mouse monoclonal anti d2r  (Santa Cruz Biotechnology)

Journal: Frontiers in Pharmacology

Article Title: Evidence for the existence of facilitatory interactions between the dopamine D2 receptor and the oxytocin receptor in the amygdala of the rat. Relevance for anxiolytic actions

doi: 10.3389/fphar.2023.1251922

Figure Lengend Snippet: Detection of D2R–OXTR heteroreceptor complexes in the different subfields of rat amygdala by in situ proximity ligation assay ( in situ PLA). The nuclei are shown in blue (DAPI) and the positive PLA puncta/blobs (D2R–OXTR heteroreceptor complexes) in red. (Left panel) Quantitative analysis of the D2R–OXTR heteroreceptor complex expression in the CeA and BLA of the rat brain detected by means of the in situ PLA. PLA was quantified as PLA (red puncta/blobs) per nuclei per field by an experimenter blind to the experiment conditions. The analysis (means ± S.E.M.) was performed on images obtained from at least two slices per animal (four rats in total). Statistical analysis was performed by Student’s t-test with Bonferroni correction. The p -value of 0.05 and lower was considered significant: * p < 0.05. Bregma 1.00 mm, Scale bar is 30 μm. (Right panels) Representative confocal image of the in situ PLA at the level of the CeA and BLA. Some positive PLA clusters/blobs are indicated by an arrow, and the scale bar for this panel is indicated on the right bottom of the panel.

Article Snippet: The primary antibodies used were goat polyclonal anti-oxytocin receptor (5 μg/mL; ab87312 from Abcam, Sweden) and mouse monoclonal anti-D2R (MABN53, 1:600, Millipore, Sweden).

Techniques: In Situ, Proximity Ligation Assay, Expressing

Journal: Frontiers in Pharmacology

Article Title: Evidence for the existence of facilitatory interactions between the dopamine D2 receptor and the oxytocin receptor in the amygdala of the rat. Relevance for anxiolytic actions

doi: 10.3389/fphar.2023.1251922

Figure Lengend Snippet: BRET2 analysis was conducted to examine the heterodimerization of D2R and OXTR receptors. (A) Transfection of HEK293T cells with a constant DNA concentration of acceptor D2R-Rluc and increasing concentrations of donor OXTR–GFP2 constructs. BRET2 ratio, total fluorescence, and total luminescence were determined as described in the Materials and Methods section. As a negative control, cells individually expressing D2R-Rluc were mixed with cells individually expressing OXTR-GFP2 prior to exposure to coelenterazine-400a. The X -axis represents the fluorescence value obtained from GFP2, normalized with the luminescence value of D2R-Rluc expression 10 min after coelenterazine incubation. Ten saturation curves were plotted, and a nonlinear regression equation assuming a single binding site was used to fit the curves. The D2R–OXTR curve exhibited a better fit to a saturation curve compared to a linear regression, as demonstrated by the F-test ( p < 0.01). This finding contrasts with the mixed pool of cells expressing D2R-Rluc and OXTR-GFP2. The data presented are the mean ± S.E.M., with n = 15–20, performed in triplicate. (B) Evaluation of the effects of 20 min of stimulation with different D2R and OXTR agonists and antagonists, as well as combined treatments, on the BRET ratios for D2R–OXTR heteroreceptor complexes. The ratios are expressed as the mean ± S.E.M. from at least six experiments. Combination treatment with quinpirole and oxytocin shows statistically significant difference compared with the vehicle group, one-way ANOVA, Bonferroni post-test.

Article Snippet: The primary antibodies used were goat polyclonal anti-oxytocin receptor (5 μg/mL; ab87312 from Abcam, Sweden) and mouse monoclonal anti-D2R (MABN53, 1:600, Millipore, Sweden).

Techniques: Transfection, Concentration Assay, Construct, Fluorescence, Negative Control, Expressing, Incubation, Binding Assay

Journal: PLoS ONE

Article Title: Iron-Responsive Olfactory Uptake of Manganese Improves Motor Function Deficits Associated with Iron Deficiency

doi: 10.1371/journal.pone.0033533

Figure Lengend Snippet: Rats intranasally instilled with MnCl 2 (6×10 mg/kg) were euthanized and striatal tissues were collected and homogenized to determine the expression levels of dopamine transporter (DAT; A ), dopamine receptor D 1 (D1R; B ), and dopamine receptor D 2 (D2R; C ). Relative intensities of protein bands normalized to actin were determined using Odyssey software (version 2.1). Empty and closed bars represent water-instilled and MnCl 2 -instilled rats, respectively. Data were presented as mean ± SEM (N = 3–4 per group) and were analyzed using two-way ANOVA.

Article Snippet: After blocking, the membrane was incubated in goat anti-dopamine transporter (DAT) antibody (1∶100, Santa Cruz), mouse anti-dopamine receptor D 2 (D2R) antibody (1∶100, Santa Cruz) or rat anti-dopamine receptor D 1 (D1R) antibody (1∶100, Sigma).

Techniques: Expressing, Software