title: ADP-evoked phospholipase C stimulation and adenylyl cyclase inhibition in glioma C6 cells occur through two distinct nucleotide receptors, P2Y(1) and P2Y(12).
abstract: In this study we characterized the subtypes of nucleotide P2Y receptors that respond to ADP in glioma C6 cells. Direct visualization of phosphatidylinositol 4,5-bisphosphate at the cell surface revealed that extracellular ADP activates phospholipase C (PLC). Knock-down of P2Y(1) receptor with antisense oligonucleotide, as well as treatment with MRS2179 and pyridoxal-phosphate-6-azophenyl-2',4'-disulfonic acid (P2Y(1) antagonists), attenuates receptor-mediated PLC activity. Adenylyl cyclase inhibition by ADP remains unchanged under these conditions. Reverse transcription-PCR analysis showed that P2Y(12) receptor is expressed in C6 cells. We therefore conclude that, in glioma C6 cells, two P2Y receptor subtypes are present: P2Y(1), coupled to PLC, and P2Y(12), negatively coupled to adenylyl cyclase.
authors.affiliation: Department of Molecular and Cellular Neurobiology, Nencki Institute of Experimental Biology, 3 Pasteur Street, 02-093, Warsaw, Poland.
authors.forename: Rafał
authors.initials: R
authors.lastname: Czajkowski
authors.forename: Lingsheng
authors.initials: L
authors.lastname: Lei
authors.forename: Pawel
authors.initials: P
authors.lastname: Sabała
authors.forename: Jolanta
authors.initials: J
authors.lastname: Barańska
mesh_terms: D000244:Adenosine Diphosphate / Q000378:metabolism; D000067956:Adenylyl Cyclase Inhibitors; D000262:Adenylyl Cyclases / Q000378:metabolism*; D000818:Animals; D004789:Enzyme Activation; D005910:Glioma / Q000201:enzymology*; D008565:Membrane Proteins; D051381:Rats; D018048:Receptors, Purinergic P2 / Q000378:metabolism*; D058472:Receptors, Purinergic P2Y1; D058925:Receptors, Purinergic P2Y12; D014407:Tumor Cells, Cultured; D010738:Type C Phospholipases / Q000378:metabolism*
chemical_list: D000067956:Adenylyl Cyclase Inhibitors; D008565:Membrane Proteins; C546384:P2ry1 protein, rat; C553058:P2ry12 protein, rat; D018048:Receptors, Purinergic P2; D058472:Receptors, Purinergic P2Y1; D058925:Receptors, Purinergic P2Y12; D000244:Adenosine Diphosphate; D010738:Type C Phospholipases; D000262:Adenylyl Cyclases