Journal: bioRxiv

Article Title: ECSIT prevents Alzheimer’s disease pathology by regulating neuronal mitochondrial ROS and mitophagy

doi: 10.1101/2020.01.09.900407

Figure Lengend Snippet: a, Mitochondrial mass assessed by qPCR. Data are expressed as fold-increase relative to WT. Average of 6 mice per group is shown. Not significant in Mann-Whitney test for each region. b, Representative immunofluorescence of GFP and stainings for ECSIT and TOM6 and confocal acquisition after infection of primary neurons with AAV-GFP and AAV-ECSIT-GFP. Lower intensity for ECSIT staining is shown on the left to distinguish better mitochondrial localization. Images processed with Fiji. c, ECSIT immunostaining and confocal imaging of coronal brain sections of mice injected with AAV-GFP-ECSIT and AAV-GFP 8 weeks after injection. Objective 10x, scale bar 500 µm. Images representative of 3 independent experiments. d, Western blot analysis of indicated proteins in lysates of cortex (Cx), hippocampus (Hpc) and Midbrain (Mb) of WT animals injected with AAV-GFP (G) or AAV-GFP-ECSIT (E). Representative blots of 2 experiments. e, Cued fear conditioning (FC) on 3- to 5-month-old WT or APP/PS1+virus animals tested in . N=12 WT+GFP; 15 WT +ECSIT; 9 APP/PS1+GFP; 11 APP/PS1+ECSIT. f, Visual, motor and motivational controls for animals tested in . g, Sensory threshold assessment (STA) of animals tested in and . h, Basal synaptic transmission in hippocampus for samples represented in .

Article Snippet: AAV-GFP (AAV9-hSYN1-eGFP-WPRE, Vector Biolabs) and custom-made AAV-ECSIT-GFP (AAV9-hSYN1-eGFP-2A-mECSIT-WPRE, with eGFP and mouse ECSIT2 expression driven by the same hSYN1 promoter, with a T2A peptide linking eGFP and ECSIT, Vector Biolabs) were injected bilaterally into the dorsal hippocampus at the following coordinates (flat skull position): antero-posterior: −1.9 mm, medio-lateral: +/-1.8mm, dorso-ventral: −1.8 mm below dural surface as calculated relative to bregma according to the stereotaxic atlas.

Techniques: MANN-WHITNEY, Immunofluorescence, Infection, Staining, Immunostaining, Imaging, Injection, Western Blot, Transmission Assay

Journal: Cells

Article Title: Selective Menin Deletion in the Hippocampal CA1 Region Leads to Disruption of Contextual Memory in the MEN1 Conditional Knockout Mouse: Behavioral Restoration and Gain of Function following the Reintroduction of MEN1 Gene.

doi: 10.3390/cells11244019

Figure Lengend Snippet: Figure 2. The transduction of hippocampal neuronal cultures from the MEN1 floxed mice with the virus, pNN.AAV.hSyn.HI.eGFP-Cre.WPRE.SV40. (A) Controls with no virus; (i) GFP signal in the absence of virus; (ii) DAPI for nuclear staining; (iii) merge with GFP and DAPI; (B) CKO with virus transduction by Cre AAV9; (i) GFP signal indicating virus transduction in the preparation with the Cre AAV9; (ii) DAPI for nuclear staining; (iii) merge with GFP and DAPI showing overlap in the neurons expressing GFP and DAPI.

Article Snippet: Ready-to-use AAV9 particles produced from pENN.AAV.hSyn.HI.eGFP-Cre.WPRE.SV40 (#105540) were ordered from Addgene.

Techniques: Transduction, Virus, Staining, Expressing

Journal: Cells

Article Title: Selective Menin Deletion in the Hippocampal CA1 Region Leads to Disruption of Contextual Memory in the MEN1 Conditional Knockout Mouse: Behavioral Restoration and Gain of Function following the Reintroduction of MEN1 Gene.

doi: 10.3390/cells11244019

Figure Lengend Snippet: Figure 3. In vitro virus transduction of the hippocampal neurons from MEN1 floxed mice in control and CKO groups. (A) Controls with no virus; (i) the expression of DAP for nuclear staining; (ii) GFP signal in the absence of virus; (iii) C-menin showing axoplasmic expression; (iv) merge in controls for GFP and DAPI; (B) CKOs with virus; (i) DAPI for nuclear staining; (ii) GFP signal indicating virus transduction in the preparation with the Cre AAV9; (iii) C-menin signal weaker compared to the controls; (iv) merge in the CKO for GFP and DAPI; (C) summary data, showing the reduction in menin signals as measured by mean gray-to-area ratio in the CKO compared to the controls. Data represent the mean ratio ± SEM. n = 4. Statistical significance (Mann–Whitney U test), * p < 0.05.

Article Snippet: Ready-to-use AAV9 particles produced from pENN.AAV.hSyn.HI.eGFP-Cre.WPRE.SV40 (#105540) were ordered from Addgene.

Techniques: In Vitro, Virus, Transduction, Control, Expressing, Staining, MANN-WHITNEY

Journal: Cells

Article Title: Selective Menin Deletion in the Hippocampal CA1 Region Leads to Disruption of Contextual Memory in the MEN1 Conditional Knockout Mouse: Behavioral Restoration and Gain of Function following the Reintroduction of MEN1 Gene.

doi: 10.3390/cells11244019

Figure Lengend Snippet: Figure 4. In vivo virus transduction of the hippocampal CA1 from MEN1 floxed mice in CA1 region of hippocampus and dentate gyrus (DG). (A) Expression of GFP in CA1 in the CKO; (i) the expression of DAPI for nuclear staining in CA1; (ii) the expression of GFP as the marker of virus transduction in the neurons transduced by the AAV9 under the action of synapsin promoter in CA1; (iii) merge in controls with control virus AAV9 GFP and overlap between DAPI and GFP; (B) non-GFP expressing region in dentate gyrus (DG); (i) the expression of DAPI for nuclear staining in DG; (ii) GFP signal in the non-transduced region of hippocampus; (iii) merge for showing the overlap between DAPI and GFP.

Article Snippet: Ready-to-use AAV9 particles produced from pENN.AAV.hSyn.HI.eGFP-Cre.WPRE.SV40 (#105540) were ordered from Addgene.

Techniques: In Vivo, Virus, Transduction, Expressing, Staining, Marker, Control

Journal: Cells

Article Title: Selective Menin Deletion in the Hippocampal CA1 Region Leads to Disruption of Contextual Memory in the MEN1 Conditional Knockout Mouse: Behavioral Restoration and Gain of Function following the Reintroduction of MEN1 Gene.

doi: 10.3390/cells11244019

Figure Lengend Snippet: Figure 6. Ectopic expression in the hippocampus of the mice injected with AAV9 under the function of synapsin promoter and GFP as the marker for the virus transduction. (i) The expression of DAPI for nuclear staining; (ii) GFP as the marker of virus transduction in the neurons transduced by the control AAV9 under the action of synapsin promoter; (iii) merge in controls showing overlap between DAPI and GFP.

Article Snippet: Ready-to-use AAV9 particles produced from pENN.AAV.hSyn.HI.eGFP-Cre.WPRE.SV40 (#105540) were ordered from Addgene.

Techniques: Expressing, Injection, Marker, Virus, Transduction, Staining, Control