Journal: bioRxiv

Article Title: Persistent cortical excitatory neuron dysregulation in adult Chd8 haploinsufficient mice

doi: 10.1101/2025.06.04.657776

Figure Lengend Snippet: A) Barplot of DEG counts across bulk RNA-seq experiments (constitutive Chd8 +/- , Camk2a-CRE conditional Chd8 +/flox , and nuclear and synaptosomal preparations from the Chd8 +/- line. B) Chd8 is down-regulated in all but the nuclear preparation. C) LogFC concordance between constitutive and conditional datasets for constitutive model DEGs. D) LogFC concordance plots for nuclear and synaptosomal versus whole cortex for DEGs identified in nulear or synaptosomal fractions on left, reduced concordance between nuclear and synapsomal DEG logFC on right. E) Selected enriched GO terms for upregulated (left) and downregulated (right) genes across experiments. F) Downregulation of Dlg2 in synaptosomal RNA. GH) Western blot validating decreased synaptosomal PSD93 protein in Chd8 +/- cortex.

Article Snippet: Primary antibody PSD93 (1:50, DSHB, Clone N18/28, supernatant fraction) was diluted in 7.5 ml Intercept PBS blocking buffer with 0.1% Tween.

Techniques: RNA Sequencing, Western Blot

Journal: bioRxiv

Article Title: Adult medial habenula neurons require GDNF receptor GFRα1 for synaptic stability and function

doi: 10.1101/2021.07.07.451421

Figure Lengend Snippet: A) dTomato epifluorescence (red) in sagittal sections of Gfra1 dTOM mouse brain injected with tamoxifen at 3 months counterstained with DAPI (blue). FR: fasciculus retroflexus; Hipp: hippocampus; HTH: hypothalamus; IPN: interpeduncular nucleus; LS: lateral septum; mHb: medial habenula; MS; medial septum; Str: striatum. Scale bar, 500μm. B) GFP (green) and ChAT (red) immunostaining in sagittal sections of 3 month old Gfra1 GFP mouse brain counterstained with DAPI (blue). 3V: 3th ventricle; LHb: lateral habenula. Scale bar, 400μm. C-D) GFP (green), ChAT (red) and SP (grey) immunostaining in coronal sections of the mHb (C) and IPN (D) of 3 month old Gfra1 GFP mouse brain. dmHb: dorsal medial habenula; vmHb: ventral medial habenula: dIPN: dorsal IPN, vIPN: ventral IPN; IPL: lateral IPN. Scale bars, 200μm. E) GFRα1 immunostaining (red) in coronal sections of the mHb and IPN from 3 month old control (C57BL6/J) mouse. Scale bars, 200μm. F) GFP (green) and GFRα1 (red) immunostaining in coronal sections through the IPN of a 3 month old Gad67 Cre ;R1CG fx/fx mouse. Scale bars, 100μm (merged) and 35μm (inset). G) GFP (green) and calretinin (CR, red) immunostaining in coronal sections of the BAC and TS from 3 month old Gfra1 GFP mouse brain counterstained with DAPI (blue). Scale bars, 100μm (BAC) and 200μm (TS). H,I) Immunoblots of whole (We), cytosolic (Ct) and synaptosome (Sn) protein extracts from the mHb and IPN of 3 month old C57BL6/J mice probed for GFRα1 and NCAM (H) or RET, GDNF and PSD95 (I). Tubulin was probed as loading control. J, L) GFRα1 (green) and PSD93 (red) immunostaining in the mHb (J) and IPN (L) of 3 month old C57BL6/J mice. Scale bars, 10μm (merged) and 5μm (inset). K, M) Quantification (±SEM) of GFRα1, PSD93 and double-labeled puncta in mHb (K) and IPN (M). N=5 mice (25-30 images per mouse).

Article Snippet: The primary antibodies used were guinea pig anti VGlut1 (1:500, AB2251, Millipore); Mouse anti VGlut2 (1:500, 135421, SYSY); rabbit anti PSD93 (1:500, 124102, SYSY); Goat anti GFRα1 (1:500, AF560, RnD); guinea pig anti VGAT (1:500, 131004, SYSY); Mouse anti gephyrin (1:500, 147111, SYSY); goat anti synapsin Ia/b (1:500, sc8295, Santa Cruz).

Techniques: Injection, Immunostaining, Western Blot, Labeling

Journal: bioRxiv

Article Title: Adult medial habenula neurons require GDNF receptor GFRα1 for synaptic stability and function

doi: 10.1101/2021.07.07.451421

Figure Lengend Snippet: A) Synapsin-I (green) and PSD93 (red) immunostaining in coronal sections of the mHb and IPN of WT, Het and KO mice. Scale bar, 10μm. B) Quantification (±SEM) of puncta with co-localized immunoreactivity for Synapsin-I and PSD93 in mHb and IPN. N=4-5 mice per group (25-30 images per mouse); 1-way ANOVA followed by Tukey’s post-hoc test; *, P <0.05. C) VGlut1 (green) immunostaining in coronal sections of the mHb and IPN of WT, Het and KO mice. Scale bar, 10μm. D) Quantification (±SEM) of VGlut1 puncta in mHb (K) and IPN in mHb and IPN. N=5 mice per group (25-30 images per mouse); 1-way ANOVA followed by Tukey’s post-hoc test; *, P <0.05; ***, P<0.001. E, G) Synapsin-I (red), PSD93 (purple), Vglut1 (green) and Vglut2 (red) immunostaining in coronal sections of the mHb (E) and IPN (G) in mHb.KO or mHb.WT mice. Scale bar, 10 μm. F, H) Quantification (±SEM) of immunoreactive puncta for each marker in mHb (E) and IPN (G) of mHb.KO or mHb.WT mice. N=6-7 mice per group (25-30 images per mouse); Student’s T-test; *, P< 0.05; ****, P< 0.0001.

Article Snippet: The primary antibodies used were guinea pig anti VGlut1 (1:500, AB2251, Millipore); Mouse anti VGlut2 (1:500, 135421, SYSY); rabbit anti PSD93 (1:500, 124102, SYSY); Goat anti GFRα1 (1:500, AF560, RnD); guinea pig anti VGAT (1:500, 131004, SYSY); Mouse anti gephyrin (1:500, 147111, SYSY); goat anti synapsin Ia/b (1:500, sc8295, Santa Cruz).

Techniques: Immunostaining, Marker