Journal: bioRxiv

Article Title: Cognitive dysfunction following brain trauma results from sex-specific reactivation of the developmental pruning processes

doi: 10.1101/2024.08.13.607610

Figure Lengend Snippet: (A, D, G, J) Schematic illustrations in the non-injured quadpartite hippocampal synapse and at 1 dpi of SRR fl/fl and TMEM119 creERT2 :SRR fl/fl mice (created with Biorender). (B-D) Western blot of whole hippocampus shows a significant increase in (B) GluN2A and (C) GluN2B expression at 1 dpi in SRR fl/fl male mice that is not present in microglial knockout mice. (E-G) Expression of (E) phosphorylated GluN2B and (F) DAPK1 is increased at 1 dpi in SRR fl/fl mice and not until 3 dpi in TMEM119 creERT2 :SRR fl/fl mice. (H,J) The ratio of phosphorylated to non-phosphorylated CREB is significantly upregulated in TMEM119 creERT2 :SRR fl/fl but not SRR fl/fl male mice after injury. (I,J) BDNF is significantly increased at 1 and 3 dpi in TMEM119 creERT2 :SRR fl/fl male mice, prior to upregulation in SRR fl/fl mice at 3 dpi. (K) GluN2B is downregulated in isolated hippocampal membrane fractions in male and female WT mice at 3 dpi, however (L) the ratio of phosphorylated GluN2B is increased in male but decreased in female mice at 3 dpi. *p<0.05, **p<0.01, ***p<0.001. Two-way ANOVA with Tukey’s multiple comparison test; n=4-5. Values represent mean ± SEM

Article Snippet: Total protein transferred was imaged and membranes were blocked for 1 hour at RT with 5% bovine serum albumin (BSA) in 1X PBS followed by overnight incubation at 4°C with primary antibodies diluted in 5% BSA (1:1000 unless otherwise noted): rabbit anti-BDNF (Abcam, Boston, MA, #ab9793), rabbit anti-phospho-ERK1/2 (1:500; Cell Signaling, Danvers, MA, #9101S), rabbit anti-ERK1/2 (Cell Signaling #9102), rabbit anti-NR1 (Millipore #05-432), rabbit anti-NMDAR1 (Abcam #ab109182), rabbit anti-NR2A (Millipore #07-632), rabbit anti-NR2B (Millipore #06-600), rabbit anti-GRIN2B (1:500; ThermoFisher, #21920-1-AP), rabbit anti-DAPK1 (Cell Signaling, Danvers, MA, #3008S), Rabbit anti-p-CREB (ABclonal, Woburn, MA, #AP0019), rabbit anti-CREB (ABclonal, #A11064), rabbit anti-Sodium Potassium ATPase (1:500; Abcam, #AB76020), rabbit anti-Phospho-N2B,Ser1303 (Millipore, #07-398), rabbit anti-C1qA (Abcam, #ab189922), rabbit anti-C3 (Invitrogen, #PA5-21349), mouse anti-GPR56 (Biolegend, #358205), mouse anti-SRR (Santa Cruz Biotech, Dallas, TX, #sc-365217), rabbit anti-Slc1a4 (Cell Signaling, #8442S), and mouse anti-GAPDH (1:5000, Santa Cruz Biotech, #47724).

Techniques: Western Blot, Expressing, Knock-Out, Isolation, Membrane, Comparison

Journal: iScience

Article Title: A cingulate-hippocampal circuit mediates early depressive-like behavior in the mouse model of Alzheimer disease

doi: 10.1016/j.isci.2024.109778

Figure Lengend Snippet: Nrg1 overexpression in the ACC induces synaptic deficit and depressive-like behaviors (A) Schematic showing the injections of overexpressed virus rAAV2/9-CamkⅡα-Nrg1-mCherry-WPREs-pA (or control virus rAAV2/9-CamkⅡα-mCherry-WPREs-pA) into the ACC of C57BL/6J mice bilaterally. Fluorescent images showing neurons in ACC are infected with overexpressed virus (AAV-OENrg1). Scale bar, 100 μm. (B) Representative western blots (left panel) and quantification (right panel) of Nrg1 protein levels of mice injected with AAV-OENrg1 and AAV-mCherry. Expression level of Nrg1 is significantly increased after AAV-OENrg1 injection compared with AAV-mCherry group. Student’s t test. n = 4 mice per group. (C) Representative Golgi staining of dendrites in ACC. Scale bar, 20 μm. (D) Sholl analysis of apical (left panel) and basal dendrites (right panel) of ACC pyramidal neurons from AAV-mCherry and AAV-OENrg1 mice. (E) Representative images of spines density and labeled spines of apical and basal dendrites. Nrg1 overexpression decreases both apical node and spines (left panel) and basal node and spines (right panel) in ACC. Scale bar, 10 μm. Student’s t test. n = 36 neurons from six mice per group. (F) Representative western blots (left panel) and quantification (right panel) of Nrg1, GluR1, GluR2, NR2A, NR2B, and PAD95 protein levels of mice injected with AAV-OENrg1 and AAV-mCherry. The expression levels of the AMPA receptor (AMPAR) subunits (GluR1, GluR2), NMDA receptor (NMDAR) subunits (NR2A, NR2B), and PSD95 are decreased after AAV-OENrg1 injection into the ACC compared with AAV-mCherry group. Student’s t test. n = 4 mice per group. (G and H) Representative traces (G) and quantification (H) of mEPSCs in the ACC region of AAV-mCherry and AAV-OENrg1 mice. AAV-OENrg1 mice show significantly decreased frequency and amplitude of mEPSCs compared with the AAV-mCherry mice. Student’s t test. n = 10 neurons from three mice per group. (I) Compared to control mice, AAV-OENrg1 mice displayed increased immobility time in the TST (left panel) and FST (right panel). Student’s t test. n = 10 mice in AAV-mCherry group and 13 mice in AAV-OENrg1 group. (J) Compared to control mice, AAV-OENrg1 mice displayed decreased sucrose preference score (%) (left panel) and sucrose consumption (g) (right panel) in the SPT. Student’s t test. Data represent mean ± SEM; ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Rabbit monoclonal anti-NR2A , Abcam , Cat# ab124913 RRID: AB_10975154.

Techniques: Over Expression, Virus, Infection, Western Blot, Injection, Expressing, Staining, Labeling

Journal: iScience

Article Title: A cingulate-hippocampal circuit mediates early depressive-like behavior in the mouse model of Alzheimer disease

doi: 10.1016/j.isci.2024.109778

Figure Lengend Snippet:

Article Snippet: Rabbit monoclonal anti-NR2A , Abcam , Cat# ab124913 RRID: AB_10975154.

Techniques: Virus, Recombinant, Software