Journal: Nature Communications

Article Title: Enhanced TARP-γ8-PSD-95 coupling in excitatory neurons contributes to the rapid antidepressant-like action of ketamine in male mice

doi: 10.1038/s41467-023-42780-8

Figure Lengend Snippet: a Social interaction ratio of SIT ( n = 10, 10, 11, and 10 in CON-Vehicle, CON-Ketamine, CSDS-Vehicle and CSDS-Ketamine, respectively). b Time in the interaction zone of SIT ( n = 10, 10, 12, and 10 in CON-Vehicle, CON-Ketamine, CSDS-Vehicle and CSDS-Ketamine, respectively). c Preference for sucrose in the SPT ( n = 9, 9, 10, and 10 in CON-Vehicle, CON-Ketamine, CSDS-Vehicle and CSDS-Ketamine, respectively). d , e Immobility time in the FST ( d ) and total distance in the OFT ( e ) ( n = 10, 10, 12, and 12 in CON-Vehicle, CON-Ketamine, CSDS-Vehicle and CSDS-Ketamine, respectively). f Representative traces of AMPARs-mediated mEPSC recordings in ventral hippocampal CA1 neurons. g Quantification of cumulative probability and amplitude and frequency of mEPSCs ( n = 9 cells from 4 mice in CON-Vehicle, n = 8 cells from 3 mice in CON-Ketamine, n = 9 cells from 3 mice in CSDS-Vehicle, n = 8 cells from 4 mice in CSDS-Ketamine, respectively). h Representative western blot image of Co-IP assay. i Quantification of association between PSD-95 and TARP-γ8 in cultured primary hippocampal neurons ( n = 7, 8, and 7 in 0, 1 h and 24 h, respectively). j , k Quantification of total expression of PSD-95 ( j ) and TARP-γ8 ( k ) in cultured primary hippocampal neurons ( n = 7, 8, and 8 in 0, 1 h, and 24 h, respectively). l Representative western blot image of Co-IP assay. m Quantification of association between PSD-95 and TARP-γ8 in the ventral hippocampus ( n = 8, 7, 7, and 7 in CON-Vehicle, CON-Ketamine, CSDS-Vehicle and CSDS-Ketamine, respectively). n , o Quantification of total expression of PSD-95 ( n ) and TARP-γ8 ( o ) in the ventral hippocampus ( n = 8 per group). Comparisons were performed by two-way ANOVA analysis followed by Bonferroni’s multiple comparisons test in ( a– g , m – o ) and one-way ANOVA analysis followed by Dunnett’s multiple comparisons test in i – k . Data are presented as mean ± SEM. All numbers ( n ) are biologically independent experiments. Source data are provided as a Source Data file.

Article Snippet: Primary antibodies used to recognize specific protein, including: TARP-γ8 (1:1000; sc-514421, Santa Cruz Biotechnology, Santa Cruz, CA), GluA1 (1:1000; ab31232, Abcam, Cambridge, UK), GluA2 (1:1000; ab52932, Abcam, Cambridge, UK), β-actin (1:3000; sc-47778, Santa Cruz Biotechnology, Santa Cruz, CA), PSD-95 (1:1000; ab18258, Abcam, Cambridge, UK), CaMKIIα (1:800; 50049, Cell Signaling, USA), Phospho-CaMKII (1:800; 12716, Cell Signaling, USA).

Techniques: Western Blot, Co-Immunoprecipitation Assay, Cell Culture, Expressing

Journal: Nature Communications

Article Title: Enhanced TARP-γ8-PSD-95 coupling in excitatory neurons contributes to the rapid antidepressant-like action of ketamine in male mice

doi: 10.1038/s41467-023-42780-8

Figure Lengend Snippet: a Schematic representation of LV-mediated TARP-γ8 or TARP-γ8-Δ4 overexpression and a schematic diagram of the experimental process. b Targeted locations and confocal images of GFP (green) expression in the ventral hippocampus. Scale bar = 50 μm (left), 500 μm (right). Experiments were repeated independently 3 times with similar results. c Social interaction ratio of SIT ( n = 9, 11, 11, 11, 11, and 13 in CON-LV-GFP, CSDS-LV-GFP, CON-LV- Cacng8 , CSDS-LV- Cacng8 , CON-LV- Cacng8 - Δ4 and CSDS-LV- Cacng8 - Δ4 , respectively). d Immobility time in the TST ( n = 9, 10, 11, 9, 10, and 12 in CON-LV-GFP, CSDS-LV-GFP, CON-LV- Cacng8 , CSDS-LV- Cacng8 , CON-LV- Cacng8 - Δ4 and CSDS-LV- Cacng8 - Δ4 , respectively). e Immobility time in the FST ( n = 10, 9, 11, 11, 9, and 11 in CON-LV-GFP, CSDS-LV-GFP, CON-LV- Cacng8 , CSDS-LV- Cacng8 , CON-LV- Cacng8 - Δ4 and CSDS-LV- Cacng8 - Δ4 , respectively). f Total distance in the OFT ( n = 9, 12, 11, 11, 11, and 12 in CON-LV-GFP, CSDS-LV-GFP, CON-LV- Cacng8 , CSDS-LV- Cacng8 , CON-LV- Cacng8 - Δ4 and CSDS-LV- Cacng8 - Δ4 , respectively). g The representative image of the western blot. h Quantification of protein expression of surface GluA1 ( n = 10, 12, 11, 11, 10, and 7 in CON-LV-GFP, CSDS-LV-GFP, CON-LV- Cacng8 , CSDS-LV- Cacng8 , CON-LV- Cacng8-Δ4 and CSDS-LV- Cacng8-Δ4 , respectively), surface GluA2 ( n = 10, 12, 11, 11, 10, and 10 in CON-LV-GFP, CSDS-LV-GFP, CON-LV- Cacng8 , CSDS-LV- Cacng8 , CON-LV- Cacng8-Δ4 and CSDS-LV- Cacng8-Δ4 , respectively) and TARP-γ8 ( n = 14, 12, 9, 9, 8, and 9 in CON-LV-GFP, CSDS-LV-GFP, CON-LV- Cacng8 , CSDS-LV- Cacng8 , CON-LV- Cacng8-Δ4 and CSDS-LV- Cacng8-Δ4 , respectively). i Representative traces of AMPARs-mediated mEPSC recordings from different groups. j – m Quantification of cumulative probability ( j , k ) and amplitude ( l ) and frequency ( m ) of mEPSCs ( n = 8 cells from 5 mice in CON-LV-GFP, n = 9 cells from 6 mice in CSDS-LV-GFP, n = 7 cells from 6 mice in CON-LV- Cacng8 , n = 8 cells from 5 mice in CSDS-LV- Cacng8 , n = 8 cells from 6 mice in CON-LV- Cacng8-Δ4 and n = 10 cells from 6 mice in CSDS-LV- Cacng8-Δ4 ). Comparisons were performed by two-way ANOVA analysis followed by Bonferroni’s multiple comparisons test. s/t stands for surface/total in h . Data are presented as mean ± SEM. All numbers ( n ) are biologically independent experiments. Source data are provided as a Source Data file.

Article Snippet: Primary antibodies used to recognize specific protein, including: TARP-γ8 (1:1000; sc-514421, Santa Cruz Biotechnology, Santa Cruz, CA), GluA1 (1:1000; ab31232, Abcam, Cambridge, UK), GluA2 (1:1000; ab52932, Abcam, Cambridge, UK), β-actin (1:3000; sc-47778, Santa Cruz Biotechnology, Santa Cruz, CA), PSD-95 (1:1000; ab18258, Abcam, Cambridge, UK), CaMKIIα (1:800; 50049, Cell Signaling, USA), Phospho-CaMKII (1:800; 12716, Cell Signaling, USA).

Techniques: Over Expression, Expressing, Western Blot

Journal: Nature Communications

Article Title: Enhanced TARP-γ8-PSD-95 coupling in excitatory neurons contributes to the rapid antidepressant-like action of ketamine in male mice

doi: 10.1038/s41467-023-42780-8

Figure Lengend Snippet: a AAV-expressing constructs encoding GFP and shRNAs targeting TARP-γ8 and region-specific expression of GFP in the ventral hippocampus of mice. b Targeted locations and confocal images of AAV-mediated GFP (green) expression in the ventral hippocampus. Scale bar = 100 μm (inside), 500 μm (outside). Experiments were repeated independently three times with similar results. c Quantification of TARP-γ8 protein expression ( n = 6 mice per group). d , e Immobility time in the TST ( d ) and FST ( e ) ( n = 11 mice in AAV- Scr -shRNA, n = 10 mice in AAV- Cacng8 -shRNA). f Total distance traveled in the OFT ( n = 11 mice in AAV- Scr -shRNA, n = 10 mice in AAV- Cacng8 -shRNA). g The representative image of western blot. h The quantification of total and surface protein expression of GluA1 and GluA2 ( n = 16 mice in AAV- Scr -shRNA, n = 10 mice in AAV- Cacng8 -shRNA). i Representative traces of AMPARs-mediated mEPSC recordings from different groups. j Whole-cell patch-clamp was used to record AMPARs-mediated mEPSC of CA1 neurons in the ventral hippocampus with GFP (bright) expression. Scale bar = 50 μm. k Quantification of cumulative probability and amplitude and frequency of mEPSCs ( n = 11 cells from four mice in AAV- Scr -shRNA, n = 13 cells from 6 mice in AAV- Cacng8 -shRNA). Comparisons were performed by unpaired, two-tailed t test. Data are presented as mean ± SEM. All numbers ( n ) are biologically independent experiments. Source data are provided as a Source Data file.

Article Snippet: Primary antibodies used to recognize specific protein, including: TARP-γ8 (1:1000; sc-514421, Santa Cruz Biotechnology, Santa Cruz, CA), GluA1 (1:1000; ab31232, Abcam, Cambridge, UK), GluA2 (1:1000; ab52932, Abcam, Cambridge, UK), β-actin (1:3000; sc-47778, Santa Cruz Biotechnology, Santa Cruz, CA), PSD-95 (1:1000; ab18258, Abcam, Cambridge, UK), CaMKIIα (1:800; 50049, Cell Signaling, USA), Phospho-CaMKII (1:800; 12716, Cell Signaling, USA).

Techniques: Expressing, Construct, shRNA, Western Blot, Patch Clamp, Two Tailed Test

Journal: Nature Communications

Article Title: Enhanced TARP-γ8-PSD-95 coupling in excitatory neurons contributes to the rapid antidepressant-like action of ketamine in male mice

doi: 10.1038/s41467-023-42780-8

Figure Lengend Snippet: a The representative image of western blot. b The quantification of protein expression of p-CaMKIIα and CaMKIIα ( n = 9 per group). c Preference for sucrose in the SPT ( n = 9, 8, 9, and 10 in Vehicle-ACSF, Vehicle-KN93, Ketamine-ACSF, and Ketamine-KN93, respectively). d , e Immobility time in the FST ( d ) and total distance in the OFT ( e ) ( n = 9, 9, 10, and 8 in Vehicle-ACSF, Vehicle-KN93, Ketamine-ACSF and Ketamine-KN93, respectively). f Representative western blot image of Co-IP assay. g Quantification of association between PSD-95 and TARP-γ8 ( n = 8, 7, 7, and 9 in Vehicle-ACSF, Vehicle-KN93, Ketamine-ACSF and Ketamine-KN93, respectively). h , i Total protein expression of PSD-95 ( h ) and TARP-γ8 ( i ) ( n = 8 samples per group). j Representative traces of AMPARs-mediated mEPSC recordings in ventral hippocampal CA1 neurons from different groups. k , l Quantification of cumulative probability and amplitude ( k ) and frequency ( l ) of mEPSCs ( n = 8 cells from 4 mice in Vehicle-ACSF, n = 7 cells from four mice in Vehicle-KN93, n = 7 cells from four mice in Ketamine-ACSF and n = 8 cells from 4 mice in Ketamine-KN93). m Representative western blot image of Co-IP assay. n Quantification of the association between PSD-95 and TARP-γ8 ( n = 7 in Tat-Scrambled, n = 8 in Tat-TTPV). o Preference for sucrose in the SPT ( n = 9, 9, 8 and 8 in Vehicle-Tat-Scrambled, Vehicle-Tat-TTPV, Ketamine-Tat-Scrambled and Ketamine-Tat-TTPV, respectively). p , q Immobility time in the FST ( p ) and total distance in the OFT ( q ) ( n = 10, 9, 8 and 8 in Vehicle-Tat-Scrambled, Vehicle-Tat-TTPV, Ketamine-Tat-Scrambled and Ketamine-Tat-TTPV, respectively). Comparisons were performed by unpaired, two-tailed t test in ( b , n ) and two-way ANOVA analysis followed by Bonferroni’s multiple comparisons test in ( c – l , o – q ). Data are presented as mean ± SEM. All numbers ( n ) are biologically independent experiments. Source data are provided as a Source Data file.

Article Snippet: Primary antibodies used to recognize specific protein, including: TARP-γ8 (1:1000; sc-514421, Santa Cruz Biotechnology, Santa Cruz, CA), GluA1 (1:1000; ab31232, Abcam, Cambridge, UK), GluA2 (1:1000; ab52932, Abcam, Cambridge, UK), β-actin (1:3000; sc-47778, Santa Cruz Biotechnology, Santa Cruz, CA), PSD-95 (1:1000; ab18258, Abcam, Cambridge, UK), CaMKIIα (1:800; 50049, Cell Signaling, USA), Phospho-CaMKII (1:800; 12716, Cell Signaling, USA).

Techniques: Western Blot, Expressing, Co-Immunoprecipitation Assay, Two Tailed Test

Journal: Nature Communications

Article Title: Enhanced TARP-γ8-PSD-95 coupling in excitatory neurons contributes to the rapid antidepressant-like action of ketamine in male mice

doi: 10.1038/s41467-023-42780-8

Figure Lengend Snippet: a Immunofluorescence results showing TARP-γ8 (green) was mainly expressed in CaMKIIα (purple)-expressing neurons rather than GAD67 (purple)-expressing neurons. Scale bar = 30 μm (two bars on the left), 50 μm (two bars on the right). DAPI (blue). Experiments were repeated independently 3 times with similar results. b AAV-expressing constructs with CaMKIIα promoter encoding GFP and shRNAs targeting TARP-γ8. c Timeline of experimental procedure. d Preference for sucrose in the SPT ( n = 8 samples per group). e Immobility time in the FST ( n = 8, 8, 9, and 10 in Vehicle- Scr -shRNA, Vehicle- Cacng8 -shRNA, Ketamine- Scr -shRNA and Ketamine- Cacng8 -shRNA, respectively). f Total distance in the OFT ( n = 8, 9, 10 and 10 in Vehicle- Scr -shRNA, Vehicle- Cacng8 -shRNA, Ketamine- Scr -shRNA and Ketamine- Cacng8 -shRNA, respectively). g Representative traces of AMPARs-mediated mEPSC recordings from excitatory neurons in the ventral hippocampus. h Quantification of cumulative probability and amplitude and frequency of mEPSCs ( n = 10 cells from 4 mice in Vehicle- Scr -shRNA, n = 9 cells from 4 mice in Vehicle- Cacng8 -shRNA, n = 7 cells from 4 mice in Ketamine- Scr -shRNA and n = 8 cells from 4 mice in Ketamine- Cacng8 -shRNA). Comparisons were performed by two-way ANOVA analysis followed by Bonferroni’s multiple comparisons test. Data are presented as mean ± SEM. All numbers ( n ) are biologically independent experiments. Source data are provided as a Source Data file.

Article Snippet: Primary antibodies used to recognize specific protein, including: TARP-γ8 (1:1000; sc-514421, Santa Cruz Biotechnology, Santa Cruz, CA), GluA1 (1:1000; ab31232, Abcam, Cambridge, UK), GluA2 (1:1000; ab52932, Abcam, Cambridge, UK), β-actin (1:3000; sc-47778, Santa Cruz Biotechnology, Santa Cruz, CA), PSD-95 (1:1000; ab18258, Abcam, Cambridge, UK), CaMKIIα (1:800; 50049, Cell Signaling, USA), Phospho-CaMKII (1:800; 12716, Cell Signaling, USA).

Techniques: Immunofluorescence, Expressing, Construct, shRNA

Journal: Nature Communications

Article Title: Enhanced TARP-γ8-PSD-95 coupling in excitatory neurons contributes to the rapid antidepressant-like action of ketamine in male mice

doi: 10.1038/s41467-023-42780-8

Figure Lengend Snippet: Ketamine exhibits antidepressant effects by increasing the postsynaptic recruitment of TARP-γ8 and the enhanced excitatory synaptic transmission mediated by TARP-γ8-selective AMPAR in a CaMKII phosphorylation-dependent manner in the ventral hippocampus. Knockdown of TARP-γ8 in excitatory neurons of the ventral hippocampus or blockage of TARP-γ8-selective AMPAR abolishes the rapid antidepressant effects of ketamine.

Article Snippet: Primary antibodies used to recognize specific protein, including: TARP-γ8 (1:1000; sc-514421, Santa Cruz Biotechnology, Santa Cruz, CA), GluA1 (1:1000; ab31232, Abcam, Cambridge, UK), GluA2 (1:1000; ab52932, Abcam, Cambridge, UK), β-actin (1:3000; sc-47778, Santa Cruz Biotechnology, Santa Cruz, CA), PSD-95 (1:1000; ab18258, Abcam, Cambridge, UK), CaMKIIα (1:800; 50049, Cell Signaling, USA), Phospho-CaMKII (1:800; 12716, Cell Signaling, USA).

Techniques: Transmission Assay, Phospho-proteomics, Knockdown

Journal: Cell Proliferation

Article Title: Inhibition of L‐type voltage‐gated calcium channel‐mediated Ca 2+ influx suppresses the collective migration and invasion of ameloblastoma

doi: 10.1111/cpr.13305

Figure Lengend Snippet: Verapamil suppressed in vivo expansion and invasion of AM‐1 spheroids. (A) Timeline for AM‐1 cell line‐based orthotopic xenograft mouse model. (B) Extraction sites of first right molars in micro‐CT data were set as region of interest and bone volume to total volume (BV/TV) was measured ( n = 7). (C) Micro‐CT images, haematoxylin and eosin staining (HE), and immunofluorescence staining for human leukocyte antigen (HLA) of maxillae of xenograft mouse models. (D and E) Immunohistochemistry staining for cytokeratin 14 (KRT14), CACNA1C, and P‐cadherin. Nuclei were counterstained with TO‐PRO‐3 (TP3). Scale bars, C, HE, 500 μm; HLA, 100 μm; D, 100 μm; E, 50 μm. ** p < 0.0001

Article Snippet: The AM cells were transfected with either negative control siRNA (Scrambled; sc‐398433; Santa Cruz) or CACNA1C siRNA (sc‐42688; Santa Cruz) following the manufacturer's protocol for lipofectamine RNAiMAX (Life Technologies).

Techniques: In Vivo, Extraction, Micro-CT, Staining, Immunofluorescence, Immunohistochemistry