The Journal of Neuroscience, June 29, 2005, ():
Reelin, Very-Low-Density Lipoprotein Receptor, and Apolipoprotein E Receptor 2 Control Somatic NMDA Receptor Composition during Hippocampal Maturation In Vitro
J. Neurosci. Sinagra et al.
25: 6127
Supplemental data
Files in this Data Supplement:
- supplemental material
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Supplementary Figure 1. Maturational change of SynCAM expression.
Bar graph showing an increase in total cellular SynCAM protein expression between 6 DIV (100.0±31.3; n=6) and 10 DIV (159.4±26.5; n=6; P<0.01 paired t-test). The level of SynCAM expression was not perturbated at 10 DIV after chronic treatment with –reelin as- (161.6±24.2; n=6) or the control mismatch –reelin m- (172.6±17.4; n=6; NS paired t-test).
- supplemental material
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Supplementary Figure 2. Acute rReelin does not modulate NMDA-evoked currents in 6 or 10 DIV neurons.
A, Representative traces of whole-cell NMDA-evoked currents recorded in a 10 DIV neuron and evoked in the presence of NMDA (left trace) and in the presence of NMDA+rReelin (right trace; calibration : 400pA, 1sec) after 4 min of rReelin application (1µg/ml)
B-C, Average±sem of currents evoked by NMDA+rReelin expressed as the percentage of control NMDA currents evoked by NMDA only in 6 DIV (n=5 cells, B) or 10 DIV (n=5 cells, C) cultures. On each cell, NMDA+rReelin-induced currents were evoked after perfusion of rReelin for various durations (from 30 sec to 7 min). Each bar represent the average±sem calculated after different durations of rReelin bath application.
- supplemental material
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Supplementary Figure 3. Representative western blots using NR2A or NR2B antibodies.
A, B, To control specificity of the NR2A (A) and NR2B (B) antibodies western blots were performed using lysates of COS-7 cells tranfected with either NR2A (lane: 2A) or NR2B (lane: 2B). The other lanes correspond to non transfected COS-7 cells (lane: NT), lysate from either 6 DIV hippocampal cells culture (lane: 6) or whole hippocampi from adult Swiss mice (lane: H).
A, A band with a MW estimated around 190 kDa was recognized by the NR2A antibody (lane: 2A) whereas no signal was detected in lanes NT or 2B. Bands corresponding to MW of approximately 190 kDa were detected in extracts from cell culture or whole hippocampi.
B, The anti-NR2B antibody recognized a band with a MW estimated around 180 kDa in the lane 2B. Bands with similar MW were detected in extracts from cell culture or whole hippocampi, no signal was detected inlanes NT or 2A.
