TRPC6 participates in the regulation of cytosolic basal calcium concentration in murine resting platelets.
Cytosolic-free Ca(2+) plays a crucial role in blood platelet function and is essential for thrombosis and hemostasis. Therefore, cytosolic-free Ca(2+) concentration is tightly regulated in this cell. TRPC6 is expressed in platelets, and an important role for this Ca(2+) channel in Ca(2+) homeostasis has been reported in other cell types. The aim of this work is to study the function of TRPC6 in platelet Ca(2+) homeostasis. The absence of TRPC6 resulted in an 18.73% decreased basal [Ca(2+)]c in resting platelets as compared to control cells. Further analysis confirmed a similar Ca(2+) accumulation in wild-type and TRPC6-deficient mice; however, passive Ca(2+) leak rates from agonist-sensitive intracellular stores were significantly decreased in TRPC6-deficient platelets. Biotinylation studies indicated the presence of an intracellular TRPC6 population, and subcellular fractionation indicated their presence on endoplasmic reticulum membranes. Moreover, the presence of intracellular calcium release in platelets stimulated with 1-oleoyl-2-acetyl-sn-glycerol further suggested a functional TRPC6 population located on the intracellular membranes surrounding calcium stores. However, coimmunoprecipitation assay confirmed the absence of STIM1-TRPC6 interactions in resting conditions. This findings together with the absence of extracellular Mn(2+) entry in resting wild-type platelets indicate that the plasma membrane TRPC6 fraction does not play a significant role in the maintenance of basal [Ca(2+)]c in mouse platelets. Our results suggest an active participation of the intracellular TRPC6 fraction as a regulator of basal [Ca(2+)]c, controlling the passive Ca(2+) leak rate from agonist-sensitive intracellular Ca(2+) stores in resting platelets.
Phycell Group, Department of Physiology, University of Extremadura, Cáceres, 10003, Extremadura, Spain.
Letizia
L
Albarran
Phycell Group, Department of Physiology, University of Extremadura, Cáceres, 10003, Extremadura, Spain. Electronic address: alejandrobe@unex.es.
Alejandro
A
Berna-Erro
Phycell Group, Department of Physiology, University of Extremadura, Cáceres, 10003, Extremadura, Spain.
Natalia
N
Dionisio
Phycell Group, Department of Physiology, University of Extremadura, Cáceres, 10003, Extremadura, Spain.
Pedro C
PC
Redondo
Phycell Group, Department of Physiology, University of Extremadura, Cáceres, 10003, Extremadura, Spain.
Esther
E
Lopez
Phycell Group, Department of Physiology, University of Extremadura, Cáceres, 10003, Extremadura, Spain.
Jose J
JJ
Lopez
Phycell Group, Department of Physiology, University of Extremadura, Cáceres, 10003, Extremadura, Spain.
Gines M
GM
Salido
Blood Bank of Extremadura, Servicio Extremeño de Salud, 06800, Mérida, Extremadura, Spain.
Jose M
JM
Brull Sabate
Phycell Group, Department of Physiology, University of Extremadura, Cáceres, 10003, Extremadura, Spain.
Juan A
JA
Rosado
D000818:Animals; D001792:Blood Platelets / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism*; D003600:Cytosol / Q000378:metabolism; D004075:Diglycerides / Q000494:pharmacology; D006706:Homeostasis; D007425:Intracellular Membranes / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D009363:Neoplasm Proteins / Q000378:metabolism; D000071737:Stromal Interaction Molecule 1; D050052:TRPC Cation Channels / Q000235:genetics / Q000378:metabolism*; D000074287:TRPC6 Cation Channel
D004075:Diglycerides; D008565:Membrane Proteins; D009363:Neoplasm Proteins; C103530:STIM1 protein, human; D000071737:Stromal Interaction Molecule 1; D050052:TRPC Cation Channels; D000074287:TRPC6 Cation Channel; C117032:TRPC6 protein, human; C038237:1-oleoyl-2-acetylglycerol; D002118:Calcium
Calcium; Hemostasis; Platelets; TRPC; Thrombosis