Journal:

Article Title: POSITIVE AMPA RECEPTOR MODULATION RAPIDLY STIMULATES BDNF RELEASE AND INCREASES DENDRITIC mRNA TRANSLATION

doi: 10.1523/JNEUROSCI.6078-08.2009

Figure Lengend Snippet: a. Cultured cortical neurons (DIV14) were preincubated with TrkB-Fc (0.5 μg/mL, 1h) before being treated with CX614 (10 μM, 1 h). At the end of treatment, cells were processed for immunoblotting with antibodies against actin and total and phosphorylated mTOR. Immunoblots were quantified and ratios of p-mTOR over total unphosphorylated mTOR were calculated; data were then expressed as fold of control and are means ± S.E.M. from 6 independent experiments. * p < 0.001 as compared to control; † p < 0.05 as compared to CX614; # not significantly different from control (ANOVA followed by Tukey’s post-test analysis).

Article Snippet: Primary neuronal cultures and acute hippocampal slices were treated with the following blockers and inhibitors for 10 and 30 min, respectively, before being treated with CX614 or BDNF; these inhibitors and blockers were used at the following concentrations: (2 R )-amino-5-phosphonovaleric acid (APV) (Tocris; 25 μM in cultured neurons), 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) (Tocris; 50 μM in slices; 25 μM in cultured neurons), (9S,10R,12R)-2,3,9,10,11,12-Hexahydro-10-hydroxy-9-methyl-1-oxo-9,12-epoxy-1Hdiindolo[1,2,3-fg:3′,2′,1′-kl]pyrrolo[3,4-i][1,6]benzodiazocine-10-carboxylic acid methyl ester (K252a) (Tocris; 0.5 μM), rapamycin (Calbiochem; 1 μM), actinomycin D (Calbiochem; 10 μM), nifedipine (Sigma; 10 μM), ryanodine (Calbiochem; 100 μM), EGTA (Sigma; 5 mM).

Techniques: Cell Culture, Western Blot, Control

Journal:

Article Title: POSITIVE AMPA RECEPTOR MODULATION RAPIDLY STIMULATES BDNF RELEASE AND INCREASES DENDRITIC mRNA TRANSLATION

doi: 10.1523/JNEUROSCI.6078-08.2009

Figure Lengend Snippet: a. Acute hippocampal slices from adult male rats incubated with vehicle (Control) or CX614 (50 μM) for 1 h were immediately processed for immunohistochemistry with antibodies against phospho-mTOR (left panels) or phospho-4EBP1 (right panels). Note the clear increase in immunoreactivity in stratum radiatum of CA1 and in stratum granulosum of the dentate gyrus (DG) in CX614-treated slices compared to control. Scale bar: 20 μm.

Article Snippet: Primary neuronal cultures and acute hippocampal slices were treated with the following blockers and inhibitors for 10 and 30 min, respectively, before being treated with CX614 or BDNF; these inhibitors and blockers were used at the following concentrations: (2 R )-amino-5-phosphonovaleric acid (APV) (Tocris; 25 μM in cultured neurons), 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) (Tocris; 50 μM in slices; 25 μM in cultured neurons), (9S,10R,12R)-2,3,9,10,11,12-Hexahydro-10-hydroxy-9-methyl-1-oxo-9,12-epoxy-1Hdiindolo[1,2,3-fg:3′,2′,1′-kl]pyrrolo[3,4-i][1,6]benzodiazocine-10-carboxylic acid methyl ester (K252a) (Tocris; 0.5 μM), rapamycin (Calbiochem; 1 μM), actinomycin D (Calbiochem; 10 μM), nifedipine (Sigma; 10 μM), ryanodine (Calbiochem; 100 μM), EGTA (Sigma; 5 mM).

Techniques: Incubation, Control, Immunohistochemistry

Journal:

Article Title: POSITIVE AMPA RECEPTOR MODULATION RAPIDLY STIMULATES BDNF RELEASE AND INCREASES DENDRITIC mRNA TRANSLATION

doi: 10.1523/JNEUROSCI.6078-08.2009

Figure Lengend Snippet: Cultured cortical neurons were incubated with CX614 (10 μM) or BDNF (50 ng/mL) for 1 h in the absence or presence of rapamycin (1 μM). At the end of treatment, cultures were fixed and processed for immunostaining (a) with antibodies against phospho-4EBP1, or lysed and processed for immunoblotting (b) with antibodies against 4EPP1 and phospho-4EBP1. Immunoblots were quantified and ratios of phosphorylated over total 4EBP1 were calculated; data were then expressed as fold of control and are means ± S.E.M. from 6 independent experiments. * p < 0.01, ** p < 0.001 as compared to control; † p < 0.01 as compared to CX614 (ANOVA followed by Tukey’s post-test analysis). Scale bar: 20 μm.

Article Snippet: Primary neuronal cultures and acute hippocampal slices were treated with the following blockers and inhibitors for 10 and 30 min, respectively, before being treated with CX614 or BDNF; these inhibitors and blockers were used at the following concentrations: (2 R )-amino-5-phosphonovaleric acid (APV) (Tocris; 25 μM in cultured neurons), 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) (Tocris; 50 μM in slices; 25 μM in cultured neurons), (9S,10R,12R)-2,3,9,10,11,12-Hexahydro-10-hydroxy-9-methyl-1-oxo-9,12-epoxy-1Hdiindolo[1,2,3-fg:3′,2′,1′-kl]pyrrolo[3,4-i][1,6]benzodiazocine-10-carboxylic acid methyl ester (K252a) (Tocris; 0.5 μM), rapamycin (Calbiochem; 1 μM), actinomycin D (Calbiochem; 10 μM), nifedipine (Sigma; 10 μM), ryanodine (Calbiochem; 100 μM), EGTA (Sigma; 5 mM).

Techniques: Cell Culture, Incubation, Immunostaining, Western Blot, Control

Journal:

Article Title: POSITIVE AMPA RECEPTOR MODULATION RAPIDLY STIMULATES BDNF RELEASE AND INCREASES DENDRITIC mRNA TRANSLATION

doi: 10.1523/JNEUROSCI.6078-08.2009

Figure Lengend Snippet: a. Acute hippocampal slices were treated with CX614 (50 μM) in the absence or presence of CNQX (50 μM) for 1 h. At the end of treatment, tissues were processed for immunoblotting with antibodies against TrkB, phospho-TrkB, BDNF, and actin. Immunoblots were quantified and ratios of phosphorylated over total TrkB were calculated; data were then expressed as fold of control and are means ± S.E.M. from 6 independent experiments. * p < 0.01 as compared to control; † p < 0.01 as compared to CX614 (ANOVA followed by Tukey’s post-test analysis).

Article Snippet: Primary neuronal cultures and acute hippocampal slices were treated with the following blockers and inhibitors for 10 and 30 min, respectively, before being treated with CX614 or BDNF; these inhibitors and blockers were used at the following concentrations: (2 R )-amino-5-phosphonovaleric acid (APV) (Tocris; 25 μM in cultured neurons), 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) (Tocris; 50 μM in slices; 25 μM in cultured neurons), (9S,10R,12R)-2,3,9,10,11,12-Hexahydro-10-hydroxy-9-methyl-1-oxo-9,12-epoxy-1Hdiindolo[1,2,3-fg:3′,2′,1′-kl]pyrrolo[3,4-i][1,6]benzodiazocine-10-carboxylic acid methyl ester (K252a) (Tocris; 0.5 μM), rapamycin (Calbiochem; 1 μM), actinomycin D (Calbiochem; 10 μM), nifedipine (Sigma; 10 μM), ryanodine (Calbiochem; 100 μM), EGTA (Sigma; 5 mM).

Techniques: Western Blot, Control

Journal:

Article Title: POSITIVE AMPA RECEPTOR MODULATION RAPIDLY STIMULATES BDNF RELEASE AND INCREASES DENDRITIC mRNA TRANSLATION

doi: 10.1523/JNEUROSCI.6078-08.2009

Figure Lengend Snippet: a. Cultured cortical neurons (DIV14) were preincubated with TrkB-Fc (0.5 μg/mL, 1 h) and incubated in the absence or presence of CX614 (10 μM, 1 h). At the end of treatment, cells were processed for immunoblotting with antibodies against actin and total and phosphorylated TrkB. Immunoblots were quantified and ratios of p-TrkB over total unphosphorylated TrkB were calculated; data were then expressed as fold of control and are means ± S.E.M. from 6 experiments. * p < 0.001 as compared to control; † p < 0.05 as compared to CX614; # not significantly different from control (ANOVA followed by Tukey’s post-test analysis).

Article Snippet: Primary neuronal cultures and acute hippocampal slices were treated with the following blockers and inhibitors for 10 and 30 min, respectively, before being treated with CX614 or BDNF; these inhibitors and blockers were used at the following concentrations: (2 R )-amino-5-phosphonovaleric acid (APV) (Tocris; 25 μM in cultured neurons), 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) (Tocris; 50 μM in slices; 25 μM in cultured neurons), (9S,10R,12R)-2,3,9,10,11,12-Hexahydro-10-hydroxy-9-methyl-1-oxo-9,12-epoxy-1Hdiindolo[1,2,3-fg:3′,2′,1′-kl]pyrrolo[3,4-i][1,6]benzodiazocine-10-carboxylic acid methyl ester (K252a) (Tocris; 0.5 μM), rapamycin (Calbiochem; 1 μM), actinomycin D (Calbiochem; 10 μM), nifedipine (Sigma; 10 μM), ryanodine (Calbiochem; 100 μM), EGTA (Sigma; 5 mM).

Techniques: Cell Culture, Incubation, Western Blot, Control

Journal:

Article Title: POSITIVE AMPA RECEPTOR MODULATION RAPIDLY STIMULATES BDNF RELEASE AND INCREASES DENDRITIC mRNA TRANSLATION

doi: 10.1523/JNEUROSCI.6078-08.2009

Figure Lengend Snippet: Acute hippocampal slices were treated with CX614 (50 μM) in the absence or presence of Ca++ (a), the absence or presence of ryanodine (100 μM, b) or the absence or presence of nifedipine (10 μM, c) for 1 h. At the end of treatment, tissues were processed for immunoblotting with antibodies against total and phosphorylated TrkB, and actin (as a loading control). Immunoblots were quantified and ratios of phosphorylated over total TrkB were calculated; data were then expressed as percent of control and are means ± S.E.M. from 6 independent experiments. * p < 0.01 as compared to control; † p < 0.01 as compared to CX614 (ANOVA followed by Tukey’s post-test analysis).

Article Snippet: Primary neuronal cultures and acute hippocampal slices were treated with the following blockers and inhibitors for 10 and 30 min, respectively, before being treated with CX614 or BDNF; these inhibitors and blockers were used at the following concentrations: (2 R )-amino-5-phosphonovaleric acid (APV) (Tocris; 25 μM in cultured neurons), 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) (Tocris; 50 μM in slices; 25 μM in cultured neurons), (9S,10R,12R)-2,3,9,10,11,12-Hexahydro-10-hydroxy-9-methyl-1-oxo-9,12-epoxy-1Hdiindolo[1,2,3-fg:3′,2′,1′-kl]pyrrolo[3,4-i][1,6]benzodiazocine-10-carboxylic acid methyl ester (K252a) (Tocris; 0.5 μM), rapamycin (Calbiochem; 1 μM), actinomycin D (Calbiochem; 10 μM), nifedipine (Sigma; 10 μM), ryanodine (Calbiochem; 100 μM), EGTA (Sigma; 5 mM).

Techniques: Western Blot, Control

Journal:

Article Title: POSITIVE AMPA RECEPTOR MODULATION RAPIDLY STIMULATES BDNF RELEASE AND INCREASES DENDRITIC mRNA TRANSLATION

doi: 10.1523/JNEUROSCI.6078-08.2009

Figure Lengend Snippet: Cultured cortical neurons were incubated in the absence or presence of CX614 (10 μM) and K252a (0.5 μM, a), TrkB-Fc (0.5 μg/mL, b), CNQX (50μM, c) or rapamycin (100 μM, d) for 1 h. At the end of treatment, cells were processed for immunoblotting with antibodies against ARC and actin (for loading control). Immunoblots were quantified and corrected ARC levels were calculated; data were then expressed as fold of control and are means ± S.E.M. from 6 independent experiments. * p < 0.01, ** p < 0.001 as compared to control; † p < 0.01 as compared to CX614 (ANOVA followed by Tukey’s post-test analysis).

Article Snippet: Primary neuronal cultures and acute hippocampal slices were treated with the following blockers and inhibitors for 10 and 30 min, respectively, before being treated with CX614 or BDNF; these inhibitors and blockers were used at the following concentrations: (2 R )-amino-5-phosphonovaleric acid (APV) (Tocris; 25 μM in cultured neurons), 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) (Tocris; 50 μM in slices; 25 μM in cultured neurons), (9S,10R,12R)-2,3,9,10,11,12-Hexahydro-10-hydroxy-9-methyl-1-oxo-9,12-epoxy-1Hdiindolo[1,2,3-fg:3′,2′,1′-kl]pyrrolo[3,4-i][1,6]benzodiazocine-10-carboxylic acid methyl ester (K252a) (Tocris; 0.5 μM), rapamycin (Calbiochem; 1 μM), actinomycin D (Calbiochem; 10 μM), nifedipine (Sigma; 10 μM), ryanodine (Calbiochem; 100 μM), EGTA (Sigma; 5 mM).

Techniques: Cell Culture, Incubation, Western Blot, Control

Journal:

Article Title: POSITIVE AMPA RECEPTOR MODULATION RAPIDLY STIMULATES BDNF RELEASE AND INCREASES DENDRITIC mRNA TRANSLATION

doi: 10.1523/JNEUROSCI.6078-08.2009

Figure Lengend Snippet: a. Selected dendrites of cultured hippocampal neurons that were injected with the GFP reporter plasmid at 12 DIV were “photobleached” until fluorescence was almost invisible before being treated with vehicle (Control), BDNF (100 ng/mL), or CX614 (50 μM). Time-lapse images were then taken at 30-second intervals. Scale bar: 5 μm.

Article Snippet: Primary neuronal cultures and acute hippocampal slices were treated with the following blockers and inhibitors for 10 and 30 min, respectively, before being treated with CX614 or BDNF; these inhibitors and blockers were used at the following concentrations: (2 R )-amino-5-phosphonovaleric acid (APV) (Tocris; 25 μM in cultured neurons), 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) (Tocris; 50 μM in slices; 25 μM in cultured neurons), (9S,10R,12R)-2,3,9,10,11,12-Hexahydro-10-hydroxy-9-methyl-1-oxo-9,12-epoxy-1Hdiindolo[1,2,3-fg:3′,2′,1′-kl]pyrrolo[3,4-i][1,6]benzodiazocine-10-carboxylic acid methyl ester (K252a) (Tocris; 0.5 μM), rapamycin (Calbiochem; 1 μM), actinomycin D (Calbiochem; 10 μM), nifedipine (Sigma; 10 μM), ryanodine (Calbiochem; 100 μM), EGTA (Sigma; 5 mM).

Techniques: Cell Culture, Injection, Plasmid Preparation, Fluorescence, Control