Summary
We have examined the activation of a phospholipase C signal transduction pathway by a B2-bradykinin receptor in the human astrocytoma cell line D384 and how this influences D1-dopamine receptor stimulated cyclic AMP accumulation. Addition of bradykinin to D384 cells resulted in a concentration-dependent (10−11 − 10−6 M) increase in the accumulation of [3H]inositol phosphates and a similar concentration-dependent transient increase in specific [3H]β-phorbol-12,13-dibutyrate binding which is indicative of translocation of protein kinase C from the cytosol to the membrane. Changes in intracellular Ca2+ of single cells, measured using the fluorescent indicator dye fura-2, indicated that bradykinin produced a rapid, but transient, increase in intracellular calcium. The Ca2+ response was largely independent of extracellular Ca2+ supporting the idea that receptor activation leads to mobilization of Ca2+ from intracellular stores. However, extracellular Ca2+ was required for a response to a rechallenge with bradykinin. The bradykinin B2-receptor agonist kallidin increased cytosolic Ca 2+ in a similar manner to bradykinin. The Ca2+ response to bradykinin could be partially reduced in the presence of the B2-receptor antagonist [d-Arg10-Hyp,d-Phe7,β-(2-Thienyl)-Ala5,8]-bradykinin, whereas the B1-receptor agonists (Des-Arg9]-bradykinin and [Des-Arg10]-kallidin were ineffective. Bradykinin was also found to attenuate dopamine stimulated cyclic AMP accumulation in D384 cells, at similar concentrations previously observed to stimulate the phospholipase C signal transduction pathway, in the presence of the phosphodiesterase inhibitor, rolipram. In contrast, no attenuation was observed in the presence of the phosphodiesterase inhibitor 1-isobutyl 3-methylxanthine, although the level of dopamine stimulated cyclic AMP observed was lower than in the presence of rolipram. Furthermore, the effect of bradykinin could be mimicked by a calcium ionophore, but not a phorbol ester. These data suggest that D384 cells express a B2 bradykinin receptor coupled to polyphosphoinositide-specific phospholipase C. Activation of this receptor results in elevated levels of cytosolic Ca2+ and to a reduction of D1-dopamine receptor stimulated cyclic AMP accumulation.
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Correspondence to A. J. Balmforth at the above address
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Balmforth, A.J., Parkinson, F.E., Altiok, N. et al. Identification of a B2-bradykinin receptor linked to phospholipase C and inhibition of dopamine stimulated cyclic AMP accumulation in the human astrocytoma cell line D384. Naunyn-Schmiedeberg's Arch Pharmacol 346, 303–310 (1992). https://doi.org/10.1007/BF00173543
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DOI: https://doi.org/10.1007/BF00173543