Role of calcium in neurotensin-evoked enhancement in firing in mesencephalic dopamine neurons.
Neurotensin (NT) increases neurotransmission within the mesolimbic dopamine system by enhancing the firing rate of dopaminergic (DAergic) neurons and by acting at the nerve terminal level. The signal transduction pathways involved in these effects have not been characterized, but NT receptors are coupled to the phospholipase C pathway and Ca(2+) mobilization. However, an enhancement of intracellular Ca(2+) concentration ([Ca(2+)](i)) evoked by NT in DAergic neurons has yet to be demonstrated. Furthermore, the hypothesis that the excitatory effects of NT in DAergic neurons are Ca(2+) dependent is currently untested. In whole-cell recording experiments, DAergic neurons in culture were identified by their selective ability to express a cell-specific green fluorescent protein reporter construct. These experiments confirmed that NT increases firing rate in cultured DAergic neurons. This effect was Ca(2+) dependent because it was blocked by intracellular dialysis with BAPTA. Using Ca(2+) imaging, we showed that NT caused a rapid increase in [Ca(2+)](i) in DAergic neurons. Most of the Ca(2+) originated from the extracellular medium. NT-induced excitation and Ca(2+) influx were blocked by SR48692, an antagonist of the type 1 NT receptor. Blocking IP(3) receptors using heparin prevented the excitatory effect of NT. Moreover, Zn(2+) and SKF96365 both blocked the excitatory effect of NT, suggesting that nonselective cationic conductances are involved. Finally, although NT can also induce a rise in [Ca(2+)](i) in astrocytes, we find that NT-evoked excitation of DAergic neurons can occur independently of astrocyte activation.
St-Gelais
Fannie
F
Département de Pharmacologie, Centre de Recherche en Sciences Neurologiques, Université de Montréal, Montréal, Québec, Canada H3T IJ4.
Legault
Mark
M
Bourque
Marie-Josée
MJ
Rompré
Pierre-Paul
PP
Trudeau
Louis-Eric
LE
D000818:Animals; D001253:Astrocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D002118:Calcium / Q000502:physiology*; D002121:Calcium Channel Blockers / Q000494:pharmacology; D015220:Calcium Channels; D002478:Cells, Cultured; D002614:Chelating Agents / Q000494:pharmacology; D004298:Dopamine / Q000378:metabolism*; D050483:Dopamine Plasma Membrane Transport Proteins; D004533:Egtazic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology; D017930:Genes, Reporter / Q000235:genetics; D049452:Green Fluorescent Proteins; D006493:Heparin / Q000494:pharmacology; D053496:Inositol 1,4,5-Trisphosphate Receptors; D008164:Luminescent Proteins / Q000235:genetics; D008562:Membrane Glycoproteins; D026901:Membrane Transport Proteins / Q000235:genetics; D008636:Mesencephalon / Q000166:cytology / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000235:genetics; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D009496:Neurotensin / Q000494:pharmacology*; D018408:Patch-Clamp Techniques; D011720:Pyrazoles / Q000494:pharmacology; D011804:Quinolines / Q000494:pharmacology; D051381:Rats; D018160:Receptors, Cytoplasmic and Nuclear / Q000037:antagonists & inhibitors; D018028:Receptors, Neurotensin / Q000037:antagonists & inhibitors; D009435:Synaptic Transmission / Q000187:drug effects / Q000502:physiology*; D014162:Transfection
D002121:Calcium Channel Blockers; D015220:Calcium Channels; D002614:Chelating Agents; D050483:Dopamine Plasma Membrane Transport Proteins; D053496:Inositol 1,4,5-Trisphosphate Receptors; D008164:Luminescent Proteins; D008562:Membrane Glycoproteins; D026901:Membrane Transport Proteins; D009419:Nerve Tissue Proteins; D011720:Pyrazoles; D011804:Quinolines; D018160:Receptors, Cytoplasmic and Nuclear; D018028:Receptors, Neurotensin; C114036:neurotensin type 1 receptor; C079087:SR 48692; D049452:Green Fluorescent Proteins; D009496:Neurotensin; D004533:Egtazic Acid; D006493:Heparin; C025603:1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid; D002118:Calcium; D004298:Dopamine