Figure 9

Transient associations of Nlg and NMDA receptor transport packets (NRTPs) and the effects of Nlg patching on NRTP mobility. (A) Time-lapse imaging of 4 d.i.v. neurons cotransfected with NR1-DsRed (red) and CFP-Nlg1 (green) demonstrates that NRTPs (arrowheads) can move between stable Nlg clusters. Grey arrowheads indicate original location of cluster. Clusters that were immobile during the imaging period are indicated by an asterisk in the top panel. Time is in minutes and seconds. Scale bar, 5 μm. (B) Alternatively, CFP-Nlg1 can move to a stable NR1-DsRed cluster and then the two can move together (arrowheads). Clusters that were immobile during the imaging period are indicated by an asterisk in the top panel. Scale bar, 5 μm. (C) Time-lapse imaging of neurons co-transfected with CFP-Nlg1 (green) and NR1-DsRed (red) after patching with β-NRX-Fc for at least 1 hour. Over 65% of NR1-DsRed was colocalized with CFP-Nlg1 (white arrowheads). One cluster of NR1-DsRed alone is highlighted with a yellow arrowhead. Scale bar, 10 μm. (D) The percentage of total NR1-DsRed clusters that were mobile was significantly decreased after patching with β-NRX-Fc (numbers of neurons analyzed are indicated above bars). Error bars are standard error.