Paramecium primaurelia is a unicellular eukaryote that moves in freshwater by ciliary beating and responds to environmental stimuli by altering motile behaviour. The movements of the cilia are controlled by the electrical changes of the cell membrane: when the intraciliary Ca2+ concentration associated with plasma membrane depolarization increases, the ciliary beating reverses its direction, and consequently the swimming direction changes. The ciliary reversal duration is correlated with the amount of Ca2+ influx. Here we evaluated the effects due to the activation or blockade of NMDA receptors on swimming behaviour in Paramecium. Paramecia normally swim forward drawing almost linear tracks. We observed that the simultaneous administration of NMDA and glycine induced a partial ciliary reversal (PaCR) leading to a continuous spiral-like swim. Furthermore, the duration of continuous ciliary reversal (CCR), triggered by high external KCl concentrations, was longer in NMDA/glycine treated cells. NMDA action required the presence of Ca2+, as the normal forward swimming was restored when the ion was omitted from the extracellular milieu. The PaCR and the enhancement of CCR duration significantly decreased when the antagonists of the glutamate site D-AP5 or CGS19755, the NMDA channel blocker MK-801, or the glycine site antagonist DCKA were added. The action of NMDA/glycine was also abolished by Zn2+ or ifenprodil, the GluN2A and the GluN2B NMDA-containing subunit blockers, respectively. Searches of the Paramecium genome database currently available indicate that the NMDA-like receptor with ligand binding characteristics of an NMDA receptor-like complex, purified from rat brain synaptic membranes and found in some metazoan genome, is also present in Paramecium. These results provide evidence that functional NMDA receptors similar to those typical of mammalian neuronal cells are present in the single-celled organism Paramecium and thus suggest that the glutamatergic NMDA system is a phylogenetically old behaviour-controlling mechanism.

Pharmacological characterization of N-methyl-d-aspartic acid (NMDA)-like receptors in the single-celled organism Paramecium primaurelia

RAMOINO, PAOLA;CANDIANI, SIMONA;PITTALUGA, ANNA MARIA;USAI, CESARE;GALLUS, LORENZO;FERRANDO, SARA;MILANESE, MARCO;BONANNO, GIAMBATTISTA
2014-01-01

Abstract

Paramecium primaurelia is a unicellular eukaryote that moves in freshwater by ciliary beating and responds to environmental stimuli by altering motile behaviour. The movements of the cilia are controlled by the electrical changes of the cell membrane: when the intraciliary Ca2+ concentration associated with plasma membrane depolarization increases, the ciliary beating reverses its direction, and consequently the swimming direction changes. The ciliary reversal duration is correlated with the amount of Ca2+ influx. Here we evaluated the effects due to the activation or blockade of NMDA receptors on swimming behaviour in Paramecium. Paramecia normally swim forward drawing almost linear tracks. We observed that the simultaneous administration of NMDA and glycine induced a partial ciliary reversal (PaCR) leading to a continuous spiral-like swim. Furthermore, the duration of continuous ciliary reversal (CCR), triggered by high external KCl concentrations, was longer in NMDA/glycine treated cells. NMDA action required the presence of Ca2+, as the normal forward swimming was restored when the ion was omitted from the extracellular milieu. The PaCR and the enhancement of CCR duration significantly decreased when the antagonists of the glutamate site D-AP5 or CGS19755, the NMDA channel blocker MK-801, or the glycine site antagonist DCKA were added. The action of NMDA/glycine was also abolished by Zn2+ or ifenprodil, the GluN2A and the GluN2B NMDA-containing subunit blockers, respectively. Searches of the Paramecium genome database currently available indicate that the NMDA-like receptor with ligand binding characteristics of an NMDA receptor-like complex, purified from rat brain synaptic membranes and found in some metazoan genome, is also present in Paramecium. These results provide evidence that functional NMDA receptors similar to those typical of mammalian neuronal cells are present in the single-celled organism Paramecium and thus suggest that the glutamatergic NMDA system is a phylogenetically old behaviour-controlling mechanism.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11567/632192
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