Journal: Translational Psychiatry

Article Title: CHD8 adulthood microglial knockdown in C57BL6 mice induces behavioral, morphological, and transcriptional changes in a sex-dependent manner

doi: 10.1038/s41398-025-03468-3

Figure Lengend Snippet: a Timeline of tamoxifen administration and animal sacrifice of two animal models for immunohistochemistry analysis. b, c Immunohistochemistry of the infralimbic cortex with antibodies targeting microglia (IbaI positive) and CHD8 in male b and female c mice. Nuclei are stained in blue (Hoechst). d, e Immunohistochemistry of hippocampus with antibodies targeting neurons (NeuN), and CHD8 in male d and female e mice. Nuclei are stained in blue (Hoechst).

Article Snippet: To induce conditional knockdown of Chd8 in excitatory neurons, the CamKiia-Cre-Ert C57BL6 mouse line (kindly provided by the laboratory of Prof. Alon Chen, Weizmann Institute of Science, Israel) was crossed with Chd8 lx/flx C57BL6 mice (The Jackson Laboratory, Strain #:031555) to generate the Chd8 lx/flx Camk2aCreErt +/− cKO mice.

Techniques: Immunohistochemistry, Staining

Journal: Translational Psychiatry

Article Title: CHD8 adulthood microglial knockdown in C57BL6 mice induces behavioral, morphological, and transcriptional changes in a sex-dependent manner

doi: 10.1038/s41398-025-03468-3

Figure Lengend Snippet: Chd8 -imKO males : a Time spent in the center of the open field. (* p = 0.0205, t = 2.554, df = 17, two-way student’s t-test) b Time spent in the light area of the dark-light exploration test. (* p = 0.0473, t = 2.149, df = 16, two-way student’s t-test) c Time spent in the open arm of the elevated plus maze (* p = 0.0242, t = 2.527, df = 14, two-way student’s t-test). d Time self-grooming within a 10 min time period. e Number of marbles buried in 30 min time period. f Time spent immobile in tail suspension test (* p = 0.0195, t = 2.580, df = 17, two-way student’s t-test) g Time spent immobile in forced swim test (** p = 0.0037, t = 3.437, df = 15, two-way student’s t-test). h Motor coordination and balance were assessed using the rotarod. Time spent before falling off rotarod. i Percent freezing in each of three tones in cue fear conditioning test (Two way ANOVA, Genotype effect: p = 0.0055, F(1, 17) = 10.09, Tukey’s post hoc test, * p = 0.0172, ** p = 0.0016, * p = 0.0173). j Time spent in interaction zone with stranger mice and empty chamber in three-chamber social interaction test (Two way ANOVA, Stranger effect: p < 0.0001, F(1, 62) = 22.68, Tukey’s post hoc test, *** p < 0.0001, ns p = 0.1589). n = wt 9–11 imKO 7–8. * p < 0.05, ** p < 0.01, *** p < 0.001. Chd8 -imKO females : k Time spent in the center of the open field (* p = 0.0239, t = 2.479, df = 17, two-way student’s t-test). l Time spent in the light area of the dark-light exploration test. m Time spent in the open arm of the elevated plus maze. n Time self-grooming within a 10 min time period. o Number of marbles buried in 30 min time period. p Time spent immobile in tail suspension test q Time spent immobile in forced swim test. r Motor coordination and balance were assessed using the rotarod. Time spent before falling off rotarod. s Percent freezing in each of three tones in cue fear conditioning test t Time spent in interaction zone with stranger mice and empty chamber in three-chamber social interaction test (Two way ANOVA, Stranger effect: p < 0.0001, F(1, 34) = 24.59, Tukey’s post hoc test * p = 0.0169, ** p = 0.0027). Further tests determined no differences between experimental groups. (l- dark-light, m- elevated plus maze, p- tail suspension test, q- forced swimming test). n = wt 9–10 imKO 9–10. * p < 0.05, ** p < 0.01.

Article Snippet: To induce conditional knockdown of Chd8 in excitatory neurons, the CamKiia-Cre-Ert C57BL6 mouse line (kindly provided by the laboratory of Prof. Alon Chen, Weizmann Institute of Science, Israel) was crossed with Chd8 lx/flx C57BL6 mice (The Jackson Laboratory, Strain #:031555) to generate the Chd8 lx/flx Camk2aCreErt +/− cKO mice.

Techniques: Suspension

Journal: Translational Psychiatry

Article Title: CHD8 adulthood microglial knockdown in C57BL6 mice induces behavioral, morphological, and transcriptional changes in a sex-dependent manner

doi: 10.1038/s41398-025-03468-3

Figure Lengend Snippet: Chd8 -inKO male : a Time spent in the center of the open field was decreased in inKO (*** p = 0.0003, t = 4.234, df = 24, two-way student’s t-test). b Time spent in the light area of the dark-light exploration test. c Time spent in the open arm of the elevated plus maze. d Time self-grooming within a 10 min time period. e Number of marbles buried in 30 min time period. (** p = 0.0022, t = 3.426, df = 24, two-way student’s t-test). f Time spent immobile in tail suspension test (**** p < 0.0001, t = 5.364, df = 28, two-way student’s t-test) g Time spent immobile in forced swim test (* p = 0.0474, t = 2.073, df = 28, two-way student’s t-test). h Motor coordination and balance were assessed using the rotarod. Time spent before falling off rotarod. i Percent freezing in each of three tones in cue fear conditioning test j Time spent in interaction zone with stranger mice and empty chamber in three-chamber social interaction test (Two way ANOVA, Stranger effect: p < 0.0001, F(1, 42) = 70.45, Tukey’s post hoc test **** p < 0.0001). n = wt 11–13 inKO 13–15. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Chd8 -inKO Female Mice : k Time spent in the center of the open field. l Time spent in the light area of the dark-light exploration test. m Time spent in the open arm of the elevated plus maze. n Time self-grooming within a 10 min time period. (**** p < 0.0001, t = 5.738, df = 23, two-way student’s t-test). o Number of marbles buried in 30 min time period. p Time spent immobile in tail suspension test q Time spent immobile in forced swim test. r Motor coordination and balance were assessed using the rotarod. Time spent before falling off rotarod. s Percent freezing in each of three tones in cue fear conditioning test t Time spent in interaction zone with stranger mice and empty chamber in three-chamber social interaction test (Two way ANOVA, Stranger effect: p < 0.0001, F(1, 48) = 40.56, Tukey’s post hoc test *** p = 0.001, *** p = 0.0006). n = wt10-13 inKO13–15. *** p < 0.001, **** p < 0.0001.

Article Snippet: To induce conditional knockdown of Chd8 in excitatory neurons, the CamKiia-Cre-Ert C57BL6 mouse line (kindly provided by the laboratory of Prof. Alon Chen, Weizmann Institute of Science, Israel) was crossed with Chd8 lx/flx C57BL6 mice (The Jackson Laboratory, Strain #:031555) to generate the Chd8 lx/flx Camk2aCreErt +/− cKO mice.

Techniques: Suspension

Journal: Translational Psychiatry

Article Title: CHD8 adulthood microglial knockdown in C57BL6 mice induces behavioral, morphological, and transcriptional changes in a sex-dependent manner

doi: 10.1038/s41398-025-03468-3

Figure Lengend Snippet: Microglial morphology was assessed in the hippocampus and prefrontal cortex (PFC) of male and female Chd8 -imKO mice. a Representative pictures of IBA-1 positive cells in imKO and wt mice in male and female PFC. b Representative pictures of IBA-1 positive cells in imKO and wt mice in male and female hippocampus. c Microglia number (IBA-1+ cells) in hippocampus of male mice (** p = 0.0070, t = 2.868, df = 34, n = wt-17 imKO-19 two-way student’s t test) d IBA-1 positive cells in PFC of male mice (*** p = 0.0007, t = 4.128, df = 17, n = wt-11 imKO-8, two-way student’s t test). e Sholl analysis (two-way ANOVA, genotype effect, **** p < 0.0001, F(1, 3212) = 144.1) and f dendrite length of microglial cells in the PFC indicates a reduction in cell complexity (*** p = 0.0001, t = 3.990, df = 95, two-way student’s t-test). g IBA-1+ cells in hippocampus of female mice ( n = 15 wild type, 15 imKO). h IBA-1+ cells in PFC of female mice ( n = 11 wild type, 15 imKO). i Sholl analysis in the PFC of female mice j Measurement of microglial dendrite length in the PFC of female mice. Representative Tracking of microglia processes in k – m WT, male imKO, and female imKO mice using Imaris. ** p < 0.01, *** p < 0.001.

Article Snippet: To induce conditional knockdown of Chd8 in excitatory neurons, the CamKiia-Cre-Ert C57BL6 mouse line (kindly provided by the laboratory of Prof. Alon Chen, Weizmann Institute of Science, Israel) was crossed with Chd8 lx/flx C57BL6 mice (The Jackson Laboratory, Strain #:031555) to generate the Chd8 lx/flx Camk2aCreErt +/− cKO mice.

Techniques:

Journal: Translational Psychiatry

Article Title: CHD8 adulthood microglial knockdown in C57BL6 mice induces behavioral, morphological, and transcriptional changes in a sex-dependent manner

doi: 10.1038/s41398-025-03468-3

Figure Lengend Snippet: RNA-Seq analysis was performed on microglia from the whole brain tissues obtained from male and female Chd8 -imKO mice. a Microglia sorting from the whole brain and b FACS analysis by three antibodies (CD45+, TMEM119+, Cd11b+) that together identify specifically microglia. The presorted CD45+ population is presented. TMEM119 is represented on the Y axis, and CD11b on the X axis. Blue cells are TMEM+CD11b+, red cells are TMEM-CD11b-, and purple cells are TMEM+CD11b-. c, d Volcano plot illustrating differentially expressed genes in the microglia of c male and d female Chd8 -inKO mice. Male microglia displayed 1381 differentially expressed genes (Of those, 773 were upregulated in the imKO microglia and 608 were downregulated) and female microglia displayed 75 differentially expressed genes (Of those, 18 were upregulated in the imKO microglia and 57 were downregulated. e, f Gene ontology of differentially expressed genes in e male and f female imKO mice. Principle Component Analysis showing distribution of all samples in this analysis ( n = 4–5 per group) found in Supplementary Fig. .

Article Snippet: To induce conditional knockdown of Chd8 in excitatory neurons, the CamKiia-Cre-Ert C57BL6 mouse line (kindly provided by the laboratory of Prof. Alon Chen, Weizmann Institute of Science, Israel) was crossed with Chd8 lx/flx C57BL6 mice (The Jackson Laboratory, Strain #:031555) to generate the Chd8 lx/flx Camk2aCreErt +/− cKO mice.

Techniques: RNA Sequencing

Journal: Translational Psychiatry

Article Title: CHD8 adulthood microglial knockdown in C57BL6 mice induces behavioral, morphological, and transcriptional changes in a sex-dependent manner

doi: 10.1038/s41398-025-03468-3

Figure Lengend Snippet: RNA-Seq analysis of hippocampal samples from Chd8 -inKO male and female mice. Volcano plots highlight minor transcriptional alterations in Chd8 -inKO a males and b females. c, d Gene Set Enrichment Analysis (GSEA) detects enrichment for genes within the Hedgehog and Wnt/Beta-catenin pathways that are upregulated specifically in males c but not changed in females d .

Article Snippet: To induce conditional knockdown of Chd8 in excitatory neurons, the CamKiia-Cre-Ert C57BL6 mouse line (kindly provided by the laboratory of Prof. Alon Chen, Weizmann Institute of Science, Israel) was crossed with Chd8 lx/flx C57BL6 mice (The Jackson Laboratory, Strain #:031555) to generate the Chd8 lx/flx Camk2aCreErt +/− cKO mice.

Techniques: RNA Sequencing

Journal: Translational Psychiatry

Article Title: CHD8 adulthood microglial knockdown in C57BL6 mice induces behavioral, morphological, and transcriptional changes in a sex-dependent manner

doi: 10.1038/s41398-025-03468-3

Figure Lengend Snippet: a Timeline of tamoxifen administration and animal sacrifice of two animal models for immunohistochemistry analysis. b, c Immunohistochemistry of the infralimbic cortex with antibodies targeting microglia (IbaI positive) and CHD8 in male b and female c mice. Nuclei are stained in blue (Hoechst). d, e Immunohistochemistry of hippocampus with antibodies targeting neurons (NeuN), and CHD8 in male d and female e mice. Nuclei are stained in blue (Hoechst).

Article Snippet: These mice were crossed with Chd8 flx/flx mice (The Jackson Laboratory, Strain #:031555) to generate the Chd8 flx/flx Cx3cr1CreErt +/− (cKO) and Chd8 flx/flx Cx3cr1CreErt −/− (wild-type) mice.

Techniques: Immunohistochemistry, Staining

Journal: Translational Psychiatry

Article Title: CHD8 adulthood microglial knockdown in C57BL6 mice induces behavioral, morphological, and transcriptional changes in a sex-dependent manner

doi: 10.1038/s41398-025-03468-3

Figure Lengend Snippet: Chd8 -imKO males : a Time spent in the center of the open field. (* p = 0.0205, t = 2.554, df = 17, two-way student’s t-test) b Time spent in the light area of the dark-light exploration test. (* p = 0.0473, t = 2.149, df = 16, two-way student’s t-test) c Time spent in the open arm of the elevated plus maze (* p = 0.0242, t = 2.527, df = 14, two-way student’s t-test). d Time self-grooming within a 10 min time period. e Number of marbles buried in 30 min time period. f Time spent immobile in tail suspension test (* p = 0.0195, t = 2.580, df = 17, two-way student’s t-test) g Time spent immobile in forced swim test (** p = 0.0037, t = 3.437, df = 15, two-way student’s t-test). h Motor coordination and balance were assessed using the rotarod. Time spent before falling off rotarod. i Percent freezing in each of three tones in cue fear conditioning test (Two way ANOVA, Genotype effect: p = 0.0055, F(1, 17) = 10.09, Tukey’s post hoc test, * p = 0.0172, ** p = 0.0016, * p = 0.0173). j Time spent in interaction zone with stranger mice and empty chamber in three-chamber social interaction test (Two way ANOVA, Stranger effect: p < 0.0001, F(1, 62) = 22.68, Tukey’s post hoc test, *** p < 0.0001, ns p = 0.1589). n = wt 9–11 imKO 7–8. * p < 0.05, ** p < 0.01, *** p < 0.001. Chd8 -imKO females : k Time spent in the center of the open field (* p = 0.0239, t = 2.479, df = 17, two-way student’s t-test). l Time spent in the light area of the dark-light exploration test. m Time spent in the open arm of the elevated plus maze. n Time self-grooming within a 10 min time period. o Number of marbles buried in 30 min time period. p Time spent immobile in tail suspension test q Time spent immobile in forced swim test. r Motor coordination and balance were assessed using the rotarod. Time spent before falling off rotarod. s Percent freezing in each of three tones in cue fear conditioning test t Time spent in interaction zone with stranger mice and empty chamber in three-chamber social interaction test (Two way ANOVA, Stranger effect: p < 0.0001, F(1, 34) = 24.59, Tukey’s post hoc test * p = 0.0169, ** p = 0.0027). Further tests determined no differences between experimental groups. (l- dark-light, m- elevated plus maze, p- tail suspension test, q- forced swimming test). n = wt 9–10 imKO 9–10. * p < 0.05, ** p < 0.01.

Article Snippet: These mice were crossed with Chd8 flx/flx mice (The Jackson Laboratory, Strain #:031555) to generate the Chd8 flx/flx Cx3cr1CreErt +/− (cKO) and Chd8 flx/flx Cx3cr1CreErt −/− (wild-type) mice.

Techniques: Suspension

Journal: Translational Psychiatry

Article Title: CHD8 adulthood microglial knockdown in C57BL6 mice induces behavioral, morphological, and transcriptional changes in a sex-dependent manner

doi: 10.1038/s41398-025-03468-3

Figure Lengend Snippet: Chd8 -inKO male : a Time spent in the center of the open field was decreased in inKO (*** p = 0.0003, t = 4.234, df = 24, two-way student’s t-test). b Time spent in the light area of the dark-light exploration test. c Time spent in the open arm of the elevated plus maze. d Time self-grooming within a 10 min time period. e Number of marbles buried in 30 min time period. (** p = 0.0022, t = 3.426, df = 24, two-way student’s t-test). f Time spent immobile in tail suspension test (**** p < 0.0001, t = 5.364, df = 28, two-way student’s t-test) g Time spent immobile in forced swim test (* p = 0.0474, t = 2.073, df = 28, two-way student’s t-test). h Motor coordination and balance were assessed using the rotarod. Time spent before falling off rotarod. i Percent freezing in each of three tones in cue fear conditioning test j Time spent in interaction zone with stranger mice and empty chamber in three-chamber social interaction test (Two way ANOVA, Stranger effect: p < 0.0001, F(1, 42) = 70.45, Tukey’s post hoc test **** p < 0.0001). n = wt 11–13 inKO 13–15. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Chd8 -inKO Female Mice : k Time spent in the center of the open field. l Time spent in the light area of the dark-light exploration test. m Time spent in the open arm of the elevated plus maze. n Time self-grooming within a 10 min time period. (**** p < 0.0001, t = 5.738, df = 23, two-way student’s t-test). o Number of marbles buried in 30 min time period. p Time spent immobile in tail suspension test q Time spent immobile in forced swim test. r Motor coordination and balance were assessed using the rotarod. Time spent before falling off rotarod. s Percent freezing in each of three tones in cue fear conditioning test t Time spent in interaction zone with stranger mice and empty chamber in three-chamber social interaction test (Two way ANOVA, Stranger effect: p < 0.0001, F(1, 48) = 40.56, Tukey’s post hoc test *** p = 0.001, *** p = 0.0006). n = wt10-13 inKO13–15. *** p < 0.001, **** p < 0.0001.

Article Snippet: These mice were crossed with Chd8 flx/flx mice (The Jackson Laboratory, Strain #:031555) to generate the Chd8 flx/flx Cx3cr1CreErt +/− (cKO) and Chd8 flx/flx Cx3cr1CreErt −/− (wild-type) mice.

Techniques: Suspension

Journal: Translational Psychiatry

Article Title: CHD8 adulthood microglial knockdown in C57BL6 mice induces behavioral, morphological, and transcriptional changes in a sex-dependent manner

doi: 10.1038/s41398-025-03468-3

Figure Lengend Snippet: Microglial morphology was assessed in the hippocampus and prefrontal cortex (PFC) of male and female Chd8 -imKO mice. a Representative pictures of IBA-1 positive cells in imKO and wt mice in male and female PFC. b Representative pictures of IBA-1 positive cells in imKO and wt mice in male and female hippocampus. c Microglia number (IBA-1+ cells) in hippocampus of male mice (** p = 0.0070, t = 2.868, df = 34, n = wt-17 imKO-19 two-way student’s t test) d IBA-1 positive cells in PFC of male mice (*** p = 0.0007, t = 4.128, df = 17, n = wt-11 imKO-8, two-way student’s t test). e Sholl analysis (two-way ANOVA, genotype effect, **** p < 0.0001, F(1, 3212) = 144.1) and f dendrite length of microglial cells in the PFC indicates a reduction in cell complexity (*** p = 0.0001, t = 3.990, df = 95, two-way student’s t-test). g IBA-1+ cells in hippocampus of female mice ( n = 15 wild type, 15 imKO). h IBA-1+ cells in PFC of female mice ( n = 11 wild type, 15 imKO). i Sholl analysis in the PFC of female mice j Measurement of microglial dendrite length in the PFC of female mice. Representative Tracking of microglia processes in k – m WT, male imKO, and female imKO mice using Imaris. ** p < 0.01, *** p < 0.001.

Article Snippet: These mice were crossed with Chd8 flx/flx mice (The Jackson Laboratory, Strain #:031555) to generate the Chd8 flx/flx Cx3cr1CreErt +/− (cKO) and Chd8 flx/flx Cx3cr1CreErt −/− (wild-type) mice.

Techniques:

Journal: Translational Psychiatry

Article Title: CHD8 adulthood microglial knockdown in C57BL6 mice induces behavioral, morphological, and transcriptional changes in a sex-dependent manner

doi: 10.1038/s41398-025-03468-3

Figure Lengend Snippet: RNA-Seq analysis was performed on microglia from the whole brain tissues obtained from male and female Chd8 -imKO mice. a Microglia sorting from the whole brain and b FACS analysis by three antibodies (CD45+, TMEM119+, Cd11b+) that together identify specifically microglia. The presorted CD45+ population is presented. TMEM119 is represented on the Y axis, and CD11b on the X axis. Blue cells are TMEM+CD11b+, red cells are TMEM-CD11b-, and purple cells are TMEM+CD11b-. c, d Volcano plot illustrating differentially expressed genes in the microglia of c male and d female Chd8 -inKO mice. Male microglia displayed 1381 differentially expressed genes (Of those, 773 were upregulated in the imKO microglia and 608 were downregulated) and female microglia displayed 75 differentially expressed genes (Of those, 18 were upregulated in the imKO microglia and 57 were downregulated. e, f Gene ontology of differentially expressed genes in e male and f female imKO mice. Principle Component Analysis showing distribution of all samples in this analysis ( n = 4–5 per group) found in Supplementary Fig. .

Article Snippet: These mice were crossed with Chd8 flx/flx mice (The Jackson Laboratory, Strain #:031555) to generate the Chd8 flx/flx Cx3cr1CreErt +/− (cKO) and Chd8 flx/flx Cx3cr1CreErt −/− (wild-type) mice.

Techniques: RNA Sequencing

Journal: Translational Psychiatry

Article Title: CHD8 adulthood microglial knockdown in C57BL6 mice induces behavioral, morphological, and transcriptional changes in a sex-dependent manner

doi: 10.1038/s41398-025-03468-3

Figure Lengend Snippet: RNA-Seq analysis of hippocampal samples from Chd8 -inKO male and female mice. Volcano plots highlight minor transcriptional alterations in Chd8 -inKO a males and b females. c, d Gene Set Enrichment Analysis (GSEA) detects enrichment for genes within the Hedgehog and Wnt/Beta-catenin pathways that are upregulated specifically in males c but not changed in females d .

Article Snippet: These mice were crossed with Chd8 flx/flx mice (The Jackson Laboratory, Strain #:031555) to generate the Chd8 flx/flx Cx3cr1CreErt +/− (cKO) and Chd8 flx/flx Cx3cr1CreErt −/− (wild-type) mice.

Techniques: RNA Sequencing