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The Journal of Neuroscience, December 10, 2003, 23(36):11444-11452

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Cellular/Molecular
Reduced Expression of P2Y₁ Receptors in Connexin43-Null Mice Alters Calcium Signaling and Migration of Neural Progenitor Cells

Eliana Scemes,¹ Nathalie Duval,² and Paolo Meda³

¹Department of Neuroscience, Albert Einstein College of Medicine, Bronx, New York 10461, ²Rétrovirus et Transfert Génétique, Institute Pasteur, 75015 Paris, France, and ³Department de Morphologie, Centre Medical Universitaire, CH-1211 Geneva 4, Switzerland

Glial calcium signals play important roles during CNS development. Calcium transients induced by ATP, acting on purinergic receptors, stimulate DNA synthesis, increase astrocytic and neural stem cell proliferation, and are prominent during the differentiation of radial glia. We have shown previously that expression of P2Y receptors in astrocytes is altered when connexin43 (Cx43) is downregulated. To evaluate the consequences of Cx43 deletion on calcium signaling during neural progenitor development, studies were performed on neurospheres derived from embryonic striatum. After adhesion, cells migrating from wild-type (WT) and Cx43-null neurospheres displayed spontaneous calcium oscillations. Such activity was blunted by apyrase, 2'-deoxy-N⁶-methyladenosine 3',5'-bisphosphate (MRS-2179), and suramin, suggesting that ATP released by neural cells acts on purinergic receptors to induce calcium oscillations. The amplitudes of Ca²⁺ transients induced by P2Y but not P2X receptor agonists were larger in WT than in Cx43-null progenitors, suggesting that these two cell populations express different P2 receptors. Suramin, a nonselective P2 receptor antagonist, and MRS-2179, a P2Y₁ receptor-selective antagonist, reduced the proliferation rate and the migration of WT progenitor cells to levels similar to those of Cx43-null cells. Conversely, exogenous expression of P2Y₁ receptors in Cx43-null cells restored their migration pattern to levels seen in WT progenitors. However, treatment with P2 receptor antagonists did not alter the ratio of nestin to GFAP expression in WT neural progenitors. These data show that altered autocrine-paracrine communication attributable to reduced levels of P2Y₁ receptors in neural progenitor cells lacking Cx43 affects proliferation and migration but not cell differentiation during early CNS development.

Key words: P2 receptors; calcium oscillations; neurospheres; progenitors; gap junction; connexin

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Received Aug 8, 2003; revised October 28, 2003; accepted October 28, 2003.

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