Journal: Biomolecules & Therapeutics

Article Title: α-Pinene Attenuates Methamphetamine-Induced Conditioned Place Preference in C57BL/6 Mice

doi: 10.4062/biomolther.2022.132

Figure Lengend Snippet: Effect of APN on METH-decreased TH and D2R in striatum of C57BL/6 mice. Expression of TH (A) and D2R (B) in METH-treated mice was compared with saline- or APN-treated mice. The relative intensity of TH/Actin and D2R/Actin was represented in the right panel. Data were expressed as mean ± SEM. Significantly different between groups: ** p <0.01 saline-treated control mice vs METH-treated mice, # p <0.05 and ## p <0.01 METH-treated mice vs APN-treated mice (n=5).

Article Snippet: Anti-D2R antibody was supplied by Biorbyt LLC. (Wobum, MA, USA) and Bioss Antibodies Inc. (Woburn, MA, USA), respectively.

Techniques: Expressing, Saline, Control

Journal: Scientific Reports

Article Title: Gestational stress disrupts dopamine and oxytocin signaling in the postpartum reward system of rats: implications for mood, motivation and mothering

doi: 10.1038/s41598-024-84043-6

Figure Lengend Snippet: Experimental timeline. Pregnant rats were exposed to a chronic variable stress procedure from gestation day 7 (GD7)-GD20. No stress controls were handled daily. The day of birth (DOB) was designated as postpartum day 0 (PD0). Behavioral testing and brain collection occurred from PD2-PD10. In Expt. 1 , rats (No Stress = 10; Stress = 9) underwent testing for maternal behavior and brains were collected on PD7 for qPCR gene expression analysis of oxytocin (OT) and oxytocin receptor (OTR) in the hypothalamus and ventral tegmental area (VTA). In Expt. 2, rats (No Stress = 6; Stress = 5) were tested on the conditioned place preference (CPP) paradigm. In Expt. 3 , rats (No Stress = 9; Stress = 9) underwent testing on the sucrose preference test (SPT) and brains were collected on PD8 for immunohistochemistry (IHC) and densiometric analysis of dopamine (DA) markers (DAT, dopamine transporter; D1R; dopamine 1 receptor; D2R, dopamine 2 receptor; TH, tyrosine hydroxylase) in the NAc. Maternal and litter characteristics were measured in the SPT dams. In Expt. 4 , rats (No Stress = 12; Stress = 10) were tested on the elevated plus maze (EPM) and forced swim test (FST) and brains were collected on PD10 for IHC and densiometric analysis of additional DA markers in the NAc (pTH, phosphorylated tyrosine hydroxylase; VMAT, vesicular monoamine transporter) as well as counts of TH + cells in the VTA. OT IHC was also done to assess the effects of gestational stress on OT + cells in the hypothalamus and OT fibers in the VTA. In Expt. 5 , dams (No Stress = 6; Stress = 6), the NAc was dissected on PD6 for Liquid Chromatography-Mass Tandem Spectrometry (LC–MS/MS) to measure DA content and the dopamine metabolite, DOPAC. Figure created in https://BioRender.com .

Article Snippet: Next, sections were blocked in 5% BSA in 0.1 M PBS for 1 h at RT and incubated with one of the following primary antibodies: rabbit anti-D1R (1:50; Cat#sc-14001, Santa Cruz Biotechnology), goat anti-dopamine D2R (1:100; Cat#sc-7522, Santa Cruz Biotechnology), rabbit anti-TH (1:1000; Cat#AB152, Millipore), rabbit anti-DAT (1:200; Cat#AB1591P, Millipore), rabbit anti-VMAT(1:100; Cat#PA5-22,864, ThermoFisher), and rabbit anti- pTH (1:500; Cat#AB5935, Millipore) overnight at 4 °C.

Techniques: Gene Expression, Conditioned Place Preference, Immunohistochemistry, Liquid Chromatography, Liquid Chromatography with Mass Spectroscopy

Journal: Scientific Reports

Article Title: Gestational stress disrupts dopamine and oxytocin signaling in the postpartum reward system of rats: implications for mood, motivation and mothering

doi: 10.1038/s41598-024-84043-6

Figure Lengend Snippet: Gestational stress alters markers of dopaminergic signaling in the NAc shell. ( A ) In the nucleus accumbens (NAc) shell, markers associated with dopaminergic signaling were examined via immunohistochemistry on PD8 or PD10. Compared to unstressed controls, dams exposed to gestational stress showed a reduction in percent area staining for several dopaminergic markers including ( B ) tyrosine hydroxylase, TH, ( C ) phosphorylated TH, pTH and ( D ) the dopamine transporter, DAT. ( E ) There was no effect of gestational stress on vesicular monoamine transporter, VMAT. Percent area staining for dopamine receptors ( F ) D1R and ( G ) D2R in the NAc shell was also examined and only the D2R was reduced by gestational stress. Representative images of each marker analyzed are shown in ( A ). * p < 0.05; ** p < 0.005; *** p ≤ 0.0005.

Article Snippet: Next, sections were blocked in 5% BSA in 0.1 M PBS for 1 h at RT and incubated with one of the following primary antibodies: rabbit anti-D1R (1:50; Cat#sc-14001, Santa Cruz Biotechnology), goat anti-dopamine D2R (1:100; Cat#sc-7522, Santa Cruz Biotechnology), rabbit anti-TH (1:1000; Cat#AB152, Millipore), rabbit anti-DAT (1:200; Cat#AB1591P, Millipore), rabbit anti-VMAT(1:100; Cat#PA5-22,864, ThermoFisher), and rabbit anti- pTH (1:500; Cat#AB5935, Millipore) overnight at 4 °C.

Techniques: Immunohistochemistry, Staining, Marker

Journal: International Journal of Molecular Sciences

Article Title: Alteration of the Dopamine Receptors’ Expression in the Cerebellum of the Lysosomal Acid Phosphatase 2 Mutant (Naked–Ataxia ( NAX )) Mouse

doi: 10.3390/ijms21082914

Figure Lengend Snippet: RNA sequencing data analysis in nax ( n = 5) and wild–type (wt) ( n = 6) mice. Data analysis shows dopamine receptor D1 ( Drd1, A ), Drd3 ( C ), and Drd4 ( D ) are significantly increased in the nax cerebellum. Although an increase in Drd2 ( B ) and Drd5 ( E ) is apparent, statistical significance was not reached. The data in the bar graph are presented as the mean ± SEM, and statistical analysis was performed using an unpaired t –test (* p < 0.05 and ** p < 0.01). RPKM: Reads Per Kilobase of transcript per Million mapped reads.

Article Snippet: D1: rabbit polyclonal anti–D1 dopamine receptor (TA328798, anti–Drd1, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA); D2: rabbit polyclonal anti–D2 dopamine receptor (TA328800, anti–Drd2, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA), produced against recombinant rat dopamine receptor 2 (DR2); D3: rabbit polyclonal anti–D3 dopamine receptor (TA328800, anti–Drd3, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA), produced against recombinant rat dopamine receptor 3 (DR3); D4: rabbit polyclonal anti–D4 dopamine receptor (TA321202, anti–DRD4, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA), produced against recombinant rat dopamine receptor D4 (DRD4); and D5: rabbit polyclonal anti–D5 dopamine receptor (TA328802, anti–Drd5, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA), produced against recombinant rat dopamine receptor 5 (DR5).

Techniques: RNA Sequencing Assay

Journal: International Journal of Molecular Sciences

Article Title: Alteration of the Dopamine Receptors’ Expression in the Cerebellum of the Lysosomal Acid Phosphatase 2 Mutant (Naked–Ataxia ( NAX )) Mouse

doi: 10.3390/ijms21082914

Figure Lengend Snippet: Frontal sections of wt and nax mouse cerebella: DRD2 expression at P5 and P17. Immunoperoxidase staining for DRD2 in wt ( A ) and nax ( B ) cerebella at P5 shows weak immunoreactivity in the Pcl of wt but relatively strong immunoreactivity in PC somata of the nax cerebellum. ( C , D ) Immunoperoxidase staining for DRD2 in wt ( C ) and nax ( D ) cerebella at P17 shows weak immunoreactivity in PCs, with no differences between the two groups. ( E ) Western blot analysis of whole cerebellar lysates revealed no significant differences in DRD2 protein expression between wt and nax cerebella at P5 and P17 (wt: n = 3 and nax : n = 3). The data in the bar graphs are presented as the means of three independent experiments ± SEM; statistical analysis was performed using two–way ANOVA. P; postnatal. Scale bars: 100 μm in A, C, and D and 50 μm in B.

Article Snippet: D1: rabbit polyclonal anti–D1 dopamine receptor (TA328798, anti–Drd1, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA); D2: rabbit polyclonal anti–D2 dopamine receptor (TA328800, anti–Drd2, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA), produced against recombinant rat dopamine receptor 2 (DR2); D3: rabbit polyclonal anti–D3 dopamine receptor (TA328800, anti–Drd3, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA), produced against recombinant rat dopamine receptor 3 (DR3); D4: rabbit polyclonal anti–D4 dopamine receptor (TA321202, anti–DRD4, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA), produced against recombinant rat dopamine receptor D4 (DRD4); and D5: rabbit polyclonal anti–D5 dopamine receptor (TA328802, anti–Drd5, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA), produced against recombinant rat dopamine receptor 5 (DR5).

Techniques: Expressing, Immunoperoxidase Staining, Western Blot