title: ADP-Induced Ca2+Signaling and Proliferation of Rat Ventricular Myofibroblasts Depend on Phospholipase C-Linked TRP Channels Activation Within Lipid Rafts.
abstract: Nucleotides released during heart injury affect myocardium electrophysiology and remodeling through P2 purinoceptors activation in cardiac myofibroblasts. ATP and UTP endorse [Ca2+]iaccumulation and growth of DDR-2/α-SMA-expressing myofibroblasts from adult rat ventricles via P2Y4and P2Y2receptors activation, respectively. Ventricular myofibroblasts also express ADP-sensitive P2Y1, P2Y12, and P2Y13receptors as demonstrated by immunofluorescence confocal microscopy and western blot analysis, but little information exists on ADP effects in these cells. ADP (0.003-3 mM) and its stable analogue, ADPßS (100 μM), caused fast [Ca2+]itransients originated from thapsigargin-sensitive internal stores, which partially declined to a plateau sustained by capacitative Ca2+entry through transient receptor potential (TRP) channels inhibited by 2-APB (50 μM) and flufenamic acid (100 μM). Hydrophobic interactions between Gq/11-coupled P2Y purinoceptors and TRP channels were suggested by prevention of the ADP-induced [Ca2+]iplateau following PIP2depletion with LiCl (10 mM) and cholesterol removal from lipid rafts with methyl-ß-cyclodextrin (2 mM). ADP [Ca2+]itransients were insensitive to P2Y1, P2Y12, and P2Y13receptor antagonists, MRS2179 (10μM), AR-C66096 (0.1 μM), and MRS2211 (10μM), respectively, but were attenuated by suramin and reactive blue-2 (100 μM) which also blocked P2Y4receptors activation by UTP. Cardiac myofibroblasts growth and type I collagen production were favored upon activation of MRS2179-sensitive P2Y1receptors with ADP or ADPßS (30 μM). In conclusion, ADP exerts a dual role on ventricular myofibroblasts: [Ca2+]itransients are mediated by fast-desensitizing P2Y4receptors, whereas the pro-fibrotic effect of ADP involves the P2Y1receptor activation. Data also show that ADP-induced capacitative Ca2+influx depends on phospholipase C-linked TRP channels opening in lipid raft microdomains. J. Cell. Physiol. 232: 1511-1526, 2017. © 2016 Wiley Periodicals, Inc.
authors.affiliation: Laboratório de Farmacologia e Neurobiologia, Centro de Investigação Farmacológica e Inovação Medicamentosa (MedInUP), Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto (ICBAS-UP), Porto, Portugal.
authors.forename: Mariana
authors.initials: M
authors.lastname: Certal
authors.affiliation: Laboratório de Farmacologia e Neurobiologia, Centro de Investigação Farmacológica e Inovação Medicamentosa (MedInUP), Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto (ICBAS-UP), Porto, Portugal.
authors.forename: Adriana
authors.initials: A
authors.lastname: Vinhas
authors.affiliation: Laboratório de Farmacologia e Neurobiologia, Centro de Investigação Farmacológica e Inovação Medicamentosa (MedInUP), Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto (ICBAS-UP), Porto, Portugal.
authors.forename: Aurora
authors.initials: A
authors.lastname: Barros-Barbosa
authors.affiliation: Laboratório de Farmacologia e Neurobiologia, Centro de Investigação Farmacológica e Inovação Medicamentosa (MedInUP), Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto (ICBAS-UP), Porto, Portugal.
authors.forename: Fátima
authors.initials: F
authors.lastname: Ferreirinha
authors.affiliation: Laboratório de Farmacologia e Neurobiologia, Centro de Investigação Farmacológica e Inovação Medicamentosa (MedInUP), Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto (ICBAS-UP), Porto, Portugal.|Departamento de Química, Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto (ICBAS-UP), Porto, Portugal.
authors.forename: Maria Adelina
authors.initials: MA
authors.lastname: Costa
authors.affiliation: Laboratório de Farmacologia e Neurobiologia, Centro de Investigação Farmacológica e Inovação Medicamentosa (MedInUP), Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto (ICBAS-UP), Porto, Portugal.
authors.forename: Paulo
authors.initials: P
authors.lastname: Correia-de-Sá
mesh_terms: D000244:Adenosine Diphosphate / Q000031:analogs & derivatives / Q000494:pharmacology*; D000375:Aging; D000818:Animals; D002118:Calcium / Q000378:metabolism; D020013:Calcium Signaling / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D024042:Collagen Type I / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects; D005260:Female; D006352:Heart Ventricles / Q000166:cytology*; D057927:Hydrophobic and Hydrophilic Interactions; D015640:Ion Channel Gating / Q000187:drug effects*; D008297:Male; D021962:Membrane Microdomains / Q000378:metabolism*; D058628:Myofibroblasts / Q000187:drug effects / Q000378:metabolism*; D011493:Protein Kinase C / Q000378:metabolism; D017208:Rats, Wistar; D058470:Receptors, Purinergic P2Y / Q000378:metabolism; D019831:Sodium-Calcium Exchanger / Q000378:metabolism; D050051:Transient Receptor Potential Channels / Q000378:metabolism*; D010738:Type C Phospholipases / Q000378:metabolism*
chemical_list: D024042:Collagen Type I; D058470:Receptors, Purinergic P2Y; D019831:Sodium-Calcium Exchanger; D050051:Transient Receptor Potential Channels; D000244:Adenosine Diphosphate; D011493:Protein Kinase C; D010738:Type C Phospholipases; D002118:Calcium