title: Adenylate cyclase 5 coordinates the action of ADP, P2Y1, P2Y13 and ATP-gated P2X7 receptors on axonal elongation.
abstract: In adult brains, ionotropic or metabotropic purinergic receptors are widely expressed in neurons and glial cells. They play an essential role in inflammation and neurotransmission in response to purines secreted to the extracellular medium. Recent studies have demonstrated a role for purinergic receptors in proliferation and differentiation of neural stem cells although little is known about their role in regulating the initial neuronal development and axon elongation. The objective of our study was to investigate the role of some different types of purinergic receptors, P2Y1, P2Y13 and P2X7, which are activated by ADP or ATP. To study the role and crosstalk of P2Y1, P2Y13 and P2X7 purinergic receptors in axonal elongation, we treated neurons with specific agonists and antagonists, and we nucleofected neurons with expression or shRNA plasmids. ADP and P2Y1-GFP expression improved axonal elongation; conversely, P2Y13 and ATP-gated P2X7 receptors halted axonal elongation. Signaling through each of these receptor types was coordinated by adenylate cyclase 5. In neurons nucleofected with a cAMP FRET biosensor (ICUE3), addition of ADP or Blue Brilliant G, a P2X7 antagonist, increased cAMP levels in the distal region of the axon. Adenylate cyclase 5 inhibition or suppression impaired these cAMP increments. In conclusion, our results demonstrate a crosstalk between two metabotropic and one ionotropic purinergic receptor that regulates cAMP levels through adenylate cyclase 5 and modulates axonal elongation triggered by neurotropic factors and the PI3K-Akt-GSK3 pathway.
authors.affiliation: Department of Molecular, Cellular and Developmental Neurobiology, Instituto Cajal, CSIC, Madrid,  Spain.
authors.forename: Ana
authors.initials: A
authors.lastname: del Puerto
authors.forename: Juan-Ignacio
authors.initials: JI
authors.lastname: Díaz-Hernández
authors.forename: Mónica
authors.initials: M
authors.lastname: Tapia
authors.forename: Rosa
authors.initials: R
authors.lastname: Gomez-Villafuertes
authors.forename: María José
authors.initials: MJ
authors.lastname: Benitez
authors.forename: Jin
authors.initials: J
authors.lastname: Zhang
authors.forename: María Teresa
authors.initials: MT
authors.lastname: Miras-Portugal
authors.forename: Francisco
authors.initials: F
authors.lastname: Wandosell
authors.forename: Miguel
authors.initials: M
authors.lastname: Díaz-Hernández
authors.forename: Juan José
authors.initials: JJ
authors.lastname: Garrido
mesh_terms: D000244:Adenosine Diphosphate / Q000378:metabolism* / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000378:metabolism*; D000262:Adenylyl Cyclases / Q000378:metabolism*; D000818:Animals; D001369:Axons / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D048430:Cell Shape / Q000187:drug effects; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000378:metabolism; D020868:Gene Silencing; D038362:Glycogen Synthase Kinase 3 / Q000378:metabolism; D000071679:Glycogen Synthase Kinase 3 beta; D057809:HEK293 Cells; D006624:Hippocampus / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D051379:Mice; D051058:Oncogene Protein v-akt / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D018048:Receptors, Purinergic P2 / Q000378:metabolism; D058486:Receptors, Purinergic P2X7 / Q000378:metabolism*; D058472:Receptors, Purinergic P2Y1 / Q000378:metabolism*; D012394:Rosaniline Dyes / Q000494:pharmacology
chemical_list: C440036:P2RY13 protein, human; D018048:Receptors, Purinergic P2; D058486:Receptors, Purinergic P2X7; D058472:Receptors, Purinergic P2Y1; D012394:Rosaniline Dyes; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D000242:Cyclic AMP; D019869:Phosphatidylinositol 3-Kinases; D000071679:Glycogen Synthase Kinase 3 beta; D051058:Oncogene Protein v-akt; D038362:Glycogen Synthase Kinase 3; D000262:Adenylyl Cyclases; C419510:adenylyl cyclase type V; C004692:coomassie Brilliant Blue