Activation of P2Y1 receptor triggers two calcium signaling pathways in bone marrow erythroblasts

Eur J Pharmacol. 2006 Mar 18;534(1-3):30-8. doi: 10.1016/j.ejphar.2006.01.010. Epub 2006 Feb 20.

Abstract

In this study, we describe the presence of P2 receptor subtypes and Ca2+ signaling in erythroblasts. ATP and ADP produced a biphasic increase of intracellular Ca2+ concentration ([Ca2+]i), with an initial transient phase followed by a sustained phase. Reverse transcription polymerase chain reaction (RT-PCR) showed the expression of P2Y1, P2Y2 and P2Y12. The selective P2Y1 receptor antagonist 2'-deoxy-N6-methyl-adenosine-3',5'-diphosphate (MRS2179) and the G(i) protein inhibitor pertussis toxin blocked Ca2+ increase. The initial transient [Ca2+]i increase phase was sensitive to the 1,4,5-inositol trisphosphate (IP3) receptor blocker 2-aminoethoxy-diphenylborate (2-APB), while the sustained phase was sensitive to the protein kinase C (PKC) inhibitor 2-[1-(3-dimethylaminopropyl)-1H-indol-3-yl]-3-(1H-indol-3-yl)-maleimide (GF109203X) and calcium calmodulin kinase II (CaMKII) inhibitor 1-[N,O-bis(5-isoquinolinesulfonyl)-N-methyl-L-tyrosyl]-4-phenylpiperazine (KN-62). In addition, the PKC activator phorbol-12,13-dibutyrate (PDBu) produced increase of [Ca2+]i. Flow cytometry analysis showed the expression of Ca2+-dependent PKC alpha, betaI, gamma and phospho-CaMKII. These results suggest that the activation of the P2Y1 receptor triggers two different [Ca2+]i increase pathways, one IP3-dependent and the other kinase-dependent.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / analogs & derivatives
  • 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / pharmacology
  • Adenosine Diphosphate / analogs & derivatives
  • Adenosine Diphosphate / pharmacology
  • Adenosine Triphosphate / pharmacology
  • Animals
  • Bone Marrow Cells / drug effects
  • Bone Marrow Cells / metabolism
  • Boron Compounds / pharmacology
  • Calcium Channels / drug effects
  • Calcium Channels / metabolism
  • Calcium Signaling*
  • Calcium-Calmodulin-Dependent Protein Kinase Type 2
  • Calcium-Calmodulin-Dependent Protein Kinases / antagonists & inhibitors
  • Calcium-Calmodulin-Dependent Protein Kinases / metabolism
  • Dose-Response Relationship, Drug
  • Erythroblasts / drug effects
  • Erythroblasts / metabolism*
  • Female
  • Indoles / pharmacology
  • Inositol 1,4,5-Trisphosphate Receptors
  • Maleimides / pharmacology
  • Mice
  • Mice, Inbred C57BL
  • Protein Kinase C / antagonists & inhibitors
  • Protein Kinase C / metabolism
  • Protein Kinase Inhibitors / pharmacology
  • RNA, Messenger / metabolism
  • Receptors, Cytoplasmic and Nuclear / drug effects
  • Receptors, Cytoplasmic and Nuclear / metabolism
  • Receptors, Purinergic P2 / drug effects
  • Receptors, Purinergic P2 / genetics
  • Receptors, Purinergic P2 / metabolism*
  • Receptors, Purinergic P2Y1

Substances

  • Boron Compounds
  • Calcium Channels
  • Indoles
  • Inositol 1,4,5-Trisphosphate Receptors
  • Maleimides
  • N(6)-methyl-2'-deoxyadenosine 3',5'-diphosphate
  • P2ry1 protein, mouse
  • Protein Kinase Inhibitors
  • RNA, Messenger
  • Receptors, Cytoplasmic and Nuclear
  • Receptors, Purinergic P2
  • Receptors, Purinergic P2Y1
  • Adenosine Diphosphate
  • KN 62
  • 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine
  • Adenosine Triphosphate
  • 2-aminoethoxydiphenyl borate
  • Protein Kinase C
  • Calcium-Calmodulin-Dependent Protein Kinase Type 2
  • Calcium-Calmodulin-Dependent Protein Kinases
  • bisindolylmaleimide I