The Journal of Neuroscience, May 30, 2007, ():
Mitogen-Activated Protein Kinase Upregulates the Dendritic Translation Machinery in Long-Term Potentiation by Controlling the Mammalian Target of Rapamycin Pathway
J. Neurosci. Tsokas et al.
27: 5885
Supplemental Data
Files in this Data Supplement:
- supplemental material
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Figure S1. Inhibition of ERK does not reduce basal expression of TOP mRNA-encoded proteins or phosphorylation of RSK, PDK1, or Akt. Slices were exposed to 50 µM PD98059 or vehicle for 90 minutes, followed by freezing and excision of area CA1 for subsequent immunoblotting of total eEF1A, eEF2, rpS6, and PABP (light bars), and for phospho-S386 RSK1, phospho-S241 PDK1, and phospho-T308 Akt (dark bars). No differences were detected between drug-treated and vehicle-treated slices for any of the measured proteins. Data are shown as means ± sem of immunoreactivity, normalized to the vehicle-treated controls for each of 5 independent experiments.
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Figure S2. HFS does not affect the expression of signaling components. Slices were frozen 30 min after the delivery of HFS or sham stimulation (see Materials and Methods), and immunoblots of CA1 lysates were probed for total RSK (n=5), PDK1 (n=5), mTOR (n=7), and p70S6K (n=5). No significant differences in immunoreactivity were found for any of the proteins. Data are shown as means ± sem of immunoreactivity, normalized to the sham-stimulated controls.
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Figure S3. PI3K activity is required for HFS-induced LTP. Slices were exposed to 20 µM LY294002 or vehicle (0.1% DMSO) (indicated by bar) for 30 min before the delivery of HFS or control stimulation. Basal synaptic efficiency was depressed by LY294002 (n = 5), in agreement with Opazo et al (2002) but not with Sanna et al. (2002). In drug-treated slices stimulated with HFS, LTP over the last 5 min of recording (23.8 ± 1.1%, measured against the drug-treated controls at the same time points; n = 6) was significantly smaller (p < .01) than LTP in vehicle-treated slices (+45.2 ± 0.4%, measured against the last 5 min of baseline; n = 5).
