Autoregulation in PC12 cells via P2Y receptors: Evidence for non-exocytotic nucleotide release from neuroendocrine cells.

Purinergic Signalling
Simon HusslStefan Boehm

Abstract

Nucleotides are released not only from neurons, but also from various other types of cells including fibroblasts, epithelial, endothelial and glial cells. While ATP release from non-neural cells is frequently Ca(2+) independent and mostly non-vesicular, neuronal ATP release is generally believed to occur via exocytosis. To evaluate whether nucleotide release from neuroendocrine cells might involve a non-vesicular component, the autocrine/paracrine activation of P2Y(12) receptors was used as a biosensor for nucleotide release from PC12 cells. Expression of a plasmid coding for the botulinum toxin C1 light chain led to a decrease in syntaxin 1 detected in immunoblots of PC12 membranes. In parallel, spontaneous as well as depolarization-evoked release of previously incorporated [(3)H]noradrenaline from transfected cells was significantly reduced in comparison with the release from untransfected cells, thus indicating that exocytosis was impaired. In PC12 cells expressing the botulinum toxin C1 light chain, ADP reduced cyclic AMP synthesis to the same extent as in non-transfected cells. Likewise, the enhancement of cyclic AMP synthesis either due to the blockade of P2Y(12) receptors or due to the degradation of extracellular neucle...Continue Reading

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Citations

Jun 26, 2013·Purinergic Signalling·Geoffrey Burnstock, Francesco Di Virgilio
Aug 29, 2009·The European Journal of Neuroscience·Ilia A YambolievVioleta N Mutafova-Yambolieva
Nov 20, 2008·Clinical and Experimental Allergy : Journal of the British Society for Allergy and Clinical Immunology·M A M Willart, B N Lambrecht
Jun 3, 2014·Pharmacology & Therapeutics·Violeta N Mutafova-Yambolieva, Leonie Durnin
Jan 14, 2010·Science Signaling·Ross Corriden, Paul A Insel

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Methods Mentioned

BETA
biosensor
transfection
electrophoresis

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