The Journal of Neuroscience, August 15, 2007, ():
TrpC3/C7 and Slo2.1 Are Molecular Targets for Metabotropic Glutamate Receptor Signaling in Rat Striatal Cholinergic Interneurons
J. Neurosci. Berg et al.
27: 8845
Supplemental Data
Files in this Data Supplement:
- supplemental material
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Supplemental Figure 1. Activation kinetics of Slo2.1 and the Cl- activated and mGluR1/5 receptor-sensitive current in striatal interneurons. Averaged currents evoked by voltage step commands (from -125 mV to -5 mV) in HEK293T cells expressing Slo2.1 (n=5) and in striatal interneurons recorded with a KCl-based pipette solution (n=5). In interneurons, currents activated by voltage steps in the presence of 3,5-DHPG were subtracted from those obtained immediately after run-up. Note the essentially instantaneous current kinetics of both Slo2.1 and the native Cl- activated and DHPG-sensitive current.
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Supplemental Figure 2. Slo2.1 (Slick) and Slo2.2 (Slack) are inhibited by activation of G?q-coupled TRH receptor. HEK293 cells stably expressing the thyrotropin-releasing hormone (TRH) receptor E2 cells (Kim et al., 1994) obtained from G. Milligan (University of Glasgow, Scotland, UK) were transfected with either GFP-Slick fusion construct or Slack-pcDNA3. For recordings of Slack currents, the pipette solution contained (mM): 110 KCl, 20 NaCl, 10 HEPES K, 5 EGTA, 3 ATP, 0.3 GTP. Under whole-cell voltage clamp, cells were held at -60 mV and depolarizing ramps (from -130 mV to +20 mV) applied every five seconds under control conditions and during exposure to TRH (0.2 µM); slope conductance was measured between -80 and -35 mV. A. Representative time series illustrate inhibition of rSlo2.1 and rSlo2.2 by TRH receptor activation. B. The magnitude of Slo2.1 and Slo2.2 inhibition by TRH (as % of control conductance).
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Supplemental Figure 3. High intracellular Cl- does not preclude mGluR1 activation of TrpC3 in HEK293T cells. HEK293T cells were transfected with TrpC3 and mGluR1 and recorded under whole cell voltage clamp with the same KCl-based pipette solution used to record from striatal interneurons. Currents were evoked by ramp voltage commands under control conditions and in the presence of 3,5-DHPG (upper); even in the presence of high Cl-i, the agonist activated a current with a doubly rectifying profile expected of TrpC3 (lower).
