Results: 4

1.
FIGURE 4.

FIGURE 4. From: Dopamine D4 Receptors Regulate GABAA Receptor Trafficking via an Actin/Cofilin/Myosin-dependent Mechanism.

The actin motor protein, myosin, is involved in D4 reduction of GABAAR currents. A, plot of normalized peak GABAAR current as a function of time and PD168077 (30 μm) application in neurons dialyzed with or without the myosin peptide (25 μm). Inset, representative current traces (at time points denoted by #). Scale bar, 200 pA, 1 s. B, cumulative data (mean ± S.E.) showing the percentage of reduction of GABAAR current by PD168077 in a sample of neurons dialyzed with or without the myosin peptide. *, p < 0.005, ANOVA. C, cumulative plots of the distribution of mIPSC amplitudes before (control) and after PD168077 (30 μm) application in cultured PFC neurons dialyzed with or without the myosin peptide (25 μm). Inset, representative mIPSC traces. Scale bar, 50 pA, 1 s. D, cumulative data (mean ± S.E.) showing the percentage of reduction of mIPSC amplitudes by PD168077 in the absence and presence of the myosin peptide. E, plot of normalized peak evoked IPSC as a function of time and PD168077 application in PFC neurons dialyzed with the myosin peptide (25 μm) or a scrambled control peptide (25 μm). F, cumulative data (means ± S.E.) showing the percentage of reduction of evoked IPSC amplitude by PD168077 in the absence and presence of the myosin peptide. *, p < 0.005, ANOVA. PD, PD168077; ctl, control; pep., peptide.

Nicholas M. Graziane, et al. J Biol Chem. 2009 March 27;284(13):8329-8336.
2.
FIGURE 2.

FIGURE 2. From: Dopamine D4 Receptors Regulate GABAA Receptor Trafficking via an Actin/Cofilin/Myosin-dependent Mechanism.

D4 reduction of GABAAR currents is through an actin-dependent mechanism. A, plot of normalized peak GABAAR current as a function of time and PD168077 (30 μm) application in neurons dialyzed with or without the dynamin inhibitory peptide (50 μm). Inset, representative current traces (at time points denoted by #). Scale bar, 500 pA, 1 s. B, cumulative data (means ± S.E.) showing the percentage of reduction of GABAAR current by PD168077 in a sample of neurons in the absence (control) or presence of the dynamin inhibitory peptide. C, plot of normalized peak GABAAR current as a function of time and PD168077 application in neurons dialyzed with the actin stabilizer phalloidin (12.5 μm). Inset, representative current traces (at time points denoted by #). Scale bar, 500 pA, 1 s. D, cumulative data (mean ± S.E.) showing the percent reduction of GABAAR current by PD168077 in the absence or presence of phalloidin in a sample of neurons tested. *, p < 0.005, ANOVA. E, plot of normalized peak GABAAR current as a function of time and PD168077 application in neurons dialyzed with the actin depolymerizer latrunculin B (5 μm) or perfused with the microtubule depolymerizer colchicine (30 μm). F, cumulative data (mean ± S.E.) showing the percent reduction of GABAAR current by PD168077 in the absence or presence of latrunculin B or colchicine in a sample of neurons tested. *, p < 0.005, ANOVA. G, plot of evoked IPSC as a function of time and PD168077 application in a PFC slice perfused with latrunculin B. Inset, representative IPSC traces (at time points denoted by #). Scale bar, 50 pA, 20 ms. ctl, control; PD, PD168077; Dyn. inh. pep., dynamin inhibitory peptide; Lat, latrunculin B; Col, colchicine.

Nicholas M. Graziane, et al. J Biol Chem. 2009 March 27;284(13):8329-8336.
3.
FIGURE 3.

FIGURE 3. From: Dopamine D4 Receptors Regulate GABAA Receptor Trafficking via an Actin/Cofilin/Myosin-dependent Mechanism.

The actin depolymerizing factor, cofilin, is involved in D4 reduction of GABAAR currents. A, left panel, Western blots of p-cofilin and actin in cultured PFC neurons incubated with PD168077 (30 μm, 10 min) in the absence or presence of okadaic acid (1 μm, 40 min pretreatment). Right panel, quantification showing the normalized level of p-cofilin with different treatments. *, p < 0.005, ANOVA. B, plot of evoked IPSC as a function of time and PD168077 application in neurons dialyzed with the Ser3-phosphorylated cofilin peptide p-cof[1–16] (100 μm) or nonphosphorylated cofilin peptide cof[1–16] (100 μm). C, plot of normalized peak GABAAR current as a function of time and PD168077 application in neurons dialyzed with p-cof[1–16] or cof[1–16]. Inset, representative current traces (at time points denoted by #). Scale bar, 500 pA, 1 s. D, cumulative data (means ± S.E.) from acutely dissociated PFC neurons showing the percent reduction of GABAAR current by PD168077 in a sample of neurons dialyzed with different peptides. *, p < 0.005, ANOVA. OA, okadaic acid; PD, PD168077; ctl, control; pep., peptide; scram., scrambled.

Nicholas M. Graziane, et al. J Biol Chem. 2009 March 27;284(13):8329-8336.
4.
FIGURE 1.

FIGURE 1. From: Dopamine D4 Receptors Regulate GABAA Receptor Trafficking via an Actin/Cofilin/Myosin-dependent Mechanism.

D4 receptors reduce GABAAR currents and surface expression in PFC pyramidal neurons. A, plot of normalized peak GABA (50 μm)-evoked current as a function of time in the absence and presence of D4 agonist (PD168077, 30 μm) application in dissociated PFC pyramidal neurons. Inset, representative current traces (at time points denoted by #). Scale bar, 500 pA, 1 s. B, plot of normalized evoked IPSC as a function of time and agonist application in PFC slices. Inset, representative IPSC traces (at time points denoted by #). Scale bar, 25 pA, 20 ms. C, cumulative plot of the distribution of mIPSC amplitudes before (control) and after PD168077 application in a cultured PFC pyramidal neuron. Inset, representative mIPSC traces. Scale bar, 50 pA, 2 s. D, immunocytochemical images of surface GABAAR β2/3 subunits and MAP2 staining in PFC cultures either untreated (control) or treated with PD168077 (30 μm, 10 min). E, quantitative analysis of surface GABAAR β2/3 clusters along the dendrites (density, size, and normalized intensity) in control versus PD168077-treated neurons. *, p < 0.05, ANOVA. ctl or con, control; PD,. PD168077.

Nicholas M. Graziane, et al. J Biol Chem. 2009 March 27;284(13):8329-8336.

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