Journal: Frontiers in Neural Circuits

Article Title: Differential Expression of Dopamine D5 Receptors across Neuronal Subtypes in Macaque Frontal Eye Field

doi: 10.3389/fncir.2018.00012

Figure Lengend Snippet: Information on primary antibodies used for immunofluorescence.

Article Snippet: We performed two controls to verify the specifity of the D5R antibody from Alomone Labs (ADR-005, see Supplementary Figure ).

Techniques: Immunofluorescence

Journal: Frontiers in Neural Circuits

Article Title: Differential Expression of Dopamine D5 Receptors across Neuronal Subtypes in Macaque Frontal Eye Field

doi: 10.3389/fncir.2018.00012

Figure Lengend Snippet: Distribution of D5Rs across cortical layers. (A) Number of NeuN+ neurons that co-express D5R for cortical layers I, II–III, IV, V and VI per mm 2 . (B) Proportion of NeuN+ neurons that express D5Rs for cortical layers I, II–III, IV, V and VI.

Article Snippet: We performed two controls to verify the specifity of the D5R antibody from Alomone Labs (ADR-005, see Supplementary Figure ).

Techniques:

Journal: Frontiers in Neural Circuits

Article Title: Differential Expression of Dopamine D5 Receptors across Neuronal Subtypes in Macaque Frontal Eye Field

doi: 10.3389/fncir.2018.00012

Figure Lengend Snippet: D5R expression on different cell types. (A) Co-expression of D5Rs (green) and SMI-32+ putative long-range projection pyramidal neurons (red; left) and co-expression of D5Rs with a general pyramidal neuron stain (neurogranin, red; right). (B) Proportion of frontal eye field (FEF) SMI-32+ and Neurogranin+ neurons that express D5Rs. (C) Expression of D5Rs (green) among different interneuron subtypes (red): parvalbumin+ (top left), calretinin+ (top right), calbindin+ (bottom left), somatostatin+ (bottom right). (D) Proportion of inhibitory interneurons that express D5Rs. Statistical comparisons were made between parvalbumin+, calbindin+ and calretinin+ neurons. Somatostatin+ neurons were excluded because they were too sparse to be used in statistical comparisons. For all panels: scale bar is equal to 100 μm and *** denotes significance at the level p ≤ 0.001.

Article Snippet: We performed two controls to verify the specifity of the D5R antibody from Alomone Labs (ADR-005, see Supplementary Figure ).

Techniques: Expressing, Staining

Journal: Frontiers in Neural Circuits

Article Title: Differential Expression of Dopamine D5 Receptors across Neuronal Subtypes in Macaque Frontal Eye Field

doi: 10.3389/fncir.2018.00012

Figure Lengend Snippet: Proportion of D5Rs on different cell types across cortical layers. (A) Neurogranin+ pyramidal neurons and SMI-32+ putative long-range projection neurons. (B) Parvalbumin+, calbindin+, calretinin+ and somatostatin+ inhibitory interneurons. Proportions are broken down by cortical layer (I, II–III, IV, V and VI) and compared to SMI-32+ D5R+ proportions (light gray bars). *, ** and *** indicate significance at the levels of p ≤ 0.05, 0.01 and 0.001 (Bonferonni-adjusted values), respectively. Differences in the proportion of D5R+ neurons across layers were also calculated individually for each cell type. Only calretinin+ neurons exhibited a significant difference in D5R+ proportions across layers: indicated with a vertical line spanning layers II-VI and significance at the level of p ≤ 0.001 (Bonferonni-adjusted value) is indicated by ††† .

Article Snippet: We performed two controls to verify the specifity of the D5R antibody from Alomone Labs (ADR-005, see Supplementary Figure ).

Techniques:

Journal: Frontiers in Neural Circuits

Article Title: Differential Expression of Dopamine D5 Receptors across Neuronal Subtypes in Macaque Frontal Eye Field

doi: 10.3389/fncir.2018.00012

Figure Lengend Snippet: Proportion of D5R+ neurons by cell type for different cortical layers. For cortical layers I, II–III, IV, V and VI: proportion of D5R+ cells that express a given cell type marker (SMI-32, neurogranin, parvalbumin, calretinin, somatostatin).

Article Snippet: We performed two controls to verify the specifity of the D5R antibody from Alomone Labs (ADR-005, see Supplementary Figure ).

Techniques: Marker

Journal: Frontiers in Pharmacology

Article Title: Dopamine internalization via Uptake 2 and stimulation of intracellular D 5 -receptor-dependent calcium mobilization and CDP-diacylglycerol signaling

doi: 10.3389/fphar.2024.1422998

Figure Lengend Snippet: Effects of dopamine (DA) microinjection or bath application on intracellular free calcium concentrations [iCa 2+ ] in hDRD5 receptor-expressing cells. (A) . Human U2-OS osteosarcoma cells natively expressing DRD1 and PMAT, but not DRD5, were pre-transfected with GFP-fluorescent human DRD5 receptor (green GFP-D5 fluorescence). Dopamine and other treatments were microinjected into the cell Soma followed by continuous 0.25 Hz recording of [iCa 2+ ] signal as indicated by Fura-2AM. fluorescence. Pictograms of [iCa 2+ ] from respective representative cells are shown (at least 12 transfected cells were tested with similar outcomes, and average results from 12 cells are shown in part D below). The control cell tested in parallel (top panel) was microinjected with buffered media alone, while the DA injected cell received 10 nM final intracellular concentration of DA. The [iCa 2+ ] signal integrated over a 6 min observation window as well as point observations at 2 and 6 min, are shown. (B) . Effects of co-microinjection of SCH23390 (SCH) 10 nM with dopamine 10 nM (Top panel) or of quinpirole microinjection (up to 10 μM, bottom panel) on [iCa 2+ ] response. SCH23390 blocked the iCa 2+ response to dopamine, while the D 2 agonist quinpirole (Quin) was without effect. (C) . Pictograms from representative experiments testing the effects of bath-applied DA on iCa 2+ mobilization in U2-OS cells. Bath application of 10 nM dopamine had no effect (Top panel), while 10 µM dopamine monitored over 11 min revealed a slowly rising wave of [iCa 2+ ] response that was highest toward the latest 11 min time point observed (Bottom panel). (D) . Quantified observations and temporal patterns of responses for microinjected versus bath applied dopamine. Continually recorded [iCa 2+ ] data resampled at 0.133 min intervals from 12 experiments are shown for microinjected DA 10 nM (Left graph, peak effect = 416%) in contrast with the effects of bath-applied DA 10 nM (Middle graph) and bath-applied DA 10 µM (Right graph). For bath-applied DA 10 μM, the average of the three observations made in the last 12 s of recording (106.2%, N = 12) was compared to the average of the three observations made in the first 12 s (0.2 min) of recording as the baseline (100%, N = 12); the comparison indicated a statistically significant increase in bath-applied dopamine-stimulated [iCa 2+ ] response (paired t -test, p = 0.019). The experimental system could not support extension of observation period past the 11th minute. (E) Expression indications for various dopamine receptors and PMAT in U2-OS cells. While D 1 and D 2 receptors as well as PMAT were expressed, there was no indication of D 5 receptor expression in the U2-OS cells.

Article Snippet: U2-OS cells were transfected with GFP-tagged human D 5 receptor (DRD5) cDNA ( OriGene.com , #RG202502) or pCMV6-AC-mGFP vector ( OriGene.com #PS100040) using Turbofectin-8 transfection reagent ( Origene.com #TF81001) according to manufacturer recommendations.

Techniques: Microinjection, Expressing, Transfection, Fluorescence, Control, Injection, Concentration Assay, Comparison

Journal: Frontiers in Neural Circuits

Article Title: Differential Expression of Dopamine D5 Receptors across Neuronal Subtypes in Macaque Frontal Eye Field

doi: 10.3389/fncir.2018.00012

Figure Lengend Snippet: Information on primary antibodies used for immunofluorescence.

Article Snippet: D5R , Dopamine D5 receptor , 1:200 , Rabbit Polyclonal; ADR-005; Alomone Labs.

Techniques: Immunofluorescence