Journal: Ecotoxicology and environmental safety

Article Title: Chronic developmental exposure to low-dose ([C8mim][PF6]) induces neurotoxicity and behavioural abnormalities in rats.

doi: 10.1016/j.ecoenv.2021.112806

Figure Lengend Snippet: Fig. 4. Developmental exposure to low-dose ([C8MIM] [PF6]) increased the expression of NMDA receptors. (A) The mRNA level of major receptors (nmdar1, nmdar2a, nmdar2b for NMDA receptor; drd1 and drd2 for dopamine receptor; 5-ht2ar for 5-HT receptor; gabaarα1 for GABA receptor) for main neurotransmitters in hippocampus and prefrontal cortex. (B) The protein levels of NMDA receptors (NMDAR1, NMDAR2A, and NMDAR2B) in the hippocam pus and prefrontal cortex. (C) The protein levels of dopa mine receptors (DRD1 and DRD2), 5-HT receptors (5- HT2AR), and GABA receptors (GABAARα1) in the hippo campus and prefrontal cortex. (D) Statistical analysis of protein expression. n = 3 for each group. Student’s t-test was performed for the analysis. The data are presented as the mean ± SEM. *p＜0.05, **p＜0.01, ***p＜0.001.

Article Snippet: Membranes were treated with 5% skim milk for 1 h, followed by incubation with primary antibodies against NMDAR1 (ab109182, Abcam), NMDAR2A (ab124913, Abcam), NMDAR2B (ab65783, Abcam), DRD1 (ab81296, Abcam), DRD2 (ab30743, Abcam), 5-HT2AR (PB9599, Boster), GABAARα1 (ab252430, Abcam), PSD95 (36233S, Cell Signalling Technology), IBA1 (GTX100042, GeneTex), GFAP (BA0056, Boster), NeuN (ab177487, Abcam), MBP (BA0094, Boster), IFN-γ (PAA033Ra01, Cloud-clone Corp.), IL-1β (PAA563Ra01, Cloud-clone Corp.), TNF-α (PAA133Ra01, cloud-clone corp.), NF-ĸB (66535-1-lg, Proteintech), Histone H3 (AF0009, Beyotime), NRF2 (bs-1074R-1, bioss), PPARγ(ab272718, Abcam), COX2 (ab179800, Abcam), iNOS (ab178945, Abcam), Caspase 3 (9662s, CST), Bax (SC-7480, SANTA CRUZ), Bcl-2 (sc7382, SANTA CRUZ), GAPDH (5174T, CST), and β-actin (3700s, CST) at a 1:1000 dilution overnight at 4 ◦C.

Techniques: Expressing

Journal: Frontiers in Psychiatry

Article Title: Histone deacetylase inhibitors mitigate antipsychotic risperidone-induced motor side effects in aged mice and in a mouse model of Alzheimer’s disease

doi: 10.3389/fpsyt.2022.1020831

Figure Lengend Snippet: Effects of histone deacetylase (HDAC) inhibitors on histone acetylation at the 5htr2a and Drd2 gene promoters in the striatum in young and aged mice. (A) No significant differences of H3K27ac binding at the 5htr2a promoter between young and aged group was found, but a significant increase in H3K27ac binding at the 5ht2a promoter in the striatum was found in aged and young mice co-treated with RIS and VPA or MS-275 compared to those treated with RIS alone. (B) H3K27ac binding at the Drd2 promoter was significantly decreased in aged mice as compared to young mice in Veh treated group. However, H3K27ac binding at the Drd2 promoter in the striatum was observed to be significantly increased in aged mice co-treated with RIS and HDAC inhibitors VPA or MS-275 when compared to those treated with Veh alone. Data are presented as mean ± SEM, ( n = 5). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Blots were blocked for 1-hour with 5% non-fat dry milk (Biorad, 1706404) and incubated using 1:1,000 5HT2A (Boster Biological, PA1373), 1:500 D2R (Millipore, AB5084P), or 1:1000 B-actin (Santa Cruz, sc-47778) overnight at 4°C.

Techniques: Histone Deacetylase Assay, Binding Assay

Journal: Pharmacology, biochemistry, and behavior

Article Title: Chronic methamphetamine self-administration dysregulates 5-HT2A and mGlu2 receptor expression in the rat prefrontal and perirhinal cortex: comparison to chronic phencyclidine and MK-801.

doi: 10.1016/j.pbb.2018.09.007

Figure Lengend Snippet: Identification of antibodies suitable for the detection of 5-HT2A and mGlu2 receptor proteins in the rat brain tissue. (A) Validation of six different commercially-available antibodies in the frontal cortex of rats (R), wild-type mice (WT) and 5-HT2A knockout mice (KO). Only one antibody (Immunostar #24288) detected a ~75kDa band (◀) corresponding to a mature, glycosylated rat 5-HT2A receptor that was not present in 5-HT2A KO mice. (B) Deglycosylation of the wild type (WT) mouse cortical lysate (lanes 2 and 3) by PNGase F increased electrophoretic mobility of the ~75 kDa (◀) band to yield ~52 kDa (◁) band (detected by Immunostar #24288 5-HT2A receptor antibody) that corresponds to the sequenced-based molecular weight of the rat 5-HT2A receptor. For a comparison, the 5-HT2A receptor detection (Immunostar #24288) in the rat (R) cerebral cortex has been shown (lane 1). (C) Immunostaining of rat cortical sections with 5-HT2A #24288 antibody produced a laminar staining pattern consistent with the known 5-HT2A distribution in the mPFC (as shown under 10× magnification). This signal was not present in the mPFC slices, or in the mPFC tissue homogenates incubated with the 5-HT2A antiserum preadsorbed with its immunogenic peptide.

Article Snippet: Only one antibody (Immunostar #24288) detected a ~75kDa band (◀) corresponding to a mature, glycosylated rat 5-HT2A receptor that was not present in 5-HT2A KO mice. (B) Deglycosylation of the wild type (WT) mouse cortical lysate (lanes 2 and 3) by PNGase F increased electrophoretic mobility of the ~75 kDa (◀) band to yield ~52 kDa (◁) band (detected by Immunostar #24288 5-HT2A receptor antibody) that corresponds to the sequenced-based molecular weight of the rat 5-HT2A receptor.

Techniques: Biomarker Discovery, Knock-Out, Molecular Weight, Comparison, Immunostaining, Produced, Staining, Incubation