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Inhibition of the T-type Ca2+ current by the dopamine D1 receptor in rat adrenal glomerulosa cells: requirement of the combined action of the G betagamma protein subunit and cyclic adenosine 3',5'-monophosphate

Mol Endocrinol. 1997 Apr;11(4):503-14. doi: 10.1210/mend.11.4.9910.

Abstract

Modulation of ionic Ca2+ currents by dopamine (DA) could play a pivotal role in the control of steroid secretion by the rat adrenal glomerulosa cells. In the present study, we report that DA decreases the T-type Ca2+ current amplitude in these cells. The use of pharmacological agonists and antagonists reveals that this effect is mediated by activation of the D1-like receptors. Modulation by cAMP is complex inasmuch as preincubation of the cells with 8-Br-cAMP or the specific adenylyl cyclase inhibitor, 2',3'-dideoxyadenosine, have no effect per se, but prevent the DA-induced inhibition. The inhibitory effect of DA was abolished by addition of GDPbetaS to the pipette medium but not by pertussis toxin. If a cell is dialyzed with medium containing G alpha(s)-GDP, the inhibitory effect is reduced and cannot be recovered by the addition of GTPgammaS, indicating that the alpha(s) is not involved, but rather the betagamma-subunit. Indeed, DA-induced inhibition was mimicked by G betagamma in the pipette and 8-Br-cAMP in the bath. Similarly, G betagamma release from the activation of the AT1 receptor of angiotensin II did affect the current amplitude only in the presence of 8-Br-cAMP in the bath. The mitogen-activated protein kinase cascade, which can be activated by receptors coupled to Gs, was not involved as shown by the lack of activation of p42mapk by DA and the absence of effect of the mitogen-activated protein kinase inhibitor, PD 098059, on the DA-induced inhibition. Because the binding of G betagamma-subunits to various effectors involves the motif QXXER, we therefore tested the effect of the QEHA peptide on the inhibition of the T-type Ca2+ current induced by DA. The peptide, added to the medium pipette (200 microM), abolished the effect of DA. We conclude that the presence of the G betagamma and an increase in cAMP concentration are both required to inhibit the T-type Ca2+ current in rat adrenal glomerulosa cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenylyl Cyclases / chemistry
  • Adenylyl Cyclases / pharmacology
  • Animals
  • Calcium Channel Blockers / metabolism*
  • Calcium Channels / metabolism*
  • Calcium-Calmodulin-Dependent Protein Kinases / metabolism
  • Cyclic AMP / metabolism*
  • Dopamine / pharmacology
  • Dopamine Agonists / pharmacology
  • Dopamine Antagonists / pharmacology
  • Enzyme Inhibitors / pharmacology
  • Female
  • Flavonoids / pharmacology
  • GTP-Binding Protein alpha Subunits, Gs / metabolism*
  • Guanosine 5'-O-(3-Thiotriphosphate) / pharmacology
  • Guanosine Diphosphate / analogs & derivatives
  • Guanosine Diphosphate / pharmacology
  • Phosphorylation
  • Rats
  • Receptors, Dopamine D1 / metabolism*
  • Second Messenger Systems
  • Thionucleotides / pharmacology
  • Zona Glomerulosa / metabolism*

Substances

  • Calcium Channel Blockers
  • Calcium Channels
  • Dopamine Agonists
  • Dopamine Antagonists
  • Enzyme Inhibitors
  • Flavonoids
  • Receptors, Dopamine D1
  • Thionucleotides
  • Guanosine Diphosphate
  • Guanosine 5'-O-(3-Thiotriphosphate)
  • guanosine 5'-O-(2-thiodiphosphate)
  • Cyclic AMP
  • Calcium-Calmodulin-Dependent Protein Kinases
  • GTP-Binding Protein alpha Subunits, Gs
  • Adenylyl Cyclases
  • adenylyl cyclase 2
  • 2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one
  • Dopamine