Journal: Schizophrenia (Heidelberg, Germany)

Article Title: FMR1 genetically interacts with DISC1 to regulate glutamatergic synaptogenesis.

doi: 10.1038/s41537-024-00532-7

Figure Lengend Snippet: Fig. 3 DGluRIIA expression in the control and DISC1OE NMJs. a NMJs of muscles 6-7 in the second abdominal segment were stained with anti-HRP (green) and anti-DGluRIIA (magenta). Bar, 20 μm. b Quantification of DGluRIIA protein expression level normalized to HRP immunoreactivity in dfmr1null/+ (dfmr1Δ50M or dfmr1Δ113M) NMJ boutons with (+) or without (−) DISC1 over- expression (DISC1OE). Kruskal–Wallis test (p = 0.0832) followed by Dunn’s multiple comparisons (w (CS10); DISC1OE (−) vs. w (CS10); DISC1OE (+), p = 0.0372, z = 2.677. w (CS10); DISC1OE (−) vs. dfmr1Δ50M; DISC1OE (−), p = 0.8535, z = 1.370. w (CS10); DISC1OE (−) vs. dfmr1Δ113M; DISC1OE (−), p = 0.0661, z = 2.478. dfmr1Δ50M; DISC1OE (−) vs. dfmr1Δ50M; DISC1OE (+), p > 0.9999, z = 0.8286. dfmr1Δ113M; DISC1OE (−) vs. dfmr1Δ113M; DISC1OE (+), p > 0.9999, z = 0.7481). Significance levels in the figures are represented as p < 0.05 (*) and n.s. not significant. n = 5–9. Individual values are plotted in the graphs. Data are presented as the mean ± SEM.

Article Snippet: In the present study, the following antibodies were used: sheep anti-DISC1 antibody diluted 1:50 (AF6699, R&D systems, Minneapolis, MN, USA), mouse anti-synaptotagmin diluted 1:2 (3H2 2D7, Developmental Studies Hybridoma Bank (DSHB), University of Iowa, IA, USA), mouse anti-Drosophila GluRIIA diluted 1:50 (8B4D2, DSHB), mouse anti-Futsch diluted 1:2 (22C10, DSHB), mouse anti-Brp64,66 diluted 1:20 (NC82, DSHB), antihorseradish peroxidase protein (HRP) conjugated with fluorescein-isothiocyanate diluted 1:50 (Jackson ImmunoResearch, West Grove, PA, USA), and Alexa-conjugated secondary antibodies diluted 1:1000 (Molecular probes, Eugene, OR, USA).

Techniques: Expressing, Control, Muscles, Staining, Over Expression

Journal: Schizophrenia (Heidelberg, Germany)

Article Title: FMR1 genetically interacts with DISC1 to regulate glutamatergic synaptogenesis.

doi: 10.1038/s41537-024-00532-7

Figure Lengend Snippet: Fig. 4 Brp expression in the control and DISC1OE NMJs. a NMJs of muscles 6-7 in the second abdominal segment were stained with anti-HRP (green) and anti-Brp (magenta). Bar, 20 μm. b Quantification of Brp protein expression level normalized to HRP immunoreactivity in dfmr1null/+ (dfmr1Δ50M or dfmr1Δ113M) NMJ boutons with (+) or without (−) DISC1 overexpression (DISC1OE). One-way ANOVA (F (5, 36) = 10.94, p < 0.0001) followed by Holm–Sidak’s multiple comparisons (w (CS10); DISC1OE (−) vs. w (CS10); DISC1OE (+), p = 0.0039, t = 3.591, df = 36. w (CS10); DISC1OE (−) vs. dfmr1Δ50M; DISC1OE (−), p = 0.9395, t = 0.0764, df = 36. w (CS10); DISC1OE (−) vs. dfmr1Δ113M; DISC1OE (−), p = 0.0025, t = 3.828, df = 36. dfmr1Δ50M; DISC1OE (−) vs. dfmr1Δ50M; DISC1OE (+), p = 0.2730, t = 1.481, df = 36. dfmr1Δ113M; DISC1OE (−) vs. dfmr1Δ113M; DISC1OE (+), p = 0.0509, t = 2.497, df = 36). Significance levels in the figures are represented as p < 0.01 (**) and n.s. not significant. n = 4–10. Individual values are plotted in the graphs. Data are presented as the mean ± SEM.

Article Snippet: In the present study, the following antibodies were used: sheep anti-DISC1 antibody diluted 1:50 (AF6699, R&D systems, Minneapolis, MN, USA), mouse anti-synaptotagmin diluted 1:2 (3H2 2D7, Developmental Studies Hybridoma Bank (DSHB), University of Iowa, IA, USA), mouse anti-Drosophila GluRIIA diluted 1:50 (8B4D2, DSHB), mouse anti-Futsch diluted 1:2 (22C10, DSHB), mouse anti-Brp64,66 diluted 1:20 (NC82, DSHB), antihorseradish peroxidase protein (HRP) conjugated with fluorescein-isothiocyanate diluted 1:50 (Jackson ImmunoResearch, West Grove, PA, USA), and Alexa-conjugated secondary antibodies diluted 1:1000 (Molecular probes, Eugene, OR, USA).

Techniques: Expressing, Control, Muscles, Staining, Over Expression

Journal: Schizophrenia (Heidelberg, Germany)

Article Title: FMR1 genetically interacts with DISC1 to regulate glutamatergic synaptogenesis.

doi: 10.1038/s41537-024-00532-7

Figure Lengend Snippet: Fig. 5 Futsch expression in the control and DISC1OE NMJs. a NMJs of muscles 6-7 in the second abdominal segment were stained with anti-HRP (green) and anti-Futsch (magenta). Bar, 20 μm. b Quantification of Futsch protein expression level normalized to HRP immunoreactivity in dfmr1null/+ (dfmr1Δ50M or dfmr1Δ113M) NMJ boutons with (+) or without (−) DISC1 overexpression (DISC1OE). One-way ANOVA (F (5, 63) = 4.620, p = 0.0012) followed by Holm–Sidak’s multiple comparisons (w (CS10); DISC1OE (−) vs. w (CS10); DISC1OE (+), p = 0.4444, t = 0.8837, df = 63. w (CS10); DISC1OE (−) vs. dfmr1Δ50M; DISC1OE (−), p = 0.4444, t = 1.324, df = 63. w (CS10); DISC1OE (−) vs. dfmr1Δ113M; DISC1OE (−), p = 0.0002, t = 4.380, df = 63. dfmr1Δ50M; DISC1OE (−) vs. dfmr1Δ50M; DISC1OE (+), p = 0.4444, t = 1.362, df = 63. dfmr1Δ113M; DISC1OE (−) vs. dfmr1Δ113M; DISC1OE (+), p = 0.0182, t = 2.940, df = 63). Significance levels in the figures are represented as p < 0.05 (*), p < 0.001 (***), and n.s. not significant. n = 5–16. Individual values are plotted in the graphs. Data are presented as the mean ± SEM.

Article Snippet: In the present study, the following antibodies were used: sheep anti-DISC1 antibody diluted 1:50 (AF6699, R&D systems, Minneapolis, MN, USA), mouse anti-synaptotagmin diluted 1:2 (3H2 2D7, Developmental Studies Hybridoma Bank (DSHB), University of Iowa, IA, USA), mouse anti-Drosophila GluRIIA diluted 1:50 (8B4D2, DSHB), mouse anti-Futsch diluted 1:2 (22C10, DSHB), mouse anti-Brp64,66 diluted 1:20 (NC82, DSHB), antihorseradish peroxidase protein (HRP) conjugated with fluorescein-isothiocyanate diluted 1:50 (Jackson ImmunoResearch, West Grove, PA, USA), and Alexa-conjugated secondary antibodies diluted 1:1000 (Molecular probes, Eugene, OR, USA).

Techniques: Expressing, Control, Muscles, Staining, Over Expression

Journal: Frontiers in Synaptic Neuroscience

Article Title: Stimulation of Synaptic Vesicle Exocytosis by the Mental Disease Gene DISC1 is Mediated by N-Type Voltage-Gated Calcium Channels

doi: 10.3389/fnsyn.2016.00015

Figure Lengend Snippet: DISC1 loss-of-function reduces evoked Ca 2+ transients at nerve terminals . SyGC3 imaging in rat hippocampal neurons (DIV14–16) in response two different trains of APs. (A) Average Ca 2+ transients in neurons expressing scr (1052 boutons, 15 fields, 5 experiments (exps)), DISC1-E (1836 boutons, 9 fields, 3 exps) and -A (754 boutons, 11 fields, 3 exps) shRNAs, in response to 300 APs, 10 Hz. The DISC1-E and DISC-A groups are significantly different than the scr group ( p = 0.0057). (B) Average Ca 2+ transients in neurons expressing scr (861 boutons, 9 fields, 5 exps), DISC1-E (2118 boutons, 7 fields, 3 exps) and -A (1549 boutons, 9 fields, 3 exps) shRNAs, in response to 200 APs, 20 Hz. (C) Cumulative probability of SyGC3 peak intensity from individual boutons corresponding to (A) . (D) Average Ca 2+ transients in DISC1 wt/wt (1090 boutons, 11 fields, 3 exps) and DISC1 Δ2–3/Δ2–3 (727 boutons, 9 fields, 3 exps) neurons, in response to 300 APs, 10 Hz. The DISC1 wt/wt and DISC1 Δ2–3/Δ2–3 groups are statistically different ( p = 0.0182). (E) Average Ca 2+ transients in DISC1 wt/wt (1365 boutons, 9 fields, 3 exps) and DISC1 Δ2–3/Δ2–3 (1233 boutons, 8 fields, 3 exps) neurons, in response to 200 APs, 20 Hz. (F) Cumulative probability of SyGC3 peak intensity from individual boutons corresponding to (D) . (G) Average Ca 2+ transients in neurons expressing scr (290 boutons, 6 fields, 2 exps), and DISC1-E (390 boutons, 6 fields, 2 exps) shRNAs, in response to 20 APs, 20 Hz. The scr and DISC1-E groups are statistically different ( p = 0.0036). (H) Average vGpH traces in scr (180 boutons, 5 fields, 2 exps) and DISC1-E (488 boutons, 6 fields, 2 exps) shRNA-expressing neurons during two consecutive trains of APs (300 AP, 10 Hz) in the presence of 2 or 4 mM extracellular Ca 2+ .

Article Snippet: The human DISC1 gene L variant (NCBI Refseq NM018662.2) was PCR amplified from pCMV6-XL5 DISC1-tGFP (Origene) and cloned into pIRES2-DsRed-Express (Clontech) using NheI and SmaI sites.

Techniques: Imaging, Expressing, shRNA

Journal: Frontiers in Synaptic Neuroscience

Article Title: Stimulation of Synaptic Vesicle Exocytosis by the Mental Disease Gene DISC1 is Mediated by N-Type Voltage-Gated Calcium Channels

doi: 10.3389/fnsyn.2016.00015

Figure Lengend Snippet: DISC1 silencing by RNAi slows down SV exocytosis. (A) Immunoblot analysis of DISC1 in mouse hippocampal neurons (DIV 8) transduced with scramble, DISC1-A and DISC1-E shRNAs. (B) Average vGpH traces derived from neurons expressing scr (1204 boutons, 18 fields), -E (960 boutons, 18 fields) and -A (924 boutons, 16 fields) shRNAs from six independent experiments. (C,D) Analysis of SV exocytosis after Baf treatment. (C) Exocytic profiles of neurons expressing the indicated shRNAs. (D) Boxplot of exocytic rates. (E,F) Analysis of SV endocytosis. (E) Endocytic profiles. (F) Boxplot of endocytic rates. (G) Exocytic profile of cells expressing the indicated shRNAs in response to a stimulation (1200 APs, 10 Hz) that depletes the total releasable pool.

Article Snippet: The human DISC1 gene L variant (NCBI Refseq NM018662.2) was PCR amplified from pCMV6-XL5 DISC1-tGFP (Origene) and cloned into pIRES2-DsRed-Express (Clontech) using NheI and SmaI sites.

Techniques: Western Blot, Transduction, Derivative Assay, Expressing

Journal: Frontiers in Synaptic Neuroscience

Article Title: Stimulation of Synaptic Vesicle Exocytosis by the Mental Disease Gene DISC1 is Mediated by N-Type Voltage-Gated Calcium Channels

doi: 10.3389/fnsyn.2016.00015

Figure Lengend Snippet: Attenuated SV release in hippocampal neurons from DISC1 Δ2–3/Δ2–3 mice. (A) Immunoblot analysis of DISC1 in hippocampal lysates prepared from P10 DISC1 Δ2–3/Δ2–3 , DISC1 wt/Δ 2–3 and DISC1 wt/wt mice, confirming the ablation of full-length DISC1 (~100 kD). (B) Average vGpH traces in DISC1 wt/wt (575 boutons, 7 fields) and DISC1 Δ2–3/Δ2–3 (682 boutons, 9 fields) neurons in response to two consecutive trains of APs and obtained from two independent experiments. (C,D) Analysis of SV exocytosis from vGpH responses after Baf. Average kinetics (C) and rates (D) of vGpH exocytic responses. (E,F) Analysis of SV endocytosis. Average kinetics (E) and rates (F) of vGpH endocytic responses.

Article Snippet: The human DISC1 gene L variant (NCBI Refseq NM018662.2) was PCR amplified from pCMV6-XL5 DISC1-tGFP (Origene) and cloned into pIRES2-DsRed-Express (Clontech) using NheI and SmaI sites.

Techniques: Western Blot

Journal: Frontiers in Synaptic Neuroscience

Article Title: Stimulation of Synaptic Vesicle Exocytosis by the Mental Disease Gene DISC1 is Mediated by N-Type Voltage-Gated Calcium Channels

doi: 10.3389/fnsyn.2016.00015

Figure Lengend Snippet: DISC1 regulates Cav2.2-dependent SV exocytosis. (A) Average vGpH traces in scr (101 boutons, 6 fields, 2 exps) and DISC1-E (87 boutons, 5 fields, 2 exps) shRNA-expressing neurons during consecutive trains of APs (300 AP, 10 Hz) in the absence or presence of the Cav2.2 blocker ω-Conotoxin GVIA (125 nM). (B) Boxplot of SV exocytic rates before and after ω-Conotoxin GVIA application. Exocytic rates were measured by linear fitting of the first six time points of the vGpH response. (C) Average vGpH traces in scr (1294 boutons, 9 fields, 3 exps) and DISC1-E (1282 boutons, 9 fields, 3 exps) shRNA-expressing neurons during consecutive trains of APs (300 AP, 10 Hz) in the absence or presence of the Cav2.1 blocker ω-Agatoxin TK (125 nM). (D) Boxplot of SV exocytic rates before and after ω-Agatoxin TK application. (E) Table showing the percentage of inhibition of exocytosis rate by DISC1 knockdown before and after Cav2.2- or Cav2.1 blockade.

Article Snippet: The human DISC1 gene L variant (NCBI Refseq NM018662.2) was PCR amplified from pCMV6-XL5 DISC1-tGFP (Origene) and cloned into pIRES2-DsRed-Express (Clontech) using NheI and SmaI sites.

Techniques: shRNA, Expressing, Inhibition, Knockdown

Journal: Frontiers in Synaptic Neuroscience

Article Title: Stimulation of Synaptic Vesicle Exocytosis by the Mental Disease Gene DISC1 is Mediated by N-Type Voltage-Gated Calcium Channels

doi: 10.3389/fnsyn.2016.00015

Figure Lengend Snippet: DISC1 enhances Cav2.2 and Cav2.1 currents. (A) Western blot showing expression of ectopic (human) DISC1 in HEK293 cells. (B,C) Cav2.2 Current-voltage (I-V) curves for hDISC1-expressing and control cells. (B) Stimulation protocol and individual current responses shown at three different voltages (−30, 0 and 30 mV) (C) . Average I-V plots for hDISC1 (peak = 54.2 ± 4.5 pA/pF, 13 cells) and control (peak = 39.2 ± 4.9 pA/pF, 12 cells), p = 0.033. (D–F) Cav2.2 activation curves in response to the tail protocol. (D) Illustration of the tail protocol and individual tail currents measured at −50 mV after three different voltage steps (−20, 0 and 40 mV). (E) Average current density based on tail currents for DISC1 (164.3 ± 7.3 pA/pF, 12 cells) and control (110.5 ± 6.3 pA/pF, 12 cells), * p < 0.001. (F) Normalized activation curve from tail currents showing no significant difference between DISC1 (V 50 : −6.98 ± 3.43 mV, 12 cells) and control (V 50 : 0.86 ± 3.62 mV, 12 cells), p = 0.13. (G,H) Cav2.1 Current-voltage (I-V) curves for hDISC1-expressing and control cells. (G) Stimulation protocol and individual current responses shown at three different voltages (−30, 0 and 30 mV). (H) Average I-V plots for hDISC1 (peak = 79.9 ± 8.3 pA/pF, 15 cells) and control (peak = 56.3 ± 7.4 pA/pF, 13 cells), p = 0.046. (I–K) Cav2.1 activation curves in response to the tail protocol. (I) Illustration of the tail protocol and individual tail currents measured at −50 mV after three different voltage steps (−20, 0 and 40 mV). (J) Average current density based on tail currents for DISC1 (156.3 ± 5.7 pA/pF, 25 cells) and control (123.2 ± 7.4 pA/pF, 17 cells), * p = 0.003. (K) Normalized activation curve from tail currents showing no significant difference between DISC1 (V 50 : −5.39 ± 0.49 mV, 24 cells) and control (V 50 : −4.41 ± 0.93 mV, 17 cells), p = 0.31.

Article Snippet: The human DISC1 gene L variant (NCBI Refseq NM018662.2) was PCR amplified from pCMV6-XL5 DISC1-tGFP (Origene) and cloned into pIRES2-DsRed-Express (Clontech) using NheI and SmaI sites.

Techniques: Western Blot, Expressing, Control, Activation Assay

Journal: Nature Communications

Article Title: DISC1 Protects Against Zika Virus Infection and Long-Term Neurological Damage Through AMPK-mTOR-Mediated Autophagy

doi: 10.1038/s41467-025-64809-w

Figure Lengend Snippet: a Venn diagram showing the overlap of differentially expressed genes (DEGs) identified through transcriptomic and proteomic analyses of ZIKV-infected HTR8 cells compared to non-infected controls. The overlapping area indicates shared DEGs between the two datasets. b − d HTR8 cells were transfected with pcDNA or HA-DISC1 (1 μg) plasmids for 24 hours ( h ) and then infected with ZIKV at a multiplicity of infection (MOI) of 1. Viral mRNA levels, protein expression and titers were assessed on day 1 (D1) and day 2 (D2) post-infection using qRT-PCR ( b ), Western blot ( c ), and plaque assay ( d ). e − g U251 cells were transfected with pcDNA or HA-DISC1 (1 μg) plasmids for 24 h and infected with ZIKV at a MOI of 1. Viral mRNA levels, protein expression and titers were measured on D1 and D2 post-infection using qRT-PCR ( e ), Western blot ( f ), and plaque assay ( g ). h , i HTR8 cells ( h ) or U251 cells ( i ) were transfected with siRNA targeting DISC1 for 24 h prior to ZIKV infection at a MOI of 1. Viral protein was analyzed on D1 and D2 post-infection using Western blot. j − m HTR8 cells ( j , k ) or U251 cells ( l , m ) were transfected with siRNA targeting DISC1 for 24 h prior to ZIKV infection at a MOI of 1. Viral mRNA levels and viral loads were analyzed on D1 and D2 post-infection using qRT-PCR ( j , l ) and plaque assay ( k , m ). Data shown in ( b , d – e , g , j – m ) are from one representative experiment out of three independent replicates with similar results, each including 3 biological replicates per group ( n = 3). Images in ( c − i ) are from one representative experiment out of three independent replicates with similar results ( n = 3). All the data were confirmed to follow a normal distribution using the Shapiro-Wilk test, and statistical analysis was performed using Student’s unpaired two-tailed t test. Data are presented as means ± SD. Source data are provided as a Source Data file.

Article Snippet: The Disc1 knockdown mice (C57BL/6 background) were generated by Cyagen Biosciences (Guangzhou, China) using CRISPR/Cas9-mediated deletion of exon 2 of the Disc1 gene.

Techniques: Infection, Transfection, Expressing, Quantitative RT-PCR, Western Blot, Plaque Assay, Two Tailed Test

Journal: Nature Communications

Article Title: DISC1 Protects Against Zika Virus Infection and Long-Term Neurological Damage Through AMPK-mTOR-Mediated Autophagy

doi: 10.1038/s41467-025-64809-w

Figure Lengend Snippet: a Schematic representation of the experiment set up. 6-8-week-old WT or Disc1 KD mice were treated with 2 mg MAR1-5A3 on the day prior to infection and then intraperitoneally (i.p.) inoculated with PBS or 1 × 10 6 PFU of ZIKV. Mouse tissues were collected on D2 and D6 post-infection. b − h Viral titers in plasma ( b ) and uterus ( h ) were determined by plaque assay, and mRNA levels in blood cell ( c ), spleen ( d ), brain ( e ), testis ( f ) and uterus ( g ) were measured by qRT-PCR on D2 and D6 post-infection. i , j Primary cells isolated from WT or Disc1 KD mice were induced to differentiate into macrophages using mouse macrophage colony-stimulating factor. Cells were pre-incubated with 20 μg/mL anti-IFNAR1 antibody MAR1-5A3 for 6 h, followed by infection with ZIKV at a MOI of 1. Viral mRNA levels and titers were measured on D1 and D3 post-infection using qRT-PCR ( i ) and plaque assay ( j ). Data for plasma ( b ), blood cells ( c ), spleen ( d ), and brain ( e ) are one representative experiment out of two independent replicates with similar results, each including 6 mice per group ( n = 6), consisting of 3 males and 3 females. Data for the testis ( f ) are one representative experiment out of two independent replicates with similar results, each including 6 male mice per group ( n = 6). Data for the uterus ( g , h ) are one representative experiment out of two independent replicates with similar results, each including 6 female mice per group ( n = 6). Data shown in ( i , j ) are one representative experiment out of two independent replicates with similar results, each including 4 biological replicates per group ( n = 4). Data shown in ( b , e , j ) did not follow a normal distribution according to the Shapiro-Wilk test, and statistical analysis was performed using two-tailed Mann-Whitney test. Data shown in ( c – d , f – i ) were confirmed to follow a normal distribution using the Shapiro-Wilk test, and statistical analysis was performed using Student’s unpaired two-tailed t test. Data are presented as means ± SD. Source data are provided as a Source Data file. Figure 2a was created in BioRender. Hl, Z. (2025) https://BioRender.com/bh2c5ad .

Article Snippet: The Disc1 knockdown mice (C57BL/6 background) were generated by Cyagen Biosciences (Guangzhou, China) using CRISPR/Cas9-mediated deletion of exon 2 of the Disc1 gene.

Techniques: Infection, Clinical Proteomics, Plaque Assay, Quantitative RT-PCR, Isolation, Incubation, Two Tailed Test, MANN-WHITNEY

Journal: Nature Communications

Article Title: DISC1 Protects Against Zika Virus Infection and Long-Term Neurological Damage Through AMPK-mTOR-Mediated Autophagy

doi: 10.1038/s41467-025-64809-w

Figure Lengend Snippet: a Schematic representation of the experimental setup. One day prior to infection, 2 mg of the anti-IFNAR1 antibody MAR1-5A3 was administered to 8-10-week-old WT dams and Disc1 KD dams on E5.5. On E6.5, dams were i.p. inoculated with either PBS or 1 × 10 7 PFU of ZIKV. Placenta and fetal head were harvested on E13.5. b Representative images of E13.5 uteri (upper panel) and fetuses (lower panel). Partial demise and growth restriction was shown in ZIKV-infected WT and Disc1 KD pregnant dams. Red arrows indicate the placental residues, and green arrows show the growth restriction of fetuses. c Resorption rates were analyzed on E13.5. d− g The weight ( d ), CRL ( e ), OFD ( f ) and size ( g , CRL × OFD) of fetuses were measured on E13.5. h , i ZIKV mRNA levels in placentas ( h ) and fetal heads ( i ) of WT and Disc1 KD mice was measured by qRT-PCR. j ZIKV mRNA levels in the spleen of in WT or Disc1 KD dams were measured by qRT-PCR. Data shown in ( d − i ) are one representative experiment out of two independent replicates with similar results, each including 4 pregnant dams per group ( n = 4). The offspring analyzed were NF-WT ( n = 36), NF-KD ( n = 37), ZIKV-WT ( n = 28), and ZIKV-KD ( n = 20). Data shown in j are one representative experiment out of two independent replicates with similar results, each including 4 pregnant dams per group ( n = 4). Data shown in ( d − i ) did not follow a normal distribution according to the Shapiro-Wilk test, and statistical analysis was performed using two-tailed Kruskal-Wallis test followed by Dunn’s multiple comparison test ( d − g ) or two-tailed Mann-Whitney test ( h , i ). Data shown in e and j were confirmed to follow a normal distribution using the Shapiro-Wilk test, and statistical analysis was performed using two-tailed one-way ANOVA followed by Tukey’s multiple comparison test ( e ) or Student’s unpaired two-tailed t test ( j ). Data are presented as means ± SD. Source data are provided as a Source Data file. Figure 3a was created in BioRender. Hl, Z. (2025) https://BioRender.com/dvzm7vs .

Article Snippet: The Disc1 knockdown mice (C57BL/6 background) were generated by Cyagen Biosciences (Guangzhou, China) using CRISPR/Cas9-mediated deletion of exon 2 of the Disc1 gene.

Techniques: Infection, Quantitative RT-PCR, Two Tailed Test, Comparison, MANN-WHITNEY

Journal: Nature Communications

Article Title: DISC1 Protects Against Zika Virus Infection and Long-Term Neurological Damage Through AMPK-mTOR-Mediated Autophagy

doi: 10.1038/s41467-025-64809-w

Figure Lengend Snippet: a Schematic representation of the experiment set up. On the second day after birth, WT and Disc1 KD neonatal mice were intracranially injected with 200 PFU of ZIKV or an equal volume of PBS. Brains were collected on D3, D6 and D9 post-infection, as well as at 6 weeks of age. b ZIKV mRNA levels in WT and Disc1 KD brains were assessed by qRT-PCR on D3, D6 and D9 post-infection, as well as at 6 weeks of age. c , d Body weight ( c ) and brain weight ( d ) of WT and Disc1 KD mice were measured on D3, D6, D9 post-infection, as well as at 6 weeks of age. e , f Representative images of WT and Disc1 KD mice brains on D3, D6, D9 post-infection ( e ), as well as at 6 weeks of age ( f ). The yellow scale bar indicates consistent size within the same age group. g The cerebral size of WT and Disc1 KD mice was measured on D3, D6, D9 post-infection, as well as at 6 weeks of age. Data shown in ( b − d , g ) are collected from two independent replicates, including 8 mice per group ( n = 8), consisting of 4 males and 4 females. Images in ( e , f ) are one representative experiment out of two independent replicates with similar results. Data shown in ( b ) did not follow a normal distribution according to the Shapiro-Wilk test, and statistical analysis was performed using two-tailed Mann-Whitney test. Data shown in ( c – d , g ) were confirmed to follow a normal distribution using the Shapiro-Wilk test, and statistical analysis was performed using two-tailed one-way ANOVA followed by Tukey’s multiple comparison test. Data are presented as means ± SD. Source data are provided as a Source Data file. Figure 4a was created in BioRender. Hl, Z. (2025) https://BioRender.com/s3be69y .

Article Snippet: The Disc1 knockdown mice (C57BL/6 background) were generated by Cyagen Biosciences (Guangzhou, China) using CRISPR/Cas9-mediated deletion of exon 2 of the Disc1 gene.

Techniques: Injection, Infection, Quantitative RT-PCR, Two Tailed Test, MANN-WHITNEY, Comparison

Journal: Nature Communications

Article Title: DISC1 Protects Against Zika Virus Infection and Long-Term Neurological Damage Through AMPK-mTOR-Mediated Autophagy

doi: 10.1038/s41467-025-64809-w

Figure Lengend Snippet: a On the second day after birth, WT and Disc1 KD neonatal mice were intracranially injected with 200 PFU of ZIKV or an equal volume of PBS, and behavioral tests were conducted when the mice reached 6 weeks of age. The spontaneous alternation rate of WT and Disc1 KD mice in the Y-maze test. b The passing time of WT and Disc1 KD mice in the balance beam test. c , d Proportion of entries into the open arms ( c ) and the percentage of time spent in the open arms ( d ) by WT and Disc1 KD mice in the elevated plus maze test. e , f The time of WT and Disc1 KD mice spent on socializing with Stranger 1 ( e ) and Stranger 2 ( f ) during the 3-chamber test. Data shown in ( a − d ) are collected from one independent experiment, each including 8 mice per group ( n = 8), consisting of 4 males and 4 females. Data shown in ( e , f ) are collected from one independent experiment, each including 10 mice per group ( n = 10), consisting of 5 males and 5 females. Data shown in ( a - b , d – f ) were confirmed to follow a normal distribution using the Shapiro-Wilk test, and statistical analysis was performed using two-tailed one-way ANOVA followed by Tukey’s multiple comparison test. Data shown in c did not follow a normal distribution according to the Shapiro-Wilk test, statistical analysis was performed using two-tailed Kruskal-Wallis test followed by Dunn’s multiple comparison test. Data are presented as means ± SD. Source data are provided as a Source Data file.

Article Snippet: The Disc1 knockdown mice (C57BL/6 background) were generated by Cyagen Biosciences (Guangzhou, China) using CRISPR/Cas9-mediated deletion of exon 2 of the Disc1 gene.

Techniques: Injection, Two Tailed Test, Comparison

Journal: Nature Communications

Article Title: DISC1 Protects Against Zika Virus Infection and Long-Term Neurological Damage Through AMPK-mTOR-Mediated Autophagy

doi: 10.1038/s41467-025-64809-w

Figure Lengend Snippet: a KEGG pathway enrichment of DEGs in ZIKV-infected U251 cells transfected with HA-DISC1 (1 μg) plasmid compared with the control group transfected with pcDNA. b KEGG pathway enrichment of differentially expressed proteins identified by IP/MS analysis in U251 cells overexpressing HA-DISC1 (5 μg) plasmid compared with the control group transfected with pcDNA. c Chord diagram illustrates the proteins identified in the IP/MS analysis. d U251 cells were transfected with pcDNA or HA-DISC1 (1 μg) plasmids for 24 h followed by ZIKV infection. The protein levels of AMPKα, pAMPKα, mTOR, pmTOR, P62 and LC3A/B were assessed by Western blot at 0, 12, 24, and 36 h post-infection. e U251 cells were transfected with pcDNA or HA-DISC1 (1 μg) plasmids for 24 h, followed by ZIKV infection for 6 h and treated with DMSO, rapamycin (25 μM), and MHY1485 (10 μM) for 24 h. ZIKV E expression was assessed by Western blot. f U251 cells were transfected with pcDNA or HA-DISC1 (1 μg) plasmids for 24 h, followed by ZIKV infection for 6 h and treated with DMSO, autophagosome inhibitor 3-MA (1 mg/ml), and lysosome inhibitor CQ (100 μM) for 24 h. ZIKV E expression was assessed by Western blot. KEGG pathway enrichment analyses in ( a , b ) were performed using a one-tailed Fisher’s exact test. Images in ( d − f ) are one representative experiment out of three independent replicates with similar results ( n = 3). Source data are provided as a Source Data file.

Article Snippet: The Disc1 knockdown mice (C57BL/6 background) were generated by Cyagen Biosciences (Guangzhou, China) using CRISPR/Cas9-mediated deletion of exon 2 of the Disc1 gene.

Techniques: Infection, Transfection, Plasmid Preparation, Control, Protein-Protein interactions, Western Blot, Expressing, One-tailed Test

Journal: Nature Communications

Article Title: DISC1 Protects Against Zika Virus Infection and Long-Term Neurological Damage Through AMPK-mTOR-Mediated Autophagy

doi: 10.1038/s41467-025-64809-w

Figure Lengend Snippet: a Conserved LIR motifs (amino acids 210 FSFI 213) in DISC1 were highlighted in yellow, which were mutated into AAAA in DISC1 mutant. b 293T cells were co-transfected with HA-DISC1 (5 μg) or HA-DISC1 mutant (5 μg) and GFP-LC3 (5 μg) plasmids for 48 h. Cellular lysates were subjected to immunoprecipitation with anti-Flag or anti-HA magnetic beads and Western blot assays using the indicated antibodies. c − e U251 cells were transfected with HA-DISC1 (1 μg) or HA-DISC1 mutant (1 μg) plasmids for 24 h, followed by ZIKV infection for 24 and 48 h. Viral mRNA levels, protein expression and titers were measured on D1 and D2 post-infection by qRT-PCR ( c ), Western blot ( d ), and plaque assays ( e ). Images in ( b , d ) are from one representative experiment out of three independent replicates with similar results ( n = 3). Data shown in ( c , e ) are from one representative experiment out of three independent replicates with similar results, each including 3 biological replicates per group ( n = 3). Data shown in ( c , e ) were confirmed to follow a normal distribution using the Shapiro-Wilk test, and statistical analysis was performed using two-tailed one-way ANOVA followed by Tukey’s multiple comparison test. Data are presented as means ± SD. Source data are provided as a Source Data file.

Article Snippet: The Disc1 knockdown mice (C57BL/6 background) were generated by Cyagen Biosciences (Guangzhou, China) using CRISPR/Cas9-mediated deletion of exon 2 of the Disc1 gene.

Techniques: Mutagenesis, Transfection, Immunoprecipitation, Magnetic Beads, Western Blot, Infection, Expressing, Quantitative RT-PCR, Two Tailed Test, Comparison

Journal: Nature Communications

Article Title: DISC1 Protects Against Zika Virus Infection and Long-Term Neurological Damage Through AMPK-mTOR-Mediated Autophagy

doi: 10.1038/s41467-025-64809-w

Figure Lengend Snippet: a Primary cortical neurons were isolated from neonatal mice (P0–P1) and induced to differentiate in vitro. On day 7 of differentiation, neurons exhibiting neurite outgrowth were observed under bright-field microscopy. Confocal imaging showed neuronal nuclei stained with DAPI (blue), and endogenous Neun labeled with an anti-Neun antibody (red). Scale bar = 50 μm. b , c WT and Disc1 KD primary cortical neurons were pre-incubated with 20 μg/mL MAR1-5A3 for 6 h, followed by infection with ZIKV at a MOI of 1. Viral mRNA levels and titers were measured on D1 and D2 post-infection using qRT-PCR ( b ) and plaque assay ( c ). d WT and Disc1 KD primary cortical neurons were pre-incubated with 20 μg/mL MAR1-5A3 for 6 h, followed by infection with ZIKV at a MOI of 1. The protein levels of ZIKV E protein, DISC1, AMPKα, pAMPKα, mTOR, pmTOR, P62 and LC3A/B were assessed by Western blot on D1 and D2 post-infection. e, f The ratios of pAMPKα to AMPKα ( e ) and pmTOR to mTOR ( f ) were determined by comparing the corresponding protein band intensities at each time point in Fig. 8d. g − i Quantification of P62 ( g ), LC3A/BⅠ ( h ) and LC3A/BⅡ ( i ) protein levels relative to GAPDH at each time point in Fig. 8d. j , k Placentas ( j ) and fetal heads ( k ) from non-infected and ZIKV-infected WT and Disc1 KD mice were collected on E13.5. The protein levels of DISC1, AMPKα, pAMPKα, mTOR, pmTOR, P62 and LC3A/B were assessed by Western blot. l − o The ratios of pAMPKα to AMPKα ( l, m ) and pmTOR to mTOR ( n , o ) were determined by comparing the corresponding protein band intensities at each time point in Fig. 8j ( l, n ) and Fig. 8k ( m , o ). p − s Quantification of P62 ( p , q ) and LC3A/BⅡ ( r , s ) protein levels relative to GAPDH at each time point in Fig. 8j ( p , r ) and Fig. 8k ( q , s ). Data shown in ( b , c ) are from one representative experiment out of three independent replicates with similar results, each including 3 biological replicates per group ( n = 3). Images in ( a , d , j – k ) are from one representative experiment out of three independent replicates with similar results ( n = 3). Data shown in ( e − i , l − s) are collected from three independent experiments ( n = 3). Data shown in ( b − c , e − i , l − s ) were confirmed to follow a normal distribution using the Shapiro-Wilk test, and statistical analysis was performed using Student’s unpaired two-tailed t test ( b , c ) or two-tailed one-way ANOVA followed by Tukey’s multiple comparison test ( e − i , l − s ). Data are presented as means ± SD. Source data are provided as a Source Data file.

Article Snippet: The Disc1 knockdown mice (C57BL/6 background) were generated by Cyagen Biosciences (Guangzhou, China) using CRISPR/Cas9-mediated deletion of exon 2 of the Disc1 gene.

Techniques: Isolation, In Vitro, Microscopy, Imaging, Staining, Labeling, Incubation, Infection, Quantitative RT-PCR, Plaque Assay, Western Blot, Two Tailed Test, Comparison

Journal: Nature Communications

Article Title: DISC1 Protects Against Zika Virus Infection and Long-Term Neurological Damage Through AMPK-mTOR-Mediated Autophagy

doi: 10.1038/s41467-025-64809-w

Figure Lengend Snippet: a 6-8-weeks-old WT or Disc1 KD mice were treated with 2 mg MAR1-5A3 on the day prior to infection and then intraperitoneally (i.p.) inoculated with PBS or 1 × 10 6 PFU of ZIKV. Mouse brains were collected on D6 post-infection. Representative H&E-stained sections of the hippocampal DG and CA3 regions are shown. Black arrows indicate shrunken, hyperchromatic neurons with indistinct boundaries Scale bar = 100 μm. b Representative immunofluorescence-stained sections of the hippocampal DG region. Cell nuclei were stained using DAPI (blue). Endogenous Neun was labeled with anti-Neun antibody (red). Endogenous LC3A/B was labeled with anti-LC3A/B antibody (yellow). Scale bar = 100 μm. Representative images in ( a , b ) are from one independent experiment, including 6 mice per group ( n = 6), consisting of 3 males and 3 females.

Article Snippet: The Disc1 knockdown mice (C57BL/6 background) were generated by Cyagen Biosciences (Guangzhou, China) using CRISPR/Cas9-mediated deletion of exon 2 of the Disc1 gene.

Techniques: Infection, Staining, Immunofluorescence, Labeling

Journal: Life Science Alliance

Article Title: Kalirin-RAC controls nucleokinetic migration in ADRN-type neuroblastoma

doi: 10.26508/lsa.201900332

Figure Lengend Snippet: (A) Immunolabelling of kalirin in IMR-32 after KALRN RNAi. Scale bar 20 μm. (B) Western blot analysis of kalirin-8 and kalirin-STYV levels in IMR-32 and SK-N-BE(2)c after KALRN RNAi and kalirin-8 levels in IMR32 after KALRN RNAi or additional negative control siRNAs: control siRNA #2, HNRNPK siRNA, and DISC1 siRNA. (C) RAC1 activity in IMR-32 and SK-N-BE(2)c cell treated with kalirin-GEF1 inhibitor#1 (10 μM), kalirin-GEF1 inhibitor#2 (5 μM), RHOA inhibitor (3 μM), RAC1 inhibitor (10 μM), or after SOX11 and KALRN RNAi. (D) Kalrn expression in t-SNE-resolved E12.5 and E13.5 sympathetic precursors: sympathoblasts, Schwann cell precursors, bridge population and chromaffin cells . (E) Sequencing electrophoregrams showing 3′-UTR of kalirin-9 isoform (top) and the exon scheme based on the results of sequencing (bottom). Location of stop codon is marked with “-.” (F) KALRN , TRIO , and TIAM expression in SK-N-BE(2)c (left) after RA (10 μM) treatment. x-axis indicate timepoints in hours. Western Blot analysis of kalirin, TRIO, and βIII-tubulin in SK-N-BE2c after 72 h of RA-treatment (top right) and of TRIO and kalirin-8 in IMR-32 and SK-N-BE(2)c after 72 h of RA treatment (bottom right). (G) Cell viability of NB cell lines treated with vehicle, kalirin-GEF1 inhibitor#2, or kalirin-GEF1 inhibitor#1. Values are reported as mean percent ± SD of vehicle-treated control.

Article Snippet: KALRN (sc-18592), LIS1 (sc-35814), DCX (sc-35214), HNRNPK (sc-38282), and DISC1 (sc-60539) were purchased from Santa Cruz Biotechnologies.

Techniques: Western Blot, Negative Control, Control, Activity Assay, Expressing, Sequencing

Journal: The Journal of Biological Chemistry

Article Title: DISC1-dependent Regulation of Mitochondrial Dynamics Controls the Morphogenesis of Complex Neuronal Dendrites *

doi: 10.1074/jbc.M115.699447

Figure Lengend Snippet: DISC1 interacts with mitochondrial trafficking complex proteins to regulate transport in dendrites in addition to axons. A , GFP trap co-immunoprecipitation experiments from COS7 cells show robust interaction of DISC1 with GFP Miro1, GFP Miro2, GFP TRAK1, and GFP TRAK2. B , proximity ligation assay in SH-SY5Y cells with DISC1 antibody or a DISC1 and Miro1 antibody shows significantly increased signal in the dual antibody condition over background, indicating that DISC1 and Miro1 interact within the cell ( n = 3 individual preparations). Scale bar = 20 μm. IB , immunoblot; IN , input. C and D , co-immunoprecipitation experiments with rat brain homogenate showing DISC1 to be part of a native complex with Miro ( C ) and TRAK1 ( n = 3) or TRAK2 ( n = 3) ( D ). E , example of live labeling of the axon initial segment ( AIS ) to distinguish axons and dendrites within the same neuron. F , kymographs showing movement of mitochondria through the axons and dendrites over time. Moving mitochondria are indicated by diagonal lines and stationary mitochondria by straight lines. G , dendritic compartments ( gray bars ) and axonal compartments ( black bars ) were assayed for mitochondrial movement, and the percentage of moving mitochondria was quantified with and without expression of DISC1 (dendrites: n = 15 control neurons and n = 16 DISC1 neurons, *, p = 0.02; axons: n = 16 control neurons and n = 13 DISC1 neurons, *, p = 0.02). Scale bar = 5 μm. NS , not significant.

Article Snippet: Anti-human DISC1 (14F2) was described previously (WB and immunofluorescence 1:100 mouse) ( ); anti-Rhot1 against Miro was from Atlas Antibodies (HPA010687 for proximity ligation assays and AMAb90854 for immunoprecipitation), and anti-TRAK1 was also from Atlas Antibodies (HPA005853).

Techniques: Immunoprecipitation, Proximity Ligation Assay, Western Blot, Labeling, Expressing, Control

Journal: The Journal of Biological Chemistry

Article Title: DISC1-dependent Regulation of Mitochondrial Dynamics Controls the Morphogenesis of Complex Neuronal Dendrites *

doi: 10.1074/jbc.M115.699447

Figure Lengend Snippet: DISC1 is recruited to mitochondria by components of the mitochondrial trafficking complex. A , immunocytochemistry in COS7 cells showing localization of exogenous DISC1 with and without myc Miro1 overexpression. Mitochondria are labeled with MtDsRed2. Scale bar = 20 μm. B , percentage of DISC1 on mitochondria shown in A ( n = 13–15 cells from three individual experiments, ***, p = 2.29 × 10 −5 ). Overexpression of myc Miro1 recruits DISC1 to mitochondria. C , mitochondrial fractionation from COS7 cells shows an increase in DISC1 in this compartment with GFP TRAK1 and myc Miro overexpression ( n = 3). IN , input.

Article Snippet: Anti-human DISC1 (14F2) was described previously (WB and immunofluorescence 1:100 mouse) ( ); anti-Rhot1 against Miro was from Atlas Antibodies (HPA010687 for proximity ligation assays and AMAb90854 for immunoprecipitation), and anti-TRAK1 was also from Atlas Antibodies (HPA005853).

Techniques: Immunocytochemistry, Over Expression, Labeling, Fractionation

Journal: The Journal of Biological Chemistry

Article Title: DISC1-dependent Regulation of Mitochondrial Dynamics Controls the Morphogenesis of Complex Neuronal Dendrites *

doi: 10.1074/jbc.M115.699447

Figure Lengend Snippet: The DISC1 N terminus mediates the interaction with Miro and TRAKs. A , schematic of the DISC1 protein showing domains present in deletion constructs used. Coiled-coil domains are dark blue , and nuclear import or export signals are dark red. B , schematic of TRAK2 showing coiled-coil domains ( dark blue ). C and D , mapping the region of DISC1 that interacts with myc Miro1 ( C ) and GFP TRAK2 ( D ). Co-IP experiments from COS7 cells show that the N-terminal 301 amino acids interact with Miro and TRAK2, whereas amino acids 313–854 are not pulled down. Arrowhead , highlights full-length ( FL ) HA DISC1 band; *, indicates nonspecific band. E and F , mapping the region of TRAK2 which interacts with DISC1. DISC1 interacts with full-length TRAK2 and TRAK2-(1–700) ( E ) but does not interact with the Miro binding domain ( F ). G , kymographs showing effect of overexpressing the DISC1-Miro binding domain on mitochondrial transport. Scale bar = 10 μm. H , percentage of moving mitochondria quantified in axons and dendrites. Overexpression of the DISC1-Miro binding domain prevents mitochondrial transport ( n = 23 ctrl and n = 21 DISC1-(1–301)-expressing neurons from three preparations, **, p = 0.002). I , quantification of percentage of moving mitochondria in axons and dendrites expressing MtDsRed2 (ctrl) or DISC1-(313–854), the region that does not interact with Miro1 ( n = 12 ctrl and 11 DISC1-(313–854)-expressing neurons from three preparations, p = 0.6). IB , immunoblot; IN , input; NS , not significant.

Article Snippet: Anti-human DISC1 (14F2) was described previously (WB and immunofluorescence 1:100 mouse) ( ); anti-Rhot1 against Miro was from Atlas Antibodies (HPA010687 for proximity ligation assays and AMAb90854 for immunoprecipitation), and anti-TRAK1 was also from Atlas Antibodies (HPA005853).

Techniques: Construct, Co-Immunoprecipitation Assay, Binding Assay, Over Expression, Expressing, Western Blot

Journal: The Journal of Biological Chemistry

Article Title: DISC1-dependent Regulation of Mitochondrial Dynamics Controls the Morphogenesis of Complex Neuronal Dendrites *

doi: 10.1074/jbc.M115.699447

Figure Lengend Snippet: The DISC1-Boymaw fusion protein inhibits mitochondrial trafficking. A , immunocytochemistry in hippocampal neurons showing localization of the Boymaw protein. Scale bar = 20 μm, 5 μm on zoomed image ( right ). B , line scan of zoomed process showing that the DISC1-Boymaw fusion protein localizes to mitochondria in axons and dendrites. ( A.U. = arbitrary units.) C , kymographs showing mitochondrial transport in axons expressing MtDsRed2 and co-expressing HA Boymaw. Scale bar = 10 μm. D , percentage of moving mitochondria in neurons was quantified with and without expression of HA Boymaw. The presence of the HA Boymaw fusion protein inhibits mitochondrial trafficking ( n = 32 ctrl neurons and 26 HA Boymaw-expressing neurons, ***, p = 0.001). E and F , Boymaw expression has no impact on synaptophysin trafficking ( E ), as quantified in F ( n = 17 control and n = 19 HA Boymaw-expressing neurons from three preparations, p = 0.9). NS , not significant. Scale bar = 10 μm.

Article Snippet: Anti-human DISC1 (14F2) was described previously (WB and immunofluorescence 1:100 mouse) ( ); anti-Rhot1 against Miro was from Atlas Antibodies (HPA010687 for proximity ligation assays and AMAb90854 for immunoprecipitation), and anti-TRAK1 was also from Atlas Antibodies (HPA005853).

Techniques: Immunocytochemistry, Expressing, Control

Journal: The Journal of Biological Chemistry

Article Title: DISC1-dependent Regulation of Mitochondrial Dynamics Controls the Morphogenesis of Complex Neuronal Dendrites *

doi: 10.1074/jbc.M115.699447

Figure Lengend Snippet: DISC1-mediated mitochondrial trafficking is necessary for normal dendritic development. A , representative images showing control 10 DIV neurons and those expressing the DISC1-Miro binding domain (residues 1–301). GFP was used to visualize neuronal morphology. Scale bar = 10 μm. B , total dendritic length/cell (***, p = 0.001). C , Sholl analysis reveals a significant decrease in intersections at 80 and 100 μm from the soma (*, p < 0.05). D , average number of branch points/cell is decreased with DISC1-Miro binding domain expression (*, p = 0.04). E , Sholl analysis of branch points (*, p < 0.05 at 90 μm from the soma) ( n = 16 neurons from four preparations). F , representative images showing control 10 DIV neurons expressing GFP in the presence or absence of HA Boymaw. Scale bar = 10 μm. G , total dendritic length is decreased when the DISC1-Boymaw fusion protein is expressed (**, p = 0.004). H , Sholl analysis showing the number of intersections at 10-μm intervals. A significant decrease in intersections in Boymaw-expressing neurons is seen proximal to the soma at a distance of 50–80 μm away from the soma (*, p < 0.05 in each case). I , analysis of branch points reveals expression of the DISC1-Boymaw fusion protein to decrease in the number of branch points/cell (*, p = 0.03). J , Sholl analysis reveals a significant decrease 50 μm from the soma (*, p < 0.05, n = 15–16 neurons from four preparations).

Article Snippet: Anti-human DISC1 (14F2) was described previously (WB and immunofluorescence 1:100 mouse) ( ); anti-Rhot1 against Miro was from Atlas Antibodies (HPA010687 for proximity ligation assays and AMAb90854 for immunoprecipitation), and anti-TRAK1 was also from Atlas Antibodies (HPA005853).

Techniques: Control, Expressing, Binding Assay

Journal: The Journal of Biological Chemistry

Article Title: DISC1-dependent Regulation of Mitochondrial Dynamics Controls the Morphogenesis of Complex Neuronal Dendrites *

doi: 10.1074/jbc.M115.699447

Figure Lengend Snippet: DISC1 interacts with mitofusins. A and B , DISC1 1–301 decreases the length of mitochondria ( A ) as quantified in B (ctrl = 2. 1 μm ± 0.065, DISC1-(1–301) = 1.8 ± 0.063 μm, n = 11 axons, **, p = 0.004). C and D , the DISC1 Boymaw fusion protein decreases the length of mitochondria (ctrl = 1.81 ± 0.0858 μm, HA Boymaw = 1.54 ± 0.0644 μm, n = 13 axons, *, p = 0.02). Scale bar = 10 μm. E , co-IP experiments from COS7 cells show that DISC1 interacts with Myc Mitofusin1 and -2. F , mitochondrial fractionation from COS7 cells shows that DISC1 is recruited to mitochondria upon overexpression of Myc Mitofusin1 and -2. G , co-IP experiments with rat brain homogenate showing DISC1 to be part of a native complex with Mitofusin1 but not with translocase component TOM20 ( n = 3). IB , immunoblot; IN , input.

Article Snippet: Anti-human DISC1 (14F2) was described previously (WB and immunofluorescence 1:100 mouse) ( ); anti-Rhot1 against Miro was from Atlas Antibodies (HPA010687 for proximity ligation assays and AMAb90854 for immunoprecipitation), and anti-TRAK1 was also from Atlas Antibodies (HPA005853).

Techniques: Co-Immunoprecipitation Assay, Fractionation, Over Expression, Western Blot

Journal: The Journal of Biological Chemistry

Article Title: DISC1-dependent Regulation of Mitochondrial Dynamics Controls the Morphogenesis of Complex Neuronal Dendrites *

doi: 10.1074/jbc.M115.699447

Figure Lengend Snippet: The DISC1-Boymaw fusion protein inhibits mitochondrial fusion. A , HA Boymaw inhibits mitochondrial fusion in neurons. Neurons were transfected with MtDsRed2 and mitochondrially targeted photoactivatable GFP (ctrl) or coexpressing HA Boymaw. Scale bar = 20 μm. B , the change in area of GFP signal after photoactivation is reduced in HA Boymaw-expressing neurons ( n = 17 control and n = 15 Boymaw-expressing neurons, p < 0.05 at 12–90 s post-photoactivation, **, p < 0.01 at 96–570 s post-photoactivation). C , schematic showing mitochondrial fusion after polyethylene glycol-mediated plasma membrane fusion. D and E , HA Boymaw decreases mitochondrial fusion in COS7 cells ( D ) as quantified by colocalization analysis in E ( n = 15 post-fusion cells from three individual preparations, ***, p = 0.0009). Scale bar = 20 μm.

Article Snippet: Anti-human DISC1 (14F2) was described previously (WB and immunofluorescence 1:100 mouse) ( ); anti-Rhot1 against Miro was from Atlas Antibodies (HPA010687 for proximity ligation assays and AMAb90854 for immunoprecipitation), and anti-TRAK1 was also from Atlas Antibodies (HPA005853).

Techniques: Transfection, Expressing, Control, Clinical Proteomics, Membrane

Journal: The Journal of Biological Chemistry

Article Title: DISC1-dependent Regulation of Mitochondrial Dynamics Controls the Morphogenesis of Complex Neuronal Dendrites *

doi: 10.1074/jbc.M115.699447

Figure Lengend Snippet: The DISC1-Boymaw fusion protein decreases the area of ER-mitochondria contacts. A , three-dimensional renderings of mitochondrial network (Su9 GFP ) and ER (ER dsred ) in COS7 cells upon co expression of HA DISC1 or HA Boymaw. Colocalization shows regions of ER-mitochondria interface. Scale bar = 20 μm, 5 μm on zoomed images ( right hand panel ). B , colocalization of HA DISC1 or HA Boymaw with ER-mitochondria contacts. The images of the contact sites were generated from colocalization of images shown in A. C , Manders colocalization coefficient of images in A shows a decrease in area of ER-mitochondria contacts upon HA Boymaw expression ( n = 15 cells from three experiments, ctrl versus Boymaw p < 0.05). NS , not significant. D , Manders coefficient for colocalization of HA DISC1 or HA Boymaw with ER-mitochondria contact sites. HA Boymaw shows higher colocalization than HA DISC1 (*, p = 0.03). E , structured illumination microscopy showing ER and mitochondrial network in SH-SY5Y cells. F , structured illumination microscopy shows partial colocalization between endogenous DISC1 and ER-mitochondria contacts in SH-SY5Y cells. Scale bar = 10 μm (zoom 1 μm). G , line scan showing signal intensities of DISC1 and ER-mitochondria contacts. A.U. = arbitrary units.

Article Snippet: Anti-human DISC1 (14F2) was described previously (WB and immunofluorescence 1:100 mouse) ( ); anti-Rhot1 against Miro was from Atlas Antibodies (HPA010687 for proximity ligation assays and AMAb90854 for immunoprecipitation), and anti-TRAK1 was also from Atlas Antibodies (HPA005853).

Techniques: Expressing, Generated, Microscopy

Journal: Neurobiology of disease

Article Title: Hippocampal Pyk2 regulates specific social skills: Implications for schizophrenia.

doi: 10.1016/j.nbd.2024.106487

Figure Lengend Snippet: Fig. 5. Levels of hippocampal Pyk2 and mitochondrial markers in patients with schizophrenia. (a) Immunoblot densitometric quantification of Pyk2 levels relative to α-actin levels as the loading control in dorsal hippocampal post-mortem samples from human schizophrenic subjects (SCZ) and matched controls (CNT). (Mann-Whitney U test; A, B = 306, 1585, U = 201; p = 0.0276). (b-d) Fixed hippocampal tissues from 3 CNT and 4 SCZ were subjected to DAPI staining (b) and Pyk2 immunofluorescence (c). (d) Densitometric quantification of Pyk2 immunofluorescence in the samples in c in arbitrary units (unpaired t-test; t = 11.25, df = 70, p < 0.0001). A total of 32 CNT and 40 SCZ pyramidal neurons were quantified (10–14 per subject). Immunoblot densitometric quantification of DISC1 (e and f, Mann-Whitney U test; A, B = 135, 243, U = 44; p = 0.0222), Mfn2 (e and g; Mann-Whitney U test; A, B = 154, 252, U = 49; p = 0.0241), and VDAC1 (e and h) levels relative to α-actin levels as the loading control in dorsal hippocampal post-mortem samples from human schizophrenic subjects (SCZ) and matched controls (CNT). Immunoblot densitometric quantification of Grp75 (i and j; Mann-Whitney U test; A, B = 133, 273, U = 28; p = 0.0008) and VDAC1 (i and k; unpaired t-test; t = 2.243, df = 26, p = 0.036) levels relative to α-actin levels as the loading control in dorsal hippocampal post- mortem samples from human schizophrenic subjects (SCZ) and matched controls (CNT). Data represent the mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001 vs CNT. In a, e-k, n = 14/group.

Article Snippet: Immunoblots were probed with the following antibodies (all diluted 1:1000): Pyk2 (Sigma, #074 M4755), phospho-Y402-Pyk2 (Invitrogen, #44- 618G), Src (Abcam, #ab4705), phospho-Y418-Src (Abcam, #ab47411), CoxV (Invitrogen, #A21347), OXPHOS cocktail (Abcam, #ab110413), DISC1 (Proteintech, #15500–1-AP), Mfn2 (Abcam, #ab56889), VDAC1 (Abcam, #ab15895), Grp75 (Abcam, #ab2799), ACSL4 (Proteintech, #22401–1-AP), and phospho-tyrosine (4G10) mouse mAb (Cell Signaling, #96215).

Techniques: Western Blot, Control, MANN-WHITNEY, Staining, Immunofluorescence