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The glutamate receptor originally called GluR-KI (15) was shown to be activated by both KA and AMPA (17-19). Thus, the existence of separate KA and AMPA receptors is uncertain, and we therefore refer to GluR-K1 as a KA-AMPA receptor. We also changed the original name GluR-K1 to the pharmacologically neutral name GluR1 (17).
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2, NMG for osmotic balance, 10 mM Hepes, pH adjusted to 7.2 with HCl. We prepared cRNA transcripts and performed oocyte injections as described (15). Recordings were made 4 to 10 days after cRNA injection under two-electrode voltage damp at a holding potential of - 70 mV (except for I-V curves) with a DAGAN 8500 amplifier. Voltage electrodes had a resistance of 4 to 8 megohms and were filled with 3 M KCl; current electrodes had a resistance of∼1 megohm and were filled with 0.25 M CsCl, 0.25 M CsF, 100 mM EGTA, pH 7.2. All drugs were pulse-applied (1.6 ml) by superfusion (4 ml/min) in a 300-μl chamber; peak currents were recorded.
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2+-Ringer strongly depended on the relative proportion of the two cRNAs that were injected. If GluR3 and GluR2 cRNAs were injected at a molar ratio of 1:1, some inward current and a slightly inwardly rectifying I-V curve were observed (Fig. 3B). However, when GluR2 was injected in tenfold molar excess, no inward current was seen and the I-V curve was outwardly rectifying (Fig. 3B). This is interpreted as follows: when the two cRNAs are injected at a 1:1 ratio, homooligomeric channels and heterooligomeric channels are formed, with both types of channels contributing some current; with GluR2 in excess, formation of homooligomeric GluR3 channels presumably is reduced, and the response is largely due to the heterooligomeric channels. Because GluR2 by itself gives only very weak responses in oocytes (17,19), its tenfold excess (which presumably results in a large number of homooligomeric GluR2 channels) does not contribute significantly to the current measured and thus does not obscure the analysis of heterooligomeric channel properties. In contrast to the subunit combinations involving GluR2, the GluR1 and GluR3 channels do not seem to interact in the oocyte membrane, at least not in such a way as to form heterooligomeric channels with permeability properties different from those of the respective homooligomeric channels.
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We thank J. Boulter, R. Dingledine, T. Hughes, R. Hume, R. Papke, and C. Stevens for helpful discussions and critical reading of the manuscript. Supported by a fellowship from the Deutsche Forschungsgemeinschaft (M.H.) and grants from the NIH (NS 28709-01), the Muscular Dystrophy Association, me Human Frontier Science Program, and the Fritz B. Burns foundation (S.H.).
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