Group I mGluRs coupled to G proteins are regulated by tyrosine kinase in dopamine neurons of the rat midbrain

Alessandro Tozzi, Ezia Guatteo, Luigi Caputi, Giorgio Bernardi, Nicola B. Mercuri

Research output: Contribution to journalArticlepeer-review

Abstract

Metabotropic glutamate receptors (mGluRs) modulate neuronal function via different transduction mechanisms that are either dependent or independent on G-protein function. Here we investigated, using whole cell patch-clamp recordings in combination with fluorimetric measurements of intracellular calcium concentration ([Ca 2+] i), the metabolic pathways involved in the responses induced by group I mGluRs in dopamine neurons of the rat midbrain. The inward current and the [Ca 2+] i increase caused by the group I mGluR agonist (S)-3,5-dihydroxyphenylglycine (DHPG, 100 μM) were permanently activated and subsequently abolished in cells loaded with the nonhydrolizable GTP-analogue GTP-γ-S (600 μM). In addition, when GDP-β-S (600 μM) was dialyzed into the cells to produce the blockade of the G proteins, the DHPG-dependent responses were reduced. When the tissue was bathed with the phospholipase C inhibitor 1-[6[[(17β)-3-methoxyestra-1,3,5(10)-trien-17-yl] amino]exyl]-1H-pyrrole-2,5-dione (10 μM), the DHPG-induced calcium transients slightly diminished but the associated inward currents were not affected. Interestingly, a substantial depression of the DHPG-induced inward current and transient increase of [Ca 2+] i was caused by the protein tyrosine kinase inhibitors tyrphostin B52 (40 μM) and 4′,5,7-trihydroxyisoflavone (genistein; 40 μM), whereas genistein's inactive analogue 4′,5,7-trihydroxyisoflavone-7-glucoside (40 μM) was ineffective. The blockade of the Src family of tyrosine kinase by 4-amino-5-(4-methylphenyl)-7-(t-butyl)pyrazolo[3,4-d]-pyrimidine (20 μM), mitogen-activated protein kinase by 2′-amino-3′ methoxyflavone (50 μM), and protein kinase C by staurosporine (1 μM) had no effect on the cellular responses caused by DHPG. The mGluR5-selective antagonist 2-methyl-6-(phenylethynyl)-pyridine (10-100 μM) did not affect the actions of DHPG. Thus our results indicate that the responses, mainly mediated by mGluRs1 in dopamine neurons, are activated by intracellular mechanisms coupled to G proteins and regulated by tyrosine kinases.

Original languageEnglish
Pages (from-to)2490-2497
Number of pages8
JournalJournal of Neurophysiology
Volume85
Issue number6
Publication statusPublished - 2001

ASJC Scopus subject areas

  • Physiology
  • Neuroscience(all)

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