Adenosine A1-receptor stimulation of inositol phospholipid hydrolysis and calcium mobilisation in DDT1 MF-2 cells

Br J Pharmacol. 1992 May;106(1):215-21. doi: 10.1111/j.1476-5381.1992.tb14317.x.


1. The effect of adenosine receptor-stimulation on inositol phospholipid hydrolysis and calcium mobilization has been investigated in the hamster vas deferens smooth muscle cell line DDT1 MF-2. 2. Adenosine receptor stimulation increased the accumulation of total [3H]-inositol phosphates in DDT1 MF-2 cells prelabelled with [3H]-myo-inositol. The rank order of agonist potencies was N6-cyclopentyladenosine greater than 5'-N-ethylcarboxamidoadenosine greater than 2-chloroadenosine greater than adenosine. 3. The response to 2-chloroadenosine was antagonized by the antagonists 8-cyclopentyl-1,3-dipropylxanthine (KD 1.2 nM), PD 115,199 (KD 39 nM) and 8-phenyltheophylline (KD 31 nM). 4. The inositol phosphate response to 2-chloradenosine (10 microM) was not significantly altered when the extracellular Ca2+ ion concentration was reduced from 2.4 mM to 1.2 mM or 0.6 mM. Under calcium-free conditions, however, a reduced but still significant response to 2-chloroadenosine was evident (39 +/- 10% of the response in calcium-containing medium). 5. The 5-lipoxygenase inhibitor AA861 (10 and 100 microM) inhibited the inositol phosphate response to 2-chloroadenosine by 40 +/- 9% and 60 +/- 4% respectively. The cyclo-oxygenase inhibitor, indomethacin, however, was without significant effect at 1 microM. 6. 2-Chloroadenosine stimulated an increase in intracellular free Ca2+ ion concentration in fura-2 loaded DDT1 MF-2 cells in calcium-free medium containing 0.1 mM EGTA, which could be inhibited by the adenosine A1-receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine (0.1 microM). 7. These data suggest that adenosine A1-receptor stimulation results in inositol phospholipid hydrolysis and calcium mobilization from intracellular stores in DDT1 MF-2 cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 2-Chloroadenosine / pharmacology
  • Adenosine / pharmacology
  • Animals
  • Benzoquinones / pharmacology
  • Calcium / metabolism*
  • Cell Line
  • Cricetinae
  • Hydrolysis
  • Indomethacin / pharmacology
  • Lipoxygenase Inhibitors / pharmacology
  • Male
  • Muscle, Smooth / drug effects
  • Muscle, Smooth / metabolism*
  • Phosphatidylinositols / metabolism*
  • Receptors, Purinergic / metabolism*


  • Benzoquinones
  • Lipoxygenase Inhibitors
  • Phosphatidylinositols
  • Receptors, Purinergic
  • 2-Chloroadenosine
  • 2,3,5-trimethyl-6-(12-hydroxy-5,10-dodecadiynyl)-1,4-benzoquinone
  • Adenosine
  • Calcium
  • Indomethacin