Calcium dependence of purinergic subtype P2Y₁ receptor modulation of C6 glioma cell migration.

Abstract

We have examined activation of purinergic P2Y₁ receptor-dependent Ca²⁺-signaling pathways in mediating C6 glioma cell migration. The administration of 2-methylthioadenosine 5'-diphosphate (2MeSADP), a selective agonist for P2Y₁R, induced marked increases in patterns of glioma migration in both scratch wound and Boyden chamber assays. Antagonism of P2Y₁R with either the broad spectrum purinergic blocker, pyridoxal-phosphate-6-azophenyl-2',4'-disulfonate (PPADS) or the specific P2Y₁R antagonist, 2'-deoxy-N⁶-methyladenosine-3',5'-bisphosphate (MRS2179), significantly inhibited C6 cell migration. Calcium-sensitive spectrofluorometry showed 2MeSADP stimulation of glioma cells caused a biphasic change in intracellular Ca²⁺ ([Ca²⁺]i). The rapid transient phase was unchanged in Ca²⁺-free solution reflecting a [Ca²⁺]i component due to intracellular stores release subsequent to activation of a metabotropic P2Y subtype receptor. The secondary prolonged phase of [Ca²⁺]i was abolished in Ca²⁺-free solution or in glioma cells treated with the store-operated channel (SOC) blocker, SKF96365. Treatment of glioma with either MRS2179 or PPADS significantly attenuated both the rapid and prolonged phases of [Ca²⁺]i. These results suggest critical roles for activation of P2Y₁R in mediating glioma cell mobility and migration with changes in [Ca²⁺]i contributing as a mechanistic link between activated receptor and functional response. Our findings suggest that pharmacological modulation of metabotropic P2Y₁R-dependent signaling pathways may serve as a novel therapeutic procedure to slow glioma progression.

• Authors: Nattinee Jantaratnotai, James G McLarnon
• Journal: Neuroscience letters (Neurosci Lett) Vol. 497 Issue 2 Pg. 80-4 (Jun 22 2011) ISSN: 1872-7972 [Electronic] Ireland
• PMID: 21540076 (Publication Type: Journal Article)
• Copyright: Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

Chemical References

• Calcium Channel Blockers
• Purinergic P2Y Receptor Agonists
• Purinergic P2Y Receptor Antagonists
• Receptors, Purinergic P2Y1
• Thionucleotides
• methylthio-ADP
• Adenosine Diphosphate
• Calcium

Topics

• Adenosine Diphosphate (analogs & derivatives, pharmacology)
• Animals
• Brain Neoplasms (metabolism, pathology)
• Calcium (metabolism)
• Calcium Channel Blockers (pharmacology)
• Calcium Signaling (drug effects, physiology)
• Cell Line, Tumor
• Cell Movement (drug effects, physiology)
• Glioma (metabolism, pathology)
• Intracellular Fluid (metabolism)
• Ion Channel Gating (drug effects, physiology)
• Neoplasm Invasiveness (pathology)
• Purinergic P2Y Receptor Agonists (pharmacology)
• Purinergic P2Y Receptor Antagonists (pharmacology)
• Rats
• Receptors, Purinergic P2Y1 (drug effects, physiology)
• Thionucleotides (pharmacology)
• Time Factors