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J. Biol. Chem., Vol. 265, Issue 18, 10320-10326, 06, 1990
L Vallar, C Muca, M Magni, P Albert, J Bunzow, J Meldolesi and O Civelli
Dopaminergic D2 receptors are widely regarded as typical inhibitory
receptors, as they both inhibit adenylyl cyclase and decrease the cytosolic
free Ca2+ concentration ([Ca2+]i) by activating K+ channels. A D2 receptor
has recently been cloned (Bunzow, J. R., Van Tol, H. H. M., Grandy, D. K.,
Albert, P., Salon, J., Christie, M. D., Machida, C. A., Neve, K. A., and
Civelli, O. (1988) Nature 336, 783-787) and expressed in two different cell
lines, pituitary GH4C1 cells and Ltk- fibroblasts, where it has been shown
to induce inhibition of adenylyl cyclase. We have investigated the
additional effector systems coupled to this receptor. The responses
observed in the two cells lines, which express similar levels of receptors
(0.5-1 x 10(5)/cell), were surprisingly different. In GH4C1 cells D2
receptors failed to affect phosphoinositide hydrolysis and induced a
decrease of [Ca2+]i. This latter effect appears to be mediated by
hyperpolarization, most likely due to the activation of K+ channels. In
striking contrast, in Ltk- fibroblasts the D2 receptor induced a rapid
stimulation of inositol(1,4,5)-trisphosphate (+73% at 15 s) followed by the
other inositol phosphates, and an immediate increase of [Ca2+]i due to both
Ca2+ mobilization from internal stores and influx from the extracellular
medium. In both GH4C1 and Ltk- cells, the D2 receptor response was mediated
by G protein(s) sensitive to pertussis toxin. The increases of inositol
trisphosphate and [Ca2+]i observed in Ltk- cells required dopamine
concentrations only slightly higher than those inhibiting adenylyl cyclase
(EG50 = 25, 29, and 11 nM, respectively) and were comparable in magnitude
to the responses induced by the endogenous stimulatory receptor agonists,
thrombin and ATP. The results demonstrate that in certain cells D2
receptors are efficiently coupled to the stimulation of phosphoinositide
hydrolysis. The nature of receptor responses appears therefore to depend on
the specific properties not only of the receptor molecule but also of the
cell type in which it is expressed.
Differential coupling of dopaminergic D2 receptors expressed in different cell types. Stimulation of phosphatidylinositol 4,5- bisphosphate hydrolysis in LtK- fibroblasts, hyperpolarization, and cytosolic-free Ca2+ concentration decrease in GH4C1 cells
Department of Pharmacology, University of Milano, Italy.
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