Stimulation of phospholipase C activity by norepinephrine, t-ACPD and bombesin in LA-N-2 cells

The release of [3H]inositol phosphates from myo-[3H]inositol-prelabeled LA-N-2 cells was measured in the presence of beta-adrenoceptor, metabotropic glutamate and bombesin agonists. Norepinephrine and isoproterenol increased the formation of [3H]inositol phosphates in a dose-dependent manner, with an EC50 of 100 microM for norepinephrine and an EC50 of 5 microM for isoproterenol. These stimulations were abolished by propranolol, a beta-adrenoceptor antagonist, with an IC50 in the range of 50-55 microM for both norepinephrine and isoproterenol. The stimulation of [3H]inositol phosphate appearance occurred with varying concentrations of trans-1-amino-1,3-cyclopentanedicarboxylic acid (t-ACPD), a metabotropic glutamate receptor agonist. This release of [3H] inositol phosphates was blunted by its antagonist, 2-amino-3-phosphonopropionic acid (AP-3). Bombesin and neuromedin-B, a bombesin-like peptide, also increased the appearance of [3H]inositol phosphates. This was blunted by the antagonist [Tyr4, D-Phe12] bombesin. The appearance of [3H]inositol phosphates stimulated by t-ACPD was coupled through a cholera toxin-sensitive G-protein and the bombesin-stimulated appearance of [3H]inositol phosphates was coupled through a pertussis toxin-sensitive G-protein. The norepinephrine-stimulated appearance of [3H]inositol phosphates was toxin insensitive. The stimulation of the [3H]inositol phosphate appearance by these three agonists was protein kinase and Ca2+ independent.