Journal: International Journal of Molecular Sciences

Article Title: Inhibition of Colony-Stimulating Factor 1 Receptor by PLX3397 Prevents Amyloid Beta Pathology and Rescues Dopaminergic Signaling in Aging 5xFAD Mice

doi: 10.3390/ijms21155553

Figure Lengend Snippet: Schematic diagram illustrating the experimental groups and protocol. When the 5xFAD mice were 9 months old, they were administered PLX3397 (50 mg·kg −1 , p.o. ) or vehicle for 30 days. Then, each group underwent positron emission tomography (PET) imaging targeting the dopamine D2 receptor (D2R) and metabotropic glutamate receptor 5 (mGluR5), and histopathological and molecular analyses were also performed.

Article Snippet: The washed tissues were then incubated with the following primary antibodies overnight at 4 °C: Anti-rabbit D2R (cat no. AB5084P; 1:500 dilution; Merck, Darmstadt, Germany), anti-rabbit TH (cat no. NB300-109; 1:1000 dilution; Novus, Centennial, CO, USA), or anti-rabbit DAT (cat no. D6944; 1:000 dilution; Sigma-Aldrich) antibodies.

Techniques: Positron Emission Tomography, Imaging

Journal: International Journal of Molecular Sciences

Article Title: Inhibition of Colony-Stimulating Factor 1 Receptor by PLX3397 Prevents Amyloid Beta Pathology and Rescues Dopaminergic Signaling in Aging 5xFAD Mice

doi: 10.3390/ijms21155553

Figure Lengend Snippet: Neuro-PET imaging for evaluating dopamine D2 receptor (D2R) and metabotropic glutamate receptor 5 (mGluR5) expression following PLX3397 treatment. ( A ) Representative nondisplaceable binding potential (BPND) parametric images for WT and Tg mice. Regional brain BPND levels in Tg mice after PLX3397 administration are shown. ( B ) Quantification of BPND for mGluR5 and D2R in the WT, Tg, and Tg + PLX3397 groups. * p < 0.05 WT vs. Tg; # p < 0.05 Tg vs. Tg + PLX3397 group. Error bars indicate SD ( n = 4/group).

Article Snippet: The washed tissues were then incubated with the following primary antibodies overnight at 4 °C: Anti-rabbit D2R (cat no. AB5084P; 1:500 dilution; Merck, Darmstadt, Germany), anti-rabbit TH (cat no. NB300-109; 1:1000 dilution; Novus, Centennial, CO, USA), or anti-rabbit DAT (cat no. D6944; 1:000 dilution; Sigma-Aldrich) antibodies.

Techniques: Imaging, Expressing, Binding Assay

Journal: International Journal of Molecular Sciences

Article Title: Inhibition of Colony-Stimulating Factor 1 Receptor by PLX3397 Prevents Amyloid Beta Pathology and Rescues Dopaminergic Signaling in Aging 5xFAD Mice

doi: 10.3390/ijms21155553

Figure Lengend Snippet: Effects of PLX3397 on the dopaminergic pathway. Representative immunohistochemical images for WT, Tg, and Tg + PLX3397 mice showing staining for dopamine D2 receptor (D2R) ( A ), tyrosine hydroxylase (TH) ( B ), and dopamine transporter (DAT) ( C ) and the quantification of D2R, TH, and DAT levels in the striatum region of the mouse brain ( n = 4 for TG group, n = 5 for WT and Tg + PLX3397 group). * p < 0.05 WT vs. Tg; # p < 0.05, ## p < 0.01 Tg vs. Tg + PLX3397 group. Error bars indicate SD.

Article Snippet: The washed tissues were then incubated with the following primary antibodies overnight at 4 °C: Anti-rabbit D2R (cat no. AB5084P; 1:500 dilution; Merck, Darmstadt, Germany), anti-rabbit TH (cat no. NB300-109; 1:1000 dilution; Novus, Centennial, CO, USA), or anti-rabbit DAT (cat no. D6944; 1:000 dilution; Sigma-Aldrich) antibodies.

Techniques: Immunohistochemical staining, Staining

Journal: Frontiers in Cellular Neuroscience

Article Title: Inhibitory Effects of Trifluoperazine on Peripheral Proinflammatory Cytokine Expression and Hypothalamic Microglia Activation in Obese Mice Induced by Chronic Feeding With High-Fat-Diet

doi: 10.3389/fncel.2021.752771

Figure Lengend Snippet: List of reagents (antibodies and assay kits) in the study.

Article Snippet: Polyclonal rabbit anti-D2R , Biorbyt, Cat# orb10515, RRID:AB_10747533.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Frontiers in molecular neuroscience

Article Title: Repeated inhibition of sigma-1 receptor suppresses GABA A receptor expression and long-term depression in the nucleus accumbens leading to depressive-like behaviors.

doi: 10.3389/fnmol.2022.959224

Figure Lengend Snippet: FIGURE 1 Repeated injection of NE-100 in the NAc induces depressive-like behaviors. (A) The time course of the experimental procedure. (B) Schematic diagram of the NAc (left panel) and image of the NAc injected with Evans-blue (right panel). (C) The level of σ1R mRNA in the NAc was examined at the end of the behavioral tests (n = 8 mice per group; Student’s t-test). (D) Representative Western blots of biotin-labeled surface proteins (surface) and total proteins (total) σ1R in the NAc. Na+-K+ ATPase served as internal control and GAPDH served as a negative control. Bar graphs indicate the ratio of proteins at the cell surface to their total levels. **p < 0.01 vs. control mice (n = 8 mice per group; Student’s t-test). (E) Bar graphs show the distance traveled in OFT in control mice and NE-100 mice (n = 12 mice per group; Student’s t-test). (F,G) Bar graphs show the immobility time of FST and TST in control mice and NE-100 mice. **p < 0.01 vs. control mice (n = 11 mice in control group of TST and n = 12 mice in other groups; Student’s t-test).

Article Snippet: Several antibodies were used in this study: anti-D2R (1:1,000; Millipore Cat#AB5084p, RRID:AB_2094980), anti-GABAAR-α1 (Sigma Cat#G4416, RRID:AB_477016), anti-GABAARα2(AbcamCat#ab72445, RRID:AB_1268929), anti-GABAAR-β2 (Affinity Biosciences Cat#DF6671, RRID:AB_2838633), antiGABAAR-β3 (Abcam Cat#ab98968, RRID:AB_10670809), anti-phosphorylation PKC (Epsilon Ser729; Abcam Cat#ab63387, RRID:AB_1142277), anti-PKC (Abcam Cat#ab253274, RRID:AB_2827663), anti-σ1R (B5; Santa Cruz Biotechnology Cat#sc-137075, RRID:AB_2285870), anti-Na+-K+ ATPase (Cell Signaling Technology Cat#3010, RRID:AB_2060983) and anti-GAPDH (Abcam Cat#ab181602, RRID:AB_2630358).

Techniques: Injection, Western Blot, Labeling, Control, Negative Control

Journal: Frontiers in molecular neuroscience

Article Title: Repeated inhibition of sigma-1 receptor suppresses GABA A receptor expression and long-term depression in the nucleus accumbens leading to depressive-like behaviors.

doi: 10.3389/fnmol.2022.959224

Figure Lengend Snippet: FIGURE 3 Repeated injection of NE-100 suppresses GABAAR and D2R expression. (A) The levels of GABAAR-α1-5, -β1-3, -γ1-3, -δ subunits, and D1R, D2R mRNA in the NAc were examined at the end of the behavioral tests. *p < 0.05 vs. control mice (n = 8 mice per group; two-way ANOVA). (B) Bar graphs show levels of GABAAR-α1, GABAAR-α2, GABAAR-β2, and GABAAR-β3 protein in the NAc normalized by the level of GAPDH were normalized by control levels. *p < 0.05 vs. control mice (n = 8 mice per group; two-way ANOVA). (C) Representative Western blots of biotin- labeled surface proteins (surface) and total proteins (total) D2R in the NAc. Na+-K+ ATPase served as internal control and GAPDH served as a negative control. Bar graphs indicate the ratio of proteins at the cell surface to their total levels. **p < 0.01 vs. control mice (n = 8 mice per group; Student’s t -test).

Article Snippet: Several antibodies were used in this study: anti-D2R (1:1,000; Millipore Cat#AB5084p, RRID:AB_2094980), anti-GABAAR-α1 (Sigma Cat#G4416, RRID:AB_477016), anti-GABAARα2(AbcamCat#ab72445, RRID:AB_1268929), anti-GABAAR-β2 (Affinity Biosciences Cat#DF6671, RRID:AB_2838633), antiGABAAR-β3 (Abcam Cat#ab98968, RRID:AB_10670809), anti-phosphorylation PKC (Epsilon Ser729; Abcam Cat#ab63387, RRID:AB_1142277), anti-PKC (Abcam Cat#ab253274, RRID:AB_2827663), anti-σ1R (B5; Santa Cruz Biotechnology Cat#sc-137075, RRID:AB_2285870), anti-Na+-K+ ATPase (Cell Signaling Technology Cat#3010, RRID:AB_2060983) and anti-GAPDH (Abcam Cat#ab181602, RRID:AB_2630358).

Techniques: Injection, Expressing, Control, Western Blot, Labeling, Negative Control

Journal: Frontiers in Cellular Neuroscience

Article Title: Inhibitory Effects of Trifluoperazine on Peripheral Proinflammatory Cytokine Expression and Hypothalamic Microglia Activation in Obese Mice Induced by Chronic Feeding With High-Fat-Diet

doi: 10.3389/fncel.2021.752771

Figure Lengend Snippet: List of reagents (antibodies and assay kits) in the study.

Article Snippet: Alternatively, brain sections were permeabilized in 0.3% Triton-X-100 in PBS for 30 min, incubated with rabbit anti-D2R antibody ( ) in PBS at 4°C overnight, and treated with biotinylated anti-rabbit IgG (1:200) in PBS for 1 h. The brain sections were then reacted with Vectastain ABC reagent (Vector Labs; Cat# PK-6100) containing 3,3’-diaminobenzidine and nuclear counterstaining by hematoxylin.

Techniques: Enzyme-linked Immunosorbent Assay