1 December 2005

We have previously shown that the extracellular nucleoside triphosphate-hydrolyzing enzyme NTPDase2 is highly expressed in situ by stem/progenitor cells of the two neurogenic regions of the adult murine brain: the subventricular zone (type B cells) and the dentate gyrus of the hippocampus (residual radial glia). We explored the possibility that adult multipotent neural stem cells express nucleotide receptors and investigated their functional properties in vitro. Neurospheres cultured from the adult mouse SVZ in the presence of epidermal growth factor and fibroblast growth factor 2 expressed the ecto-nucleotidases NTPDase2 and the tissue non-specific isoform of alkaline phosphatase, hydrolyzing extracellular ATP to adenosine. ATP, ADP and, to a lesser extent, UTP evoked rapid Ca2+ transients in neurospheres that were exclusively mediated by the metabotropic P2Y1 and P2Y2 nucleotide receptors. In addition, agonists of these receptors and low concentrations of adenosine augmented cell proliferation in the presence of growth factors. Neurosphere cell proliferation was attenuated after application of the P2Y1-receptor antagonist MRS2179 and in neurospheres from P2Y1-receptor knockout mice. In situ hybridization identified P2Y1-receptor mRNA in clusters of SVZ cells. Our results infer nucleotide receptor-mediated synergism that augments growth factor-mediated cell proliferation. Together with the in situ data, this supports the notion that extracellular nucleotides contribute to the control of adult neurogenesis.

Key words: Adult neurogenesis, ATP, Ecto-Nucleotidase, Neurosphere, P2 receptor, Subventricular zone


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