Journal: Psychiatry Investigation

Article Title: Memory Decline and Aberration of Synaptic Proteins in X-Linked Moesin Knockout Male Mice

doi: 10.30773/pi.2024.0186

Figure Lengend Snippet: Moesin-KO neurons decreased cell viability and the expression of neuronal cell markers. Relative expression levels of GFAP, MAP2, Sox2, DCX, SNAP25, PSD95, and moesin are displayed. At 7 days (A) and 14 days (B) after seeding cortical neurons, molecular changes in neuronal cell markers were examined by immunoblotting. All data were analyzed by t-test and are presented as the mean±SEM of three replicates. The bar is 100 μm. *p<0.05; **p<0.01; ***p<0.001. KO, knockout mice; WT, wild-type mice.

Article Snippet: The tissue lysates (30 μg) were electrophoresed and transferred to 0.45 μm nitrocellulose membranes, which were incubated with anti-GFAP (ab7260, 1:1,000, Abcam, Cambridge, MA, USA), anti-MAP2 (ab5392, 1:1,000, Abcam), anti-Sox2 (#4900, CST, 1:1,000), anti-DCX (#91954, CST, 1:1,000), anti-soluble NSF adapter protein 25 (SNAP25) (#5308, CST, 1:1,000), anti-postsynaptic density protein 95 (PSD95) (#3409, CST, 1:1,000), anti-phospho-PSD95 (#45737, CST, 1:1,000), or anti-moesin (#3146, CST, 1:1,000), anti-syntaxin1A (#18572, CST, 1:1,000), anti-synaptophysin (#5467, CST, 1:1,000), anti-CDK5 (#14145, CST, 1:1,000), anti-radixin (#2636, CST, 1:1,000), anti-synapsin 1 (#2312, CST, 1:1,000), anti-phospho-synapsin 1 (Ser9) (#2311, CST, 1:1,000), anti-phospho-synapsin 1 (Ser605) (#88246, CST, 1:1,000), anti-phospho-synapsin 1 (Ser549) (#PA1-4697, 1:1,000; Thermo Fisher Scientific, Waltham, MA, USA), anti-ERK (#4695, CST, 1:1,000), anti-phospho-ERK (#4370, CST, 1:1,000), anti-CREB (#9197, CST, 1:1,000), anti-phospho-CREB (#9190, CST, 1:1,000), anti-mammalian uncoordinated18 (MUNC18) (#13414, CST, 1:1,000), anti-phospho-MUNC18 (ab183484, 1:1,000, Abcam) anti-β-actin (#4967, CST, 1:1,000) for 16 hours at 4°C.

Techniques: Expressing, Western Blot, Knock-Out

Journal: Psychiatry Investigation

Article Title: Memory Decline and Aberration of Synaptic Proteins in X-Linked Moesin Knockout Male Mice

doi: 10.30773/pi.2024.0186

Figure Lengend Snippet: Moesin-KO decreased the expression of synaptic proteins and phosphorylation of signaling proteins in the adult brain. A and B: The expression or activation level of synaptic vesicle proteins such as Syntaxin 1A, Synaptophysin, CDK5, and radixin were confirmed by immunoblotting in the cortex and hippocampus to examine the downstream target of moesin. Relative expression levels of each protein are presented in bar graphs. C and D: The expression and phosphorylation level of synapsin I were examined in the cortex and hippocampus of moesin-KO and presented in bar graphs. E and F: The activation levels of signaling molecules such as ERK and CREB were checked with phosphorylationspecific antibodies. The expression and phosphorylation levels of MUNC18 and PSD95 were examined in the cortex, and hippocampus of moesin-KO, and their relative expression is presented in graphs. All data were analyzed by t-test and are presented as the mean±SEM of three replicates. **p<0.01; ***p<0.001. KO, knockout mice; WT, wild-type mice; CDK5, Cyclin dependent kinase 5; ERK, extracellular signal-regulated kinase; CREB, cAMP response element-binding protein; SEM, standard error of the mean.

Article Snippet: The tissue lysates (30 μg) were electrophoresed and transferred to 0.45 μm nitrocellulose membranes, which were incubated with anti-GFAP (ab7260, 1:1,000, Abcam, Cambridge, MA, USA), anti-MAP2 (ab5392, 1:1,000, Abcam), anti-Sox2 (#4900, CST, 1:1,000), anti-DCX (#91954, CST, 1:1,000), anti-soluble NSF adapter protein 25 (SNAP25) (#5308, CST, 1:1,000), anti-postsynaptic density protein 95 (PSD95) (#3409, CST, 1:1,000), anti-phospho-PSD95 (#45737, CST, 1:1,000), or anti-moesin (#3146, CST, 1:1,000), anti-syntaxin1A (#18572, CST, 1:1,000), anti-synaptophysin (#5467, CST, 1:1,000), anti-CDK5 (#14145, CST, 1:1,000), anti-radixin (#2636, CST, 1:1,000), anti-synapsin 1 (#2312, CST, 1:1,000), anti-phospho-synapsin 1 (Ser9) (#2311, CST, 1:1,000), anti-phospho-synapsin 1 (Ser605) (#88246, CST, 1:1,000), anti-phospho-synapsin 1 (Ser549) (#PA1-4697, 1:1,000; Thermo Fisher Scientific, Waltham, MA, USA), anti-ERK (#4695, CST, 1:1,000), anti-phospho-ERK (#4370, CST, 1:1,000), anti-CREB (#9197, CST, 1:1,000), anti-phospho-CREB (#9190, CST, 1:1,000), anti-mammalian uncoordinated18 (MUNC18) (#13414, CST, 1:1,000), anti-phospho-MUNC18 (ab183484, 1:1,000, Abcam) anti-β-actin (#4967, CST, 1:1,000) for 16 hours at 4°C.

Techniques: Expressing, Phospho-proteomics, Activation Assay, Western Blot, Knock-Out, Binding Assay

Journal: Psychiatry Investigation

Article Title: Memory Decline and Aberration of Synaptic Proteins in X-Linked Moesin Knockout Male Mice

doi: 10.30773/pi.2024.0186

Figure Lengend Snippet: Risperidone reversed behavioral and molecular deficits in moesin-KO. Risperidone was administered for 2 weeks and validated by behavioral experiments at 2 months of age. A: Risperidone reversed memory impairment in moesin-KO in the passive avoidance test and (B) total entry number (left panel) and spontaneous alternation percentage (right panel) in the Y maze. C: Risperidone induced the expression of PSD95, the phosphorylation of synapsin at Ser605 and Ser549, and ERK phosphorylation in the cortex of moesin-KO, and the data are displayed in the graph. All data were analyzed by one-way ANOVA and are presented as the mean±SEM. *p<0.05; **p<0.01; ***p<0.001. KO, knockout mice; WT, wild-type mice; ERK, extracellular signal-regulated kinase; SEM, standard error of the mean; Ris, risperidone.

Article Snippet: The tissue lysates (30 μg) were electrophoresed and transferred to 0.45 μm nitrocellulose membranes, which were incubated with anti-GFAP (ab7260, 1:1,000, Abcam, Cambridge, MA, USA), anti-MAP2 (ab5392, 1:1,000, Abcam), anti-Sox2 (#4900, CST, 1:1,000), anti-DCX (#91954, CST, 1:1,000), anti-soluble NSF adapter protein 25 (SNAP25) (#5308, CST, 1:1,000), anti-postsynaptic density protein 95 (PSD95) (#3409, CST, 1:1,000), anti-phospho-PSD95 (#45737, CST, 1:1,000), or anti-moesin (#3146, CST, 1:1,000), anti-syntaxin1A (#18572, CST, 1:1,000), anti-synaptophysin (#5467, CST, 1:1,000), anti-CDK5 (#14145, CST, 1:1,000), anti-radixin (#2636, CST, 1:1,000), anti-synapsin 1 (#2312, CST, 1:1,000), anti-phospho-synapsin 1 (Ser9) (#2311, CST, 1:1,000), anti-phospho-synapsin 1 (Ser605) (#88246, CST, 1:1,000), anti-phospho-synapsin 1 (Ser549) (#PA1-4697, 1:1,000; Thermo Fisher Scientific, Waltham, MA, USA), anti-ERK (#4695, CST, 1:1,000), anti-phospho-ERK (#4370, CST, 1:1,000), anti-CREB (#9197, CST, 1:1,000), anti-phospho-CREB (#9190, CST, 1:1,000), anti-mammalian uncoordinated18 (MUNC18) (#13414, CST, 1:1,000), anti-phospho-MUNC18 (ab183484, 1:1,000, Abcam) anti-β-actin (#4967, CST, 1:1,000) for 16 hours at 4°C.

Techniques: Expressing, Phospho-proteomics, Knock-Out

Journal: International immunopharmacology

Article Title: Minocycline ameliorates cognitive impairment in rats with trigeminal neuralgia by regulating microglial polarization.

doi: 10.1016/j.intimp.2024.113786

Figure Lengend Snippet: Fig. 3. Effects of minocycline on the hippocampal neurons and synaptic plasticity of TN rats. (A) Representative Nissl staining images of hippocampal neurons (CA1, CA3, and DG areas), Scale bar = 20 μm. (B) The number of Nissl bodies in CA1, CA3 and DG regions. (C-I) NeuN, BDNF, TrkB, SYP, and PSD-95 protein levels were measured using Western blot analysis. Data are presented as mean ± SD (n = 3). ***P < 0.001/**P < 0.01/*P < 0.05 vs Sham; ##P < 0.01/#P < 0.05 vs dION-CCI.

Article Snippet: The primary antibodies used were against Neuronal nuclear antigen (NeuN) (ab60176, 1:1000, Abcam), Brain-derived neurotrophic factor (BDNF) (ab108319, 1:1000, Abcam), Tyrosine receptor kinase B (TrkB) (4603, 1:1000, Cell Signaling Technology, Danvers, MA, USA), Synaptophysin (SYP) (17785–1-AP, 1:6000, Proteintech Group, China), Postsynaptic density protein-95 (PSD95) (3409, 1:1000, Cell Signaling Technology), Iba1 (ab178846, 1:1000, Abcam), Cluster of Differentiation 86 (CD86) (sc-28347, 1:700, SantaCruz), iNOS (sc-7271, 1:500, SantaCruz), Arg1 (ab96183, 1:1000, Abcam), Mannose receptor (CD206) (24595, 1:1000, Cell Signaling Technology), TNF-α (AF7014, 1:500, Affinity, USA), IL-1β (AF5103, 1:1000, Affinity), IL-4 (sc-53084, 1:700, SantaCruz), IL-10 (DF6894, 1:1000, Affinity), TLR4 (sc-293072, 1:1000, SantaCruz), Myeloid differentiation primary response 88 (MyD88) (AF5195, 1:1000, Affinity), Nuclear factor-kappa B (NF-κb) (ABM0017, 1:1000, Abbkine, China), and β-actin (AF7018, 1:5000, Affinity).

Techniques: Staining, Western Blot