ATP induces intracellular calcium increases and actin cytoskeleton disaggregation via P2x receptors

Cell Calcium. 2001 May;29(5):299-309. doi: 10.1054/ceca.2000.0194.

Abstract

The consequences of purinoceptor activation on calcium signalling, inositol phosphate metabolism, protein secretion and the actin cytoskeleton were demonstrated in the WRK-1 cell line. Extracellular ATP was used as a secretagogue to induce a rise in intracellular Ca(2+) concentration ([Ca(2+)](i)), acting via P2x purinergic receptors, which causes actin skeleton disaggregation and protein secretion. ATP bound specifically to purinergic receptors, with Ki of 0.8 microM. The magnitude order for binding of different nucleotides was alpha beta-Met-ATP >or= dATPalphaS > ATP >or= ADP > UTP > AMP > suramin. No increase in inositol phosphates (IPs) was observed after ATP application suggesting that the purinergic sites in WRK-1 cells are not of a P2y type. ATP (1-100 microM) caused a concentration-dependent increase in [Ca(2+)](i)(EC(50)= 30 microM). The responses were reproducible without any desensitization over several applications. The response to ATP was abolished when extracellular calcium ([Ca(2+)](e)) was reduced to 100 nM. A non-specific purinergic antagonist, suramin, reversibly inhibited the ATP-response suggesting that ATP is able to bind to P2x purinergic sites to trigger Ca(2+) entry and increase of [Ca(2+)](i). ATP induced a concentration-dependent disaggregation of actin and exocytotic release of proteins both, which were dependent upon [Ca(2+)](e). Similarly, alpha,beta-Met-ATP, a potent P2x agonist also stimulated Ca(2+) mobilization, actin network destructuration, and protein release. In the isolated rat neurohypophysial nerve terminals, ATP was shown to act as a physiological stimulus for vasopressin release via Ca(2+) entry through a P2x receptor [6]. Here, we show that in these nerve terminals, ATP is also able to induce actin disaggregation by a Ca(2+) dependent mechanism. Thus, actin cytoskeleton alterations induced by ATP through activation of P2x receptors could be a prelude to exocytosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / metabolism*
  • Adenosine Triphosphate / metabolism*
  • Animals
  • Binding Sites
  • Calcium / metabolism
  • Calcium Signaling / physiology*
  • Cell Membrane / metabolism
  • Cytoskeleton / metabolism
  • Deoxyadenine Nucleotides / metabolism
  • Inositol Phosphates / metabolism
  • Intracellular Fluid
  • Rats
  • Receptors, Purinergic P2 / metabolism*
  • Thionucleotides / metabolism
  • Tumor Cells, Cultured

Substances

  • Actins
  • Deoxyadenine Nucleotides
  • Inositol Phosphates
  • Receptors, Purinergic P2
  • Thionucleotides
  • 2'-deoxyadenosine 5'-O-(1-thiotriphosphate)
  • Adenosine Triphosphate
  • Calcium