Journal: Annals of Translational Medicine

Article Title: The recycling of AMPA receptors/GABAa receptors is related to neuronal excitation/inhibition imbalance and may be regulated by KIF5A

doi: 10.21037/atm-22-4337

Figure Lengend Snippet: Expression of KIF5A, GluR2 and beta 2+3 subunits of gamma aminobutyric acid receptors (Gabrb2+3) in an in vivo model of seizures. (A) EEG results: There was no epileptic discharge in the Ctl group after the injection of saline; however, epileptic discharge was observed in the Sez group after the injection of PTZ. (B) Total protein expression: the gray level of the total protein expression bands was normalized with GAPDH, and the total protein expression levels of KIF5A, GluR2 and Gabrb2+3 in the hippocampus of the Sez group did not change significantly (n=6 in each group, vs. Ctl, P>0.05). (C) Surface protein expression: The gray level of the total protein expression bands was normalized with Sodium/potassium-transporting ATPase subunit alpha-1 (ATP1A1). In the Sez group, the expression of GluR2 on the surface increased significantly to 181.74%±14.44% ( vs. Ctl, # , P<0.01). Conversely, the expression of GluR2 on the surface decreased to 19.62%±8.01% ( vs. Ctl, # , P<0.01). KIF5A, kinesin superfamily proteins 5A; GluR2, glutamate receptors subunit-2; Ctl, control; Sez, seizures; PTZ, pentylenetetrazol.

Article Snippet: The primary antibodies used include anti-KIF5A antibody 1:1,000 (SANTA, sc-376452), anti-GluR2 antibody 1:1,000 (BOSTER, PB9205), and anti-GABAaR β2+3 antibody 1:500 (Bioss, bs-12066R).

Techniques: Expressing, In Vivo, Injection

Journal: Annals of Translational Medicine

Article Title: The recycling of AMPA receptors/GABAa receptors is related to neuronal excitation/inhibition imbalance and may be regulated by KIF5A

doi: 10.21037/atm-22-4337

Figure Lengend Snippet: Receptor recycling assay (IF). The recycling ratio of GluR2 was 0.30±0.05 in the Ctl group and 0.60±0.07 in the Mg 2+ -free group (7 cells per group, vs. Ctl, # , P<0.01), and the recycling ratio of Gabrb2+3 was 0.49±0.04 in Ctl group and 0.32±0.05 in the Mg 2+ -free group (7 cells per group, vs. Ctl, # , P<0.01) (×600). Scale Bar: 50 µm. IF, immunofluorescence; Ctl, control.

Article Snippet: The primary antibodies used include anti-KIF5A antibody 1:1,000 (SANTA, sc-376452), anti-GluR2 antibody 1:1,000 (BOSTER, PB9205), and anti-GABAaR β2+3 antibody 1:500 (Bioss, bs-12066R).

Techniques: Immunofluorescence

Journal: Annals of Translational Medicine

Article Title: The recycling of AMPA receptors/GABAa receptors is related to neuronal excitation/inhibition imbalance and may be regulated by KIF5A

doi: 10.21037/atm-22-4337

Figure Lengend Snippet: Interaction between KIF5A and GluR2 and Gabrb2+3 in the seizure model. (A) Co-ip results: The protein bands showed the co-ip levels of KIF5A, GluR2, and Gabrb2+3, and normalized with the protein levels of KIF5A. The GluR2 level of KIF5A pull-down in the hippocampi of the rats in the Sez group increased to 130.42%±53.24% (n=6 per, vs. Ctl, *, P<0.05). However, the Gabrb2+3 level of KIF5A decreased to 50.86%±5.33% in the Sez group (n=6 per group, vs. Ctl, # , P<0.01). (B) IF results: the Pearson’s correlation coefficients (PCC) of KIF5A/GluR2 was 0.40±0.19 in the Ctl group and 0.87±0.11 in the Mg 2+ -free solution group (n=6 per group, vs. Ctl, # , P<0.01) (×400). Scale Bar: 100 µm. (C) IF results: the PCC of KIF5A/Gabrb2+3 was 0.97±0.02 in the Ctl group and 0.32±0.11 in the Mg 2+ -free solution group (n=6 per group, vs. Ctl, # , P<0.01) (×400). Scale Bar: 100 µm. IF, immunofluorescence.

Article Snippet: The primary antibodies used include anti-KIF5A antibody 1:1,000 (SANTA, sc-376452), anti-GluR2 antibody 1:1,000 (BOSTER, PB9205), and anti-GABAaR β2+3 antibody 1:500 (Bioss, bs-12066R).

Techniques: Co-Immunoprecipitation Assay, Immunofluorescence

Journal: Neuron

Article Title: Regulation of Thalamic and Cortical Network Synchrony by Scn8a

doi: 10.1016/j.neuron.2017.01.031

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Monoclonal mouse anti-GluR2 , Neuromab , Cat# 75-002; RRID: AB_2232661.

Techniques: Plasmid Preparation, Virus, Recombinant, Avidin-Biotin Assay, Software, Imaging

Journal: eNeuro

Article Title: SAP97 Binding Partner CRIPT Promotes Dendrite Growth In Vitro and In Vivo

doi: 10.1523/ENEURO.0175-17.2017

Figure Lengend Snippet: CRIPT knockdown leads to a selective reduction in the abundance of GluA1 and SAP97. Mixed spinal cord cultures were infected with HSV engineered to express a miRNA targeting CRIPT or a scrambled control. Two days later, lysates were prepared and subjected to Western blottings. No more than six independent experiments were performed for the quantitative image analysis. CRIPT knockdown leads to a reduction in GluA1 and SAP97 abundance and no effect on the abundance of GluA2, GluA4, NR1, NR2A, NR2B, or PSD95. Representative images of Western blottings with actin loading controls are shown and quantification of band intensity in the bar graphs below; *significant difference between groups, p < 0.05.

Article Snippet: The following antibodies were used in biochemical assays: immunoprecipitation of SAP97 (Thermo Fisher Scientific, catalog #PA1-741), immunoblotting and immunoprecipitation of CRIPT (Protein Tech Group, catalog #11211-1-AP), immunoprecipitation and immunoblotting of HA-tag (BioLegend, 16B11, catalog #901512), immunoblotting of SAP97 (NeuroMab/Antibodies, catalog #73-030), immunoprecipitation and immunoblotting of the myc-tag (Cell Signaling Technology, 9B11 catalog #2276), immunoblotting GluA1 (NeuroMab/Antibodies, catalog #75-327), immunoblotting GluA2 (NeuroMab/Antibodies, catalog #75-002), immunoblotting GluA4 (Cell Signaling Technology, catalog #8070), immunoblotting NR1 (BD PharMingen, catalog #556308), immunoblotting NR2A (Alomone Labs, catalog #AGC002), immunoblotting NR2B (Alomone Labs, catalog #AGC003), and immunoblotting actin [Cell Signaling Technology, catalog #3700 (mouse) or Sigma-Aldrich, catalog #A2066 (rabbit)].

Techniques: Knockdown, Infection, Control, Western Blot

Journal: Current Research in Toxicology

Article Title: Developmental deltamethrin: Sex-specific hippocampal effects in Sprague Dawley rats

doi: 10.1016/j.crtox.2022.100093

Figure Lengend Snippet: AMPA receptor subunit western blots: (A) GLUR1, (B) GLUR2 protein expression in hippocampus, nucleus accumbens, and striatum. Data are normalized to actin. GLUR1 (∼102 kDa) and GLUR2 (∼98 kDa) are in green and actin (∼42 kDa) in red. Sample size: n = 7–10/treatment/sex/region. Abbreviations: GLUR1, AMPA receptor subunit R1; GLUR2, AMPA receptor subunit R2; HIPP, hippocampus; NA, nucleus accumbens; STR, striatum. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Antibodies were rabbit anti-NMDA-NR1 (Ab109182, AbCam, Cambridge, MA) at 1:4,000 with Odyssey IRDye 800 secondary antibody at 1:3,000 dilution; rabbit anti-NMDA-NR2A (Ab124913, AbCam, Cambridge, MA) at 1:9,000 with Odyssey IRDye 800 secondary antibody at 1:20,000 dilution; rabbit anti-NMDA-NR2B (Ab81271, AbCam, Cambridge, MA) at 1:5,000 with Odyssey IRDye 800 secondary antibody at 1:20,000 dilution; rabbit anti-ionotropic glutamate receptor 1 (GluR1) (Ab109450, AbCam, Cambridge, MA) at 1:9,000 with Odyssey IRDye 800 secondary antibody at 1:15,000 dilution; rabbit anti-ionotropic glutamate receptor 2 (GluR2) (Ab133477, AbCam, Cambridge, MA) at 1:7,000 with Odyssey IRDye 800 secondary antibody at 1:10,000 dilution; rabbit anti-DRD1 (Ab40653, AbCam, Cambridge, MA) at 1:1,000 with Odyssey IRDye 800 secondary antibody at 1:3,000 dilution; rabbit anti-DRD2 (Ab85367, AbCam, Cambridge, MA) at 1:500 with Odyssey IRDye 800 secondary antibody at 1:3,000 dilution; rabbit anti-DAT (Ab184451, AbCam, Cambridge, MA) at 1:2,000 with Odyssey IRDye 800 secondary antibody at 1:20,000 dilution; and mouse anti-β Actin (P/N: 926-42212, LI-COR Biosciences, Lincoln, NE) at 1:2,000 with Odyssey IRDye 680 secondary antibody at 1:15,000 dilution.

Techniques: Western Blot, Expressing

Journal: Molecular Vision

Article Title: Diabetes changes the levels of ionotropic glutamate receptors in the rat retina

doi:

Figure Lengend Snippet: Primary antibodies used in this study

Article Snippet: Rabbit anti-GluR2 , 1:500 , BD Biosciences (Heidelberg, Germany).

Techniques:

Journal: Molecular Vision

Article Title: Diabetes changes the levels of ionotropic glutamate receptors in the rat retina

doi:

Figure Lengend Snippet: Primers for qPCR analysis.

Article Snippet: Rabbit anti-GluR2 , 1:500 , BD Biosciences (Heidelberg, Germany).

Techniques:

Journal: Molecular Vision

Article Title: Diabetes changes the levels of ionotropic glutamate receptors in the rat retina

doi:

Figure Lengend Snippet: Effect of diabetes on the protein levels of AMPA receptor subunits. Total retinal extracts were obtained from rat retinas with diabetes for one, four, and 12 weeks (black bars) and from age-matched control rats (white bars). Extracts were assayed for ( A ) GluR1, ( B ) GluR2, ( C ) GluR2/3, and ( D ) GluR4 subunits immunoreactivity by western blot analysis. Representative western blots are presented above the bars for each time point tested. The densitometry of each band was analyzed. The results are expressed as percentage of age-matched controls and are presented as the mean±SEM, for the indicated number of animals. In each western blot analysis, a reprobing for detection of α-tubulin was performed to confirm that similar amounts of protein were applied to the gel. The asterisk indicates a p<0.05, the double asterisk indicates a p<0.01, and the triple asterisk indicates a p<0.001, significantly different from control, using the two-tailed Student’s t -test.

Article Snippet: Rabbit anti-GluR2 , 1:500 , BD Biosciences (Heidelberg, Germany).

Techniques: Western Blot, Two Tailed Test

Journal: Molecular Vision

Article Title: Diabetes changes the levels of ionotropic glutamate receptors in the rat retina

doi:

Figure Lengend Snippet: Effect of diabetes on the transcript levels of AMPA receptor subunits. Total RNA was isolated from rat retinas with one, four and 12 weeks of diabetes (black bars) and from rat retinas of age-matched controls (white bars). The transcript levels of ( A ) GluR1, ( B ) GluR2, ( C ) GluR3, ( D ) GluR4 subunits were analyzed by qPCR. The results represent the normalized expression levels for each subunit, as explained in Methods, and are presented as the mean±SEM, for the indicated number of animals. The asterisk indicates a p<0.05, significantly different from control, using the two-tailed Student’s t -test.

Article Snippet: Rabbit anti-GluR2 , 1:500 , BD Biosciences (Heidelberg, Germany).

Techniques: Isolation, Expressing, Two Tailed Test

Journal: Molecular Vision

Article Title: Diabetes changes the levels of ionotropic glutamate receptors in the rat retina

doi:

Figure Lengend Snippet: Ratios of the flip:flop and short:long splice variants for AMPA receptor subunits

Article Snippet: Rabbit anti-GluR2 , 1:500 , BD Biosciences (Heidelberg, Germany).

Techniques: