title: Pharmacological characterization of NMDA-like receptors in the single-celled organism Paramecium primaurelia.
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 Ca(2+) 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 Ca(2+) influx. Here, we evaluated the effects due to the activation or blockade of N-methyl-d-aspartic acid (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 Ca(2+), 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 was added. The action of NMDA+glycine was also abolished by Zn(2+) 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 genomes, 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.
authors.affiliation: Department of Earth, Environment and Life Sciences (DISTAV), University of Genoa, Corso Europa 26, 16132 Genoa, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Ramoino
authors.forename: Simona
authors.initials: S
authors.lastname: Candiani
authors.forename: Anna Maria
authors.initials: AM
authors.lastname: Pittaluga
authors.forename: Cesare
authors.initials: C
authors.lastname: Usai
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Gallus
authors.forename: Sara
authors.initials: S
authors.lastname: Ferrando
authors.forename: Marco
authors.initials: M
authors.lastname: Milanese
authors.forename: Marco
authors.initials: M
authors.lastname: Faimali
authors.forename: Giambattista
authors.initials: G
authors.lastname: Bonanno
mesh_terms: D002118:Calcium / Q000378:metabolism; D005998:Glycine / Q000378:metabolism; D016202:N-Methylaspartate / Q000378:metabolism; D010247:Paramecium / Q000187:drug effects / Q000502:physiology*; D015800:Protozoan Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D016194:Receptors, N-Methyl-D-Aspartate / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D013550:Swimming
chemical_list: D015800:Protozoan Proteins; D016194:Receptors, N-Methyl-D-Aspartate; D016202:N-Methylaspartate; D002118:Calcium; D005998:Glycine
keywords: Ciliated protozoa; Glutamatergic NMDA system; NMDA receptor pharmacology; Swimming behaviour