Revisiting the postulated "unitary glutamate receptor": electrophysiological and pharmacological analysis in two heterologous expression systems fails to detect evidence for its existence.
Several years ago evidence for a so-called "unitary glutamate receptor" was published. This unique type of glutamate receptor was reported to be activated by the traditional agonists of all three major glutamate receptor subfamilies [i.e., alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA), kainate, and N-methyl-d-aspartate (NMDA)] in a glycine-dependent as well as magnesium-blockable manner and was reported to consist of an NR1 subunit coexpressed with the kainate binding protein (KBP) from Xenopus laevis, XenU1. To re-examine the existence of such a receptor, we cloned two splice variants of the X. laevis NMDA receptor subunit NR1, XenNR1-4a and XenNR1-4b, and expressed them in X. laevis oocytes as well as in human embryonic kidney (HEK) 293 cells, either alone or with the X. laevis KBP subunit XenU1. In addition, we coexpressed XenU1 separately with all eight splice variants of the rat NR1 subunit. In no case did we see evidence of a unitary glutamate receptor pharmacology. In HEK293 cells, we did not get receptor response unless an NR2 subunit was coexpressed. In X. laevis oocytes, we did observe responses to glutamate/glycine as well as small responses to glycine alone, but these were independent of coexpressed XenU1. Neither AMPA nor kainate ever elicited significant responses. Because we verified that XenU1 is expressed and inserted into the plasma membrane of HEK293 cells, we conclude that XenU1 and NR1 do not form the postulated unitary glutamate receptor. Furthermore, successful amplification of a fragment of a X. laevis NR2 subunit indicates that X. laevis uses NR2 subunits and not XenU1 to form heteromeric complexes with NR1.
Schmidt
Carsten
C
Department of Biochemistry I-Receptor Biochemistry, Ruhr University Bochum, Universitaetsstrasse 150, Building NC, Level 6, Room 170, D-44789 Bochum, Germany.
Werner
Markus
M
Hollmann
Michael
M
D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002460:Cell Line; D017931:DNA Primers; D018076:DNA, Complementary; D005260:Female; D005456:Fluorescent Dyes; D006801:Humans; D008969:Molecular Sequence Data; D016296:Mutagenesis; D012326:RNA Splicing; D051381:Rats; D017470:Receptors, Glutamate / Q000145:classification / Q000187:drug effects* / Q000235:genetics / Q000502:physiology*; D014982:Xenopus laevis
D017931:DNA Primers; D018076:DNA, Complementary; D005456:Fluorescent Dyes; D017470:Receptors, Glutamate