Journal: International journal of molecular sciences

Article Title: PM2.5 Exposure Triggers Hypothalamic Oxidative and ER Stress Leading to Depressive-like Behaviors in Rats.

doi: 10.3390/ijms252413527

Figure Lengend Snippet: Figure 1. Intranasal PM2.5 exposure induces ER stress, cell senescence, and reduced neuronal markers in specific rat brain regions. (A) Experimental timeline illustrating daily intranasal injections of either vehicle (0.9% NaCl) or PM2.5 (100 µg/20 µL) administered to Sprague–Dawley rats for 28 days, followed by behavioral tests. Vehicle, n = 5. PM2.5, n = 5. (B–E) qPCR analysis showing increased expression of ER stress and cell senescence markers (CHOP, eIF2α, GRP78, and P16) in the hypothalamus (B), hippocampus (C), cortex (D), and striatum (E) following PM2.5 exposure, with significant changes observed in the hypothalamus, cortex, and striatum. (F,G) PM2.5 exposure reduced mRNA expression of VAChT in the hypothalamus (F) but not in the hippocampus (G). (H) A representative immunoblot showing decreased VAChT protein level in the hypothalamus following PM2.5 treatment. β-actin served as a loading control. (I) Quantification of VAChT protein expression from panel H. Data are presented as mean ± SEM, with comparisons made using Student’s t-test in panels (B–G,I). ns., not significant. * p ≤0.05, ** p ≤0.01 vs. Vehicle, n = 5.

Article Snippet: The primary antibodies used for immunoblotting were: GAPDH (diluted 1:2500, sc-365062) and TH (diluted 1:1000, sc-25269) from Santa Cruz Biotechnology (Santa Cruz, CA, USA); VAChT (diluted 1:1000, #75-020) from NeuroMab Facility (UC Davis/NIH, CA, USA); β-actin (diluted 1:2500, ab6276) and β-III-tubulin (#ab7751, diluted 1:1000) from Abcam (Cambridge, UK); NOX4 (diluted 1:1000, NB110-58849) from Novus Biologicals (Littleton, CO, USA).

Techniques: Expressing, Western Blot, Control

Journal: International journal of molecular sciences

Article Title: PM2.5 Exposure Triggers Hypothalamic Oxidative and ER Stress Leading to Depressive-like Behaviors in Rats.

doi: 10.3390/ijms252413527

Figure Lengend Snippet: Figure 3. PM2.5 induces ER stress and mitochondrial ROS production, reducing neuronal markers in Neuro-2A cells. (A) q-PCR analysis of ER stress markers ATF4 and CHOP in Neuro-2A cells treated with low (1 µg/mL) and high (100 µg/mL) doses of PM2.5. (B) Western blot analysis of NOX4 protein levels in Neuro-2A cells treated with vehicle, 1 µg/mL, and 100 µg/mL of PM2.5, with β-actin as a loading control. (C,D) Mitochondrial ROS production in Neuro-2A cells treated with PM2.5 (100 µg/mL), showing a significant increase in ROS levels compared to vehicle control. (E,F) PM2.5- induced mitochondrial ROS increase was significantly reduced by NOX inhibitors GKT137831 and Apocynin. (G) Western blot analysis of β-III-tubulin and VAChT protein levels in Neuro-2A cells treated with increasing concentrations of PM2.5 (1, 3, 10, 30, and 100 µg/mL), with β-actin as a loading control. (H,I) Quantifying β-III-tubulin (H) and VAChT (I) protein levels from panel G, respectively. Scale bars, 5 µm. Bar graphs are expressed as mean ± SEM and analyzed using one-way ANOVA (A,D,F,H,I). * p ≤0.05, ** p ≤0.01, **** p ≤0.001 vs. Vehicle. # p ≤0.05 vs. PM2.5.

Article Snippet: The primary antibodies used for immunoblotting were: GAPDH (diluted 1:2500, sc-365062) and TH (diluted 1:1000, sc-25269) from Santa Cruz Biotechnology (Santa Cruz, CA, USA); VAChT (diluted 1:1000, #75-020) from NeuroMab Facility (UC Davis/NIH, CA, USA); β-actin (diluted 1:2500, ab6276) and β-III-tubulin (#ab7751, diluted 1:1000) from Abcam (Cambridge, UK); NOX4 (diluted 1:1000, NB110-58849) from Novus Biologicals (Littleton, CO, USA).

Techniques: Western Blot, Control

Journal: International Journal of Nanomedicine

Article Title: Bupivacaine Nanoparticles Inhibit Triple-Negative Breast Tumor Growth by Suppressing the Noradrenergic Nerves in Tumor Microenvironment

doi: 10.2147/IJN.S515895

Figure Lengend Snippet: Evaluation of neural type in orthotopic 4T1 tumor. ( A ) Representative immunofluorescence images of tumor tissues for the adrenergic nerve marker Tyrosine Hydroxylase (TH), cholinergic nerve marker vesicular acetylcholine transporter (VAChT) and sensory nerve marker calcitonin gene-related peptide (CGRP). Color coding is indicated at the top. Scale bars, 100 μm ( B ) the quantification of TH, CGRP and VAChT density of representative images. Data show mean ± SD; significant differences were calculated using One-way ANOVA followed by Tukey’s post-hoc test. **: P <0.01, *** P <0.001, ****: P <0.0001.

Article Snippet: Antigen retrieval was performed with citrate antigen retrieval solution (Beyotime, P0081) by heating slices in boiling temperature for 5 min. After cooling to room temperature, the slices were were blocked and permeabilized with PBS containing 10% goat serum and 0.3% Triton X-100 for 1 h. Then the slices were incubated with the following primary antibodies at 4°C overnight: F4/80 antibody (1:200, Abcam, ab6640); anti-TH antibody (1:600, Merck, AB152); anti-VAChT antibody (1:200, Novus Biologicals, NB110-74,764); anti-CGRP (1:200, Thermo Fisher Scientific, PA5-114929).

Techniques: Immunofluorescence, Marker

Journal: The Journal of Neuroscience

Article Title: Cholinergic Neurotransmission in the Posterior Insular Cortex Is Altered in Preclinical Models of Neuropathic Pain: Key Role of Muscarinic M2 Receptors in Donepezil-Induced Antinociception

doi: 10.1523/JNEUROSCI.1537-15.2015

Figure Lengend Snippet: Differentially expressed genes in the posterior insular cortex of oxaliplatin-treated rats compared with controls using TLDA a

Article Snippet: Rn00581454_s1 , Slc18a3 , Solute carrier family 18 (vesicular acetylcholine), member 3 (VAChT) , 0.38 , 0.401.

Techniques:

Journal: Neural Regeneration Research

Article Title: Small molecule inhibitor DDQ-treated hippocampal neuronal cells show improved neurite outgrowth and synaptic branching

doi: 10.4103/NRR.NRR-D-24-00157

Figure Lengend Snippet: Antibodies used in immunofluorescence analysis

Article Snippet: VAChT , Rabbit polyclonal 1:200 , ACT-003-200UL , Alomone Labs Jerusalem BioPark (JBP) Jerusalem, Israel , , Goat-anti-rabbit Alexa Fluor488 1:400 , A-11008 , Thermo Fisher Scientific.

Techniques: Immunofluorescence

Journal: Biochemical pharmacology

Article Title: CDP-choline modulates cholinergic signaling and gut microbiota to alleviate DSS-induced inflammatory bowel disease.

doi: 10.1016/j.bcp.2023.115845

Figure Lengend Snippet: Fig. 3. Effect of CDP-choline on ACh synthesis-related proteins in colonic tissue of mice. (A) The representative image of ChT1, ChAT, VAChT, AChE, TNF-α, and IL-6 expression among the three groups. Histogram of the relative expressions of ChT1 (B), ChAT (C), VAChT (D), AChE (E), TNF-α (F), and IL-6 (G), respectively (n = 6). (H) Histogram of ChAT activity in the colon among the three groups measured by ELISA (n = 8). Data are expressed as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The membranes were sealed (20 °C, 12 min) in protein-free rapid blocking buffer (EpiZyme, PS108P, China) and incubated overnight at 4 °C in the following primary antibody: β-Actin (1:1000, Cell Signaling Technology, #3700, USA), Occludin (1:1000, Cell Signaling Technology, #91131, USA), High-affinity choline transporter (SLC5A7/ChT1 1:1000, ThermoFisher, PA5-42485, USA), Choline acetyltransferase (ChAT, 1:1000, Cell Signaling Technology, #27269, USA), Acetylcholinesterase (AChE, 1:1500, Abcam, ab183591, UK), Vesicular acetylcholine transporter (SLC18A3/VAChT, 1:1000, Boster Bio, A04891, China), α7 nicotinic acetylcholine receptor (CHRNA7/α7 nAChR, 1:1000, Proteintech, 21379–1-AP, China), Tumor necrosis factor-α (TNF-α, 1:1000, Cell Signaling Technology, #11948, USA), Interleukin 6 (IL-6, 1:1000, HUABIO, R1412-2, China) and ZO-1 (1:1500, Bioswamp, PAB36669, China).

Techniques: Expressing, Activity Assay, Enzyme-linked Immunosorbent Assay

Journal: JARO: Journal of the Association for Research in Otolaryngology

Article Title: Transient Expression of Acetylcholinesterase in the Posterior Ventral Cochlear Nucleus of Rat Brain

doi: 10.1007/s10162-004-5015-4

Figure Lengend Snippet: Immunohistochemistry was conducted using an antibody to the vesicular acetylcholine transport protein (VAChT). A. A representative sample at P7, when AChE expression is at peak levels. VAChT was not visualized in the PVCN at any postnatal age investigated (P3–15). B. In the same tissue section, VAChT was readily visualized in many other cell groups, including the facial nucleus shown here.

Article Snippet: The primary antibody (goat anti-rat VAChT; ImmunoStar) was used in dilutions of 1/100 to 1/10,000.

Techniques: Immunohistochemistry, Expressing