anti d5 dopamine receptor extracellular antibody  (Alomone Labs)


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    Alomone Labs anti d5 dopamine receptor extracellular antibody
    The deficiency of <t>DRD5</t> signaling increased colonic M1 macrophages but reduced M2 cells in the colon of DSS colitis mice. A Immunofluorescent labeling of F4/80 + , Arg1 + , and Inos + colonic macrophages of WT and DRD5 −/− mice on day 6 after DSS treatment (top). The cell numbers are quantified by Image pro plus (bottom) ( n = 3). Scale bars, 200 μm. B Flow cytometry analysis of colonic macrophages of WT and DRD5 −/− mice ( n = 5 mice per group) on day 6 after DSS treatment as indicated. Data are presented as representative plots (top) and summary graphs of quantified percentages (bottom). C Flow cytometry analysis of the CD45.2 + cells percentage in Arg1 + and CD86 + colonic macrophages of lethally irradiated mice reconstituted with DRD5 −/− CD45.2/WT CD45.1 bone marrow (1:1 ratio), and a control WT CD45.2/WT CD45.1 group (1:1 ratio) on day 6 after DSS treatment ( n = 3 mice per group). Data are presented as representative plots (left) and summary graphs of quantified percentages (right). Data are pooled from two or three independent experiments. Error bars show means ± SEM. * p
    Anti D5 Dopamine Receptor Extracellular Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti d5 dopamine receptor extracellular antibody - by Bioz Stars, 2022-12
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    Images

    1) Product Images from "DA-DRD5 signaling controls colitis by regulating colonic M1/M2 macrophage polarization"

    Article Title: DA-DRD5 signaling controls colitis by regulating colonic M1/M2 macrophage polarization

    Journal: Cell Death & Disease

    doi: 10.1038/s41419-021-03778-6

    The deficiency of DRD5 signaling increased colonic M1 macrophages but reduced M2 cells in the colon of DSS colitis mice. A Immunofluorescent labeling of F4/80 + , Arg1 + , and Inos + colonic macrophages of WT and DRD5 −/− mice on day 6 after DSS treatment (top). The cell numbers are quantified by Image pro plus (bottom) ( n = 3). Scale bars, 200 μm. B Flow cytometry analysis of colonic macrophages of WT and DRD5 −/− mice ( n = 5 mice per group) on day 6 after DSS treatment as indicated. Data are presented as representative plots (top) and summary graphs of quantified percentages (bottom). C Flow cytometry analysis of the CD45.2 + cells percentage in Arg1 + and CD86 + colonic macrophages of lethally irradiated mice reconstituted with DRD5 −/− CD45.2/WT CD45.1 bone marrow (1:1 ratio), and a control WT CD45.2/WT CD45.1 group (1:1 ratio) on day 6 after DSS treatment ( n = 3 mice per group). Data are presented as representative plots (left) and summary graphs of quantified percentages (right). Data are pooled from two or three independent experiments. Error bars show means ± SEM. * p
    Figure Legend Snippet: The deficiency of DRD5 signaling increased colonic M1 macrophages but reduced M2 cells in the colon of DSS colitis mice. A Immunofluorescent labeling of F4/80 + , Arg1 + , and Inos + colonic macrophages of WT and DRD5 −/− mice on day 6 after DSS treatment (top). The cell numbers are quantified by Image pro plus (bottom) ( n = 3). Scale bars, 200 μm. B Flow cytometry analysis of colonic macrophages of WT and DRD5 −/− mice ( n = 5 mice per group) on day 6 after DSS treatment as indicated. Data are presented as representative plots (top) and summary graphs of quantified percentages (bottom). C Flow cytometry analysis of the CD45.2 + cells percentage in Arg1 + and CD86 + colonic macrophages of lethally irradiated mice reconstituted with DRD5 −/− CD45.2/WT CD45.1 bone marrow (1:1 ratio), and a control WT CD45.2/WT CD45.1 group (1:1 ratio) on day 6 after DSS treatment ( n = 3 mice per group). Data are presented as representative plots (left) and summary graphs of quantified percentages (right). Data are pooled from two or three independent experiments. Error bars show means ± SEM. * p

    Techniques Used: Mouse Assay, Labeling, Flow Cytometry, Irradiation

    The administration of DRD5 agonist attenuates the colitogenic phenotype of mice. A , B Age-matched male WT and DRD5 −/− mice ( n = 5 mice per group) were i.p. injected with D1-like agonist SKF-38393 at a dose of 10 mg/kg daily during DSS treatment. Weight changes ( A ) and disease activity index (DAI) ( B ) were monitored daily. C Gross morphology images of the DSS-induced colitis in WT mice or DRD5 −/− mice that were injected with DRD5 agonist, and colon length was measured on day 9. D Representative H E-staining section of colons and histology score of mice in ( A ). Scale bars, 4×, 500 μm; 20×, 200 μm. E The concentration of TNF-α, IL-6, and CCL2 in the serum of WT and DRD5 −/− mice treated with SKF38393. Data are pooled from three independent experiments. Error bars show means ± SEM. * P
    Figure Legend Snippet: The administration of DRD5 agonist attenuates the colitogenic phenotype of mice. A , B Age-matched male WT and DRD5 −/− mice ( n = 5 mice per group) were i.p. injected with D1-like agonist SKF-38393 at a dose of 10 mg/kg daily during DSS treatment. Weight changes ( A ) and disease activity index (DAI) ( B ) were monitored daily. C Gross morphology images of the DSS-induced colitis in WT mice or DRD5 −/− mice that were injected with DRD5 agonist, and colon length was measured on day 9. D Representative H E-staining section of colons and histology score of mice in ( A ). Scale bars, 4×, 500 μm; 20×, 200 μm. E The concentration of TNF-α, IL-6, and CCL2 in the serum of WT and DRD5 −/− mice treated with SKF38393. Data are pooled from three independent experiments. Error bars show means ± SEM. * P

    Techniques Used: Mouse Assay, Injection, Activity Assay, Staining, Concentration Assay

    DRD5 receptor is highly expressed in colonic macrophages. A RT-qPCR analysis of gene expression of DRD1 , DRD2 , DRD3 , DRD4 , and DRD5 in IECs (CD45 - Epcam + ) and LP cells (CD45 + Epcam - ) isolated from WT mice ( n = 3 mice per group). B RT-qPCR analysis of gene expression of DRD1 , DRD4 , and DRD5 in T cells, B cells, NK cells, DC cells, monocytes, neutrophils, macrophages, and ILCs cells isolated from the colonic LP of WT mice ( n = 3 mice per group). C Immunofluorescent labeling of TH (green), Cx3cr1(red), and DAPI (blue) in colon sections from Cx3cr1 reporter mice. The close proximity of macrophages with dopaminergic neurons is indicated by arrow. Scale bar, 30 µm. D Immunofluorescent labeling of DRD5 (green), F4/80 (red), and DAPI (blue) in colon sections from WT mice. The merging of DRD5 with F4/80 + in macrophages indicated by the arrowhead. Scale bar, 30 µm. E Immunofluorescent labeling of DRD5 (green), Arg1(red), and DAPI (blue) in colon sections from WT mice. The merging of DRD5 with Arg1 + in macrophages indicated by the arrowhead. Scale bar, 30 µm. F Immunofluorescent labeling of DRD5 (green), Inos (red), and DAPI (blue) in colon sections from WT mice. The merging of DRD5 with Inos + in macrophages indicated by the arrowhead. Scale bar, 30 µm. Data are pooled from three independent experiments ( A , B ). Error bars show means ± SEM. *** p
    Figure Legend Snippet: DRD5 receptor is highly expressed in colonic macrophages. A RT-qPCR analysis of gene expression of DRD1 , DRD2 , DRD3 , DRD4 , and DRD5 in IECs (CD45 - Epcam + ) and LP cells (CD45 + Epcam - ) isolated from WT mice ( n = 3 mice per group). B RT-qPCR analysis of gene expression of DRD1 , DRD4 , and DRD5 in T cells, B cells, NK cells, DC cells, monocytes, neutrophils, macrophages, and ILCs cells isolated from the colonic LP of WT mice ( n = 3 mice per group). C Immunofluorescent labeling of TH (green), Cx3cr1(red), and DAPI (blue) in colon sections from Cx3cr1 reporter mice. The close proximity of macrophages with dopaminergic neurons is indicated by arrow. Scale bar, 30 µm. D Immunofluorescent labeling of DRD5 (green), F4/80 (red), and DAPI (blue) in colon sections from WT mice. The merging of DRD5 with F4/80 + in macrophages indicated by the arrowhead. Scale bar, 30 µm. E Immunofluorescent labeling of DRD5 (green), Arg1(red), and DAPI (blue) in colon sections from WT mice. The merging of DRD5 with Arg1 + in macrophages indicated by the arrowhead. Scale bar, 30 µm. F Immunofluorescent labeling of DRD5 (green), Inos (red), and DAPI (blue) in colon sections from WT mice. The merging of DRD5 with Inos + in macrophages indicated by the arrowhead. Scale bar, 30 µm. Data are pooled from three independent experiments ( A , B ). Error bars show means ± SEM. *** p

    Techniques Used: Quantitative RT-PCR, Expressing, Isolation, Mouse Assay, Labeling

    DRD5 deficiency in immune cells exacerbates DSS-induced colitis. A , B Age-matched male WT and DRD5 − /− mice ( n = 9 mice per group) were given 2.5% DSS in their drinking water for 6 days and distilled water for three additional days before sacrifice. Weight changes ( A ) and disease activity index (DAI) ( B ) were monitored daily. C Gross morphology images of colons and colon lengths of WT and DRD5 − / − mice on day 9 after DSS treatment. D Representative H E-stained colonic sections and histology scores of WT and DRD5 − / − mice sampled on day 9 after DSS treatment. Scale bars, 4×, 500 μm; 20×, 200 μm. E The concentration of TNF-α, IL-6, and CCL2 in the serum of WT and DRD5 − /− mice were measured by ELISA. F , G Weight changes ( F ) and disease activity index (DAI) ( G ) after DSS-induced colitis in WT and DRD5 − / − mice ( n = 5 mice per group) adoptively transferred with WT or DRD5 −/− bone marrow cells. H Gross morphology images of colons and colon lengths of WT and DRD5 − /− mice adoptively transferred with WT or DRD5 − / − bone marrow cells on day 9 after DSS treatment. I Representative H E staining of colonic sections and histology scores of WT and DRD5 − /− mice adoptively transferred with WT or DRD5 − /− bone marrow cells on day 9 after DSS treatment. Scale bars, 4×, 500 μm; 20×, 200 μm. J The concentration of TNF-α, IL-6, and CCL2 in the serum of WT and DRD5 −/ − mice ( n = 5 mice per group) adoptively transferred with WT or DRD5 − /− bone marrow cells after DSS treatment. Data are pooled from three independent experiments. Error bars show means ± SEM. * P
    Figure Legend Snippet: DRD5 deficiency in immune cells exacerbates DSS-induced colitis. A , B Age-matched male WT and DRD5 − /− mice ( n = 9 mice per group) were given 2.5% DSS in their drinking water for 6 days and distilled water for three additional days before sacrifice. Weight changes ( A ) and disease activity index (DAI) ( B ) were monitored daily. C Gross morphology images of colons and colon lengths of WT and DRD5 − / − mice on day 9 after DSS treatment. D Representative H E-stained colonic sections and histology scores of WT and DRD5 − / − mice sampled on day 9 after DSS treatment. Scale bars, 4×, 500 μm; 20×, 200 μm. E The concentration of TNF-α, IL-6, and CCL2 in the serum of WT and DRD5 − /− mice were measured by ELISA. F , G Weight changes ( F ) and disease activity index (DAI) ( G ) after DSS-induced colitis in WT and DRD5 − / − mice ( n = 5 mice per group) adoptively transferred with WT or DRD5 −/− bone marrow cells. H Gross morphology images of colons and colon lengths of WT and DRD5 − /− mice adoptively transferred with WT or DRD5 − / − bone marrow cells on day 9 after DSS treatment. I Representative H E staining of colonic sections and histology scores of WT and DRD5 − /− mice adoptively transferred with WT or DRD5 − /− bone marrow cells on day 9 after DSS treatment. Scale bars, 4×, 500 μm; 20×, 200 μm. J The concentration of TNF-α, IL-6, and CCL2 in the serum of WT and DRD5 −/ − mice ( n = 5 mice per group) adoptively transferred with WT or DRD5 − /− bone marrow cells after DSS treatment. Data are pooled from three independent experiments. Error bars show means ± SEM. * P

    Techniques Used: Mouse Assay, Activity Assay, Staining, Concentration Assay, Enzyme-linked Immunosorbent Assay

    DA-DRD5 signaling can inhibit M1 polarization by negatively regulating NF-κB signaling and promote M2 polarization through the activation of the CREB pathway. A Principal component (PC) analysis of transcriptional clustered architecture in M1-stimulated WT and DRD5 −/− BMDMs untreated or treated with DA (20 μM) ( n = 3). B Gene set enrichment analysis (GSEA) analysis of the prominent down-regulated pathways in M1-stimulated WT BMDMs untreated or treated with DA (left) or M1-stimulated WT and DRD5 −/− BMDMs treated with DA (right). C Immunoblot analysis of phosphorylated (p−), and total IKKα/β, IκBα in lysates of LPS/IFN-γ-stimulated WT and DRD5 −/− BMDMs untreated or treated with DA (20 μM) as indicated. D Heat map showing the expression of NF-κB-mediated M1 genes in LPS/IFN-γ-stimulated WT and DRD5 −/− BMDMs untreated or treated with DA. E PC analysis of gene expression cluster in M2-stimulated WT and DRD5 −/− BMDMs untreated or treated with DA (20 μM) ( n = 3). F GSEA analysis of the prominent upregulated pathways in M2-stimulated WT BMDMs untreated or treated with DA (left) or M2-stimulated WT and DRD5 −/− BMDMs treated with DA (right). G Immunoblot analysis of phosphorylated (p−), and total CREB, PKAc in lysates of IL-4/IL-13-stimulated WT and DRD5 −/− BMDMs untreated or treated with DA (20 μM) as indicated. H Heat map showing the expression of genes associated with M2 macrophage in IL-4/IL-13-stimulated WT and DRD5 −/− BMDMs untreated or treated with DA. Data are representative of three independent experiments ( C , G ).
    Figure Legend Snippet: DA-DRD5 signaling can inhibit M1 polarization by negatively regulating NF-κB signaling and promote M2 polarization through the activation of the CREB pathway. A Principal component (PC) analysis of transcriptional clustered architecture in M1-stimulated WT and DRD5 −/− BMDMs untreated or treated with DA (20 μM) ( n = 3). B Gene set enrichment analysis (GSEA) analysis of the prominent down-regulated pathways in M1-stimulated WT BMDMs untreated or treated with DA (left) or M1-stimulated WT and DRD5 −/− BMDMs treated with DA (right). C Immunoblot analysis of phosphorylated (p−), and total IKKα/β, IκBα in lysates of LPS/IFN-γ-stimulated WT and DRD5 −/− BMDMs untreated or treated with DA (20 μM) as indicated. D Heat map showing the expression of NF-κB-mediated M1 genes in LPS/IFN-γ-stimulated WT and DRD5 −/− BMDMs untreated or treated with DA. E PC analysis of gene expression cluster in M2-stimulated WT and DRD5 −/− BMDMs untreated or treated with DA (20 μM) ( n = 3). F GSEA analysis of the prominent upregulated pathways in M2-stimulated WT BMDMs untreated or treated with DA (left) or M2-stimulated WT and DRD5 −/− BMDMs treated with DA (right). G Immunoblot analysis of phosphorylated (p−), and total CREB, PKAc in lysates of IL-4/IL-13-stimulated WT and DRD5 −/− BMDMs untreated or treated with DA (20 μM) as indicated. H Heat map showing the expression of genes associated with M2 macrophage in IL-4/IL-13-stimulated WT and DRD5 −/− BMDMs untreated or treated with DA. Data are representative of three independent experiments ( C , G ).

    Techniques Used: Activation Assay, Expressing

    The deficiency of DA-DRD5 signaling inhibits M1 but enhances M2 macrophage polarization in vitro. A RT-qPCR analysis of Inos and Tnf mRNA expression in WT BMDMs treated with various doses of DA and stimulated with LPS/IFN-γ (M1) for 12 h. B ELISA analysis of TNF-α in supernatants from WT BMDMs treated with various doses of DA and stimulated with LPS/IFN-γ (M1) for 12 h. C Flow cytometry analysis of M1 marker CD86 of WT BMDMs treated with DA (20 μM) and stimulated with LPS/IFN-γ (M1) for 12 h. Representative overlaid flow cytometry histograms showing mean fluorescence intensity (MFI) (top) and quantified data from three independent experiments ( n = 3 per group) (bottom). D Immunoblot analysis of Inos in lysates of WT BMDMs treated with DA (20 μM) and stimulated with LPS/IFN-γ (M1) for 12 h. E RT-qPCR analysis of Inos and Tnf mRNA expression in WT and DRD5 −/− BMDMs treated with DA (20 μM) and stimulated with LPS/IFN-γ (M1) for 12 h. F ELISA analysis of TNF-α in supernatants from WT and DRD5 −/− BMDMs treated with DA (20 μM) and stimulated with LPS/IFN-γ (M1) for 12 h. G Flow cytometry analysis of M1 marker CD86 of WT and DRD5 −/− BMDMs treated with DA (20μM) and stimulated with LPS/IFN-γ (M1) for 12 h. Representative overlaid flow cytometry histograms showing mean fluorescence intensity (MFI) (left) and quantified data from three independent experiments ( n = 3 per group) (right). H Immunoblot analysis of Inos in lysates of WT and DRD5 −/− BMDMs treated with DA (20 μM) and stimulated with LPS/IFN-γ (M1) for 12 h. I RT-qPCR analysis of Arg1 and Mrc1 mRNA expression in WT BMDMs treated with various doses of DA and stimulated with IL-4/IL-13 (M2) for 12 h. J Flow cytometry analysis of M2 marker CD206 of WT BMDMs treated with DA (20 μM) and stimulated with IL-4/IL-13 (M2) for 12 h. Representative overlaid flow cytometry histograms showing mean fluorescence intensity (MFI) (left) and quantified data from three independent experiments ( n = 3 per group) (right). K Immunoblot analysis of Arg1 in lysates of WT BMDMs treated with DA (20 μM) and stimulated with M2 for 12 h. L RT-qPCR analysis of Arg1 and Mrc1 mRNA expression in WT and DRD5 −/− BMDMs treated with DA (20 μM) and stimulated with IL-4/IL-13 (M2) for 12 h. M Flow cytometry analysis of M2 marker CD206 of WT and DRD5 −/− BMDMs treated with DA (20 μM) and stimulated with IL-4/IL-13 (M2) for 12 h. Representative overlaid flow cytometry histograms showing mean fluorescence intensity (MFI) (left) and quantified data from three independent experiments ( n = 3 per group) (right). N Immunoblot analysis of Arg1 in lysates of WT and DRD5 −/− BMDMs treated with DA (20 μM) and stimulated with IL-4/IL-13 (M2) for 12 h. Data are pooled from three independent experiments. Error bars show means ± SEM. * p
    Figure Legend Snippet: The deficiency of DA-DRD5 signaling inhibits M1 but enhances M2 macrophage polarization in vitro. A RT-qPCR analysis of Inos and Tnf mRNA expression in WT BMDMs treated with various doses of DA and stimulated with LPS/IFN-γ (M1) for 12 h. B ELISA analysis of TNF-α in supernatants from WT BMDMs treated with various doses of DA and stimulated with LPS/IFN-γ (M1) for 12 h. C Flow cytometry analysis of M1 marker CD86 of WT BMDMs treated with DA (20 μM) and stimulated with LPS/IFN-γ (M1) for 12 h. Representative overlaid flow cytometry histograms showing mean fluorescence intensity (MFI) (top) and quantified data from three independent experiments ( n = 3 per group) (bottom). D Immunoblot analysis of Inos in lysates of WT BMDMs treated with DA (20 μM) and stimulated with LPS/IFN-γ (M1) for 12 h. E RT-qPCR analysis of Inos and Tnf mRNA expression in WT and DRD5 −/− BMDMs treated with DA (20 μM) and stimulated with LPS/IFN-γ (M1) for 12 h. F ELISA analysis of TNF-α in supernatants from WT and DRD5 −/− BMDMs treated with DA (20 μM) and stimulated with LPS/IFN-γ (M1) for 12 h. G Flow cytometry analysis of M1 marker CD86 of WT and DRD5 −/− BMDMs treated with DA (20μM) and stimulated with LPS/IFN-γ (M1) for 12 h. Representative overlaid flow cytometry histograms showing mean fluorescence intensity (MFI) (left) and quantified data from three independent experiments ( n = 3 per group) (right). H Immunoblot analysis of Inos in lysates of WT and DRD5 −/− BMDMs treated with DA (20 μM) and stimulated with LPS/IFN-γ (M1) for 12 h. I RT-qPCR analysis of Arg1 and Mrc1 mRNA expression in WT BMDMs treated with various doses of DA and stimulated with IL-4/IL-13 (M2) for 12 h. J Flow cytometry analysis of M2 marker CD206 of WT BMDMs treated with DA (20 μM) and stimulated with IL-4/IL-13 (M2) for 12 h. Representative overlaid flow cytometry histograms showing mean fluorescence intensity (MFI) (left) and quantified data from three independent experiments ( n = 3 per group) (right). K Immunoblot analysis of Arg1 in lysates of WT BMDMs treated with DA (20 μM) and stimulated with M2 for 12 h. L RT-qPCR analysis of Arg1 and Mrc1 mRNA expression in WT and DRD5 −/− BMDMs treated with DA (20 μM) and stimulated with IL-4/IL-13 (M2) for 12 h. M Flow cytometry analysis of M2 marker CD206 of WT and DRD5 −/− BMDMs treated with DA (20 μM) and stimulated with IL-4/IL-13 (M2) for 12 h. Representative overlaid flow cytometry histograms showing mean fluorescence intensity (MFI) (left) and quantified data from three independent experiments ( n = 3 per group) (right). N Immunoblot analysis of Arg1 in lysates of WT and DRD5 −/− BMDMs treated with DA (20 μM) and stimulated with IL-4/IL-13 (M2) for 12 h. Data are pooled from three independent experiments. Error bars show means ± SEM. * p

    Techniques Used: In Vitro, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Marker, Fluorescence

    2) Product Images from "A GABAergic cell type in the lateral habenula links hypothalamic homeostatic and midbrain motivation circuits with sex steroid signaling"

    Article Title: A GABAergic cell type in the lateral habenula links hypothalamic homeostatic and midbrain motivation circuits with sex steroid signaling

    Journal: Translational Psychiatry

    doi: 10.1038/s41398-018-0099-5

    GABAergic estrogen-receptive neurons (GERNs) localization, mRNA expression by ISH and somatic receptor expression by IHC. a  Serial coronal sections showing the ERα immunolabelling at the Bregma rostro-caudal coordinates (numbers in mm under the photomicrographs). Boxed area in A 6  at higher magnification showing the exclusive nuclear labeling pattern. The bold numbered levels (A 1  and A 8 ), were chosen to show that no positive labeling was found in either rostral or caudal directions. A 9 : a horizontal view of the distribution of estrogen-receptive cells was symbolized by the red oval. A 10 : sagittal view of rat brain atlas, modified from Paxinos    Watson  86 , at lat. 0.90 mm, where lateral habenula (LHb) is symbolized with a gray shade and A9 plane was symbolized with a horizontal line.  b  In situ hybridization using multiple RNAscope methods. B 1 : multiplex fluorescence method, Esr1, gene that encodes ERα (red punctuated labeling) co-expressed with Slc32a1, gene that encodes VGAT (green punctuated labeling); arrows indicate the double-labeled cells; B 2 : duplex method, Esr1 (red punctuated labeling) co-localization with Slc17a6, gene that encodes VGLUT2 (green punctuated labeling); arrows indicate the double-labeled cells; B 3 : with duplex method, Slc32a1 encoding VGAT (red punctuated labeling) shows complete co-localization with Slc17a6 encoding VGLUT2 (green punctuated labeling); arrows indicate the double-labeled cells. Inset of B 3 , Slc32a1 expression in a sexually active (SA) rat LHb, Br. −3.72 mm (brown labeling, single chromogenic-Brown method-RNAscope). *Note the similarity with ERα expression in A 6 .  c  Indirect immunohistochemistry showing the GABAergic nature of the ERα+ neurons (red) in a SA rat brain. The GABA antibody (green, Sigma, A0310) produced characteristic surface labeling (C 1 , C 2 : the strip-like image was produced by Vibratome slicing leaving the brain section with an uneven surface).  d  ERα+ neurons co-express receptor/receptor subtypes for vasopressin, orexin, dopamine, and serotonin. D 1 : In situ hybridization using RNAscope-multiplex method targeting Esr1 (red dots), Slc32a1 (white dots) and Hcrtr2, gene encodes the orexin receptor 2 (green dots). D 2 , D 3 : Indirect immunofluorescence reactions, showing that the ERα-IR cells co-expressed vasopressin receptor V1a (inset showing the V1a antibody labeling pattern in temporal hippocampus CA2 cells body layer. See also SI Fig.   6  for more information about this antibody), dopamine receptor D5R (also called D1Rb) and serotonin receptor 5-HTr2a, respectively. Scale bars: A 5  and  b : 500 µm and rest: 10 µm
    Figure Legend Snippet: GABAergic estrogen-receptive neurons (GERNs) localization, mRNA expression by ISH and somatic receptor expression by IHC. a Serial coronal sections showing the ERα immunolabelling at the Bregma rostro-caudal coordinates (numbers in mm under the photomicrographs). Boxed area in A 6 at higher magnification showing the exclusive nuclear labeling pattern. The bold numbered levels (A 1 and A 8 ), were chosen to show that no positive labeling was found in either rostral or caudal directions. A 9 : a horizontal view of the distribution of estrogen-receptive cells was symbolized by the red oval. A 10 : sagittal view of rat brain atlas, modified from Paxinos Watson 86 , at lat. 0.90 mm, where lateral habenula (LHb) is symbolized with a gray shade and A9 plane was symbolized with a horizontal line. b In situ hybridization using multiple RNAscope methods. B 1 : multiplex fluorescence method, Esr1, gene that encodes ERα (red punctuated labeling) co-expressed with Slc32a1, gene that encodes VGAT (green punctuated labeling); arrows indicate the double-labeled cells; B 2 : duplex method, Esr1 (red punctuated labeling) co-localization with Slc17a6, gene that encodes VGLUT2 (green punctuated labeling); arrows indicate the double-labeled cells; B 3 : with duplex method, Slc32a1 encoding VGAT (red punctuated labeling) shows complete co-localization with Slc17a6 encoding VGLUT2 (green punctuated labeling); arrows indicate the double-labeled cells. Inset of B 3 , Slc32a1 expression in a sexually active (SA) rat LHb, Br. −3.72 mm (brown labeling, single chromogenic-Brown method-RNAscope). *Note the similarity with ERα expression in A 6 . c Indirect immunohistochemistry showing the GABAergic nature of the ERα+ neurons (red) in a SA rat brain. The GABA antibody (green, Sigma, A0310) produced characteristic surface labeling (C 1 , C 2 : the strip-like image was produced by Vibratome slicing leaving the brain section with an uneven surface). d ERα+ neurons co-express receptor/receptor subtypes for vasopressin, orexin, dopamine, and serotonin. D 1 : In situ hybridization using RNAscope-multiplex method targeting Esr1 (red dots), Slc32a1 (white dots) and Hcrtr2, gene encodes the orexin receptor 2 (green dots). D 2 , D 3 : Indirect immunofluorescence reactions, showing that the ERα-IR cells co-expressed vasopressin receptor V1a (inset showing the V1a antibody labeling pattern in temporal hippocampus CA2 cells body layer. See also SI Fig. 6 for more information about this antibody), dopamine receptor D5R (also called D1Rb) and serotonin receptor 5-HTr2a, respectively. Scale bars: A 5 and b : 500 µm and rest: 10 µm

    Techniques Used: Expressing, In Situ Hybridization, Immunohistochemistry, Labeling, Modification, Multiplex Assay, Fluorescence, Produced, Stripping Membranes, Immunofluorescence, Antibody Labeling

    3) Product Images from "Differential Expression of Dopamine D5 Receptors across Neuronal Subtypes in Macaque Frontal Eye Field"

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

    Journal: Frontiers in Neural Circuits

    doi: 10.3389/fncir.2018.00012

    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.
    Figure Legend 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.

    Techniques Used:

    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 ††† .
    Figure Legend 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 ††† .

    Techniques Used:

    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).
    Figure Legend 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).

    Techniques Used: Marker

    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.
    Figure Legend 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.

    Techniques Used: Expressing, Staining

    4) Product Images from "Differential Expression of Dopamine D5 Receptors across Neuronal Subtypes in Macaque Frontal Eye Field"

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

    Journal: Frontiers in Neural Circuits

    doi: 10.3389/fncir.2018.00012

    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.
    Figure Legend 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.

    Techniques Used: Expressing, Staining

    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.
    Figure Legend 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.

    Techniques Used:

    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 ††† .
    Figure Legend 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 ††† .

    Techniques Used:

    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).
    Figure Legend 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).

    Techniques Used: Marker

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    Alomone Labs anti d5 dopamine receptor extracellular antibody
    The deficiency of <t>DRD5</t> signaling increased colonic M1 macrophages but reduced M2 cells in the colon of DSS colitis mice. A Immunofluorescent labeling of F4/80 + , Arg1 + , and Inos + colonic macrophages of WT and DRD5 −/− mice on day 6 after DSS treatment (top). The cell numbers are quantified by Image pro plus (bottom) ( n = 3). Scale bars, 200 μm. B Flow cytometry analysis of colonic macrophages of WT and DRD5 −/− mice ( n = 5 mice per group) on day 6 after DSS treatment as indicated. Data are presented as representative plots (top) and summary graphs of quantified percentages (bottom). C Flow cytometry analysis of the CD45.2 + cells percentage in Arg1 + and CD86 + colonic macrophages of lethally irradiated mice reconstituted with DRD5 −/− CD45.2/WT CD45.1 bone marrow (1:1 ratio), and a control WT CD45.2/WT CD45.1 group (1:1 ratio) on day 6 after DSS treatment ( n = 3 mice per group). Data are presented as representative plots (left) and summary graphs of quantified percentages (right). Data are pooled from two or three independent experiments. Error bars show means ± SEM. * p
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    The deficiency of DRD5 signaling increased colonic M1 macrophages but reduced M2 cells in the colon of DSS colitis mice. A Immunofluorescent labeling of F4/80 + , Arg1 + , and Inos + colonic macrophages of WT and DRD5 −/− mice on day 6 after DSS treatment (top). The cell numbers are quantified by Image pro plus (bottom) ( n = 3). Scale bars, 200 μm. B Flow cytometry analysis of colonic macrophages of WT and DRD5 −/− mice ( n = 5 mice per group) on day 6 after DSS treatment as indicated. Data are presented as representative plots (top) and summary graphs of quantified percentages (bottom). C Flow cytometry analysis of the CD45.2 + cells percentage in Arg1 + and CD86 + colonic macrophages of lethally irradiated mice reconstituted with DRD5 −/− CD45.2/WT CD45.1 bone marrow (1:1 ratio), and a control WT CD45.2/WT CD45.1 group (1:1 ratio) on day 6 after DSS treatment ( n = 3 mice per group). Data are presented as representative plots (left) and summary graphs of quantified percentages (right). Data are pooled from two or three independent experiments. Error bars show means ± SEM. * p

    Journal: Cell Death & Disease

    Article Title: DA-DRD5 signaling controls colitis by regulating colonic M1/M2 macrophage polarization

    doi: 10.1038/s41419-021-03778-6

    Figure Lengend Snippet: The deficiency of DRD5 signaling increased colonic M1 macrophages but reduced M2 cells in the colon of DSS colitis mice. A Immunofluorescent labeling of F4/80 + , Arg1 + , and Inos + colonic macrophages of WT and DRD5 −/− mice on day 6 after DSS treatment (top). The cell numbers are quantified by Image pro plus (bottom) ( n = 3). Scale bars, 200 μm. B Flow cytometry analysis of colonic macrophages of WT and DRD5 −/− mice ( n = 5 mice per group) on day 6 after DSS treatment as indicated. Data are presented as representative plots (top) and summary graphs of quantified percentages (bottom). C Flow cytometry analysis of the CD45.2 + cells percentage in Arg1 + and CD86 + colonic macrophages of lethally irradiated mice reconstituted with DRD5 −/− CD45.2/WT CD45.1 bone marrow (1:1 ratio), and a control WT CD45.2/WT CD45.1 group (1:1 ratio) on day 6 after DSS treatment ( n = 3 mice per group). Data are presented as representative plots (left) and summary graphs of quantified percentages (right). Data are pooled from two or three independent experiments. Error bars show means ± SEM. * p

    Article Snippet: Anti-PKA C-α (55388-1-AP) and anti-Drd5 (20310-1-AP/ADR-005) antibodies were from Proteintech/Alomone.

    Techniques: Mouse Assay, Labeling, Flow Cytometry, Irradiation

    The administration of DRD5 agonist attenuates the colitogenic phenotype of mice. A , B Age-matched male WT and DRD5 −/− mice ( n = 5 mice per group) were i.p. injected with D1-like agonist SKF-38393 at a dose of 10 mg/kg daily during DSS treatment. Weight changes ( A ) and disease activity index (DAI) ( B ) were monitored daily. C Gross morphology images of the DSS-induced colitis in WT mice or DRD5 −/− mice that were injected with DRD5 agonist, and colon length was measured on day 9. D Representative H E-staining section of colons and histology score of mice in ( A ). Scale bars, 4×, 500 μm; 20×, 200 μm. E The concentration of TNF-α, IL-6, and CCL2 in the serum of WT and DRD5 −/− mice treated with SKF38393. Data are pooled from three independent experiments. Error bars show means ± SEM. * P

    Journal: Cell Death & Disease

    Article Title: DA-DRD5 signaling controls colitis by regulating colonic M1/M2 macrophage polarization

    doi: 10.1038/s41419-021-03778-6

    Figure Lengend Snippet: The administration of DRD5 agonist attenuates the colitogenic phenotype of mice. A , B Age-matched male WT and DRD5 −/− mice ( n = 5 mice per group) were i.p. injected with D1-like agonist SKF-38393 at a dose of 10 mg/kg daily during DSS treatment. Weight changes ( A ) and disease activity index (DAI) ( B ) were monitored daily. C Gross morphology images of the DSS-induced colitis in WT mice or DRD5 −/− mice that were injected with DRD5 agonist, and colon length was measured on day 9. D Representative H E-staining section of colons and histology score of mice in ( A ). Scale bars, 4×, 500 μm; 20×, 200 μm. E The concentration of TNF-α, IL-6, and CCL2 in the serum of WT and DRD5 −/− mice treated with SKF38393. Data are pooled from three independent experiments. Error bars show means ± SEM. * P

    Article Snippet: Anti-PKA C-α (55388-1-AP) and anti-Drd5 (20310-1-AP/ADR-005) antibodies were from Proteintech/Alomone.

    Techniques: Mouse Assay, Injection, Activity Assay, Staining, Concentration Assay

    DRD5 receptor is highly expressed in colonic macrophages. A RT-qPCR analysis of gene expression of DRD1 , DRD2 , DRD3 , DRD4 , and DRD5 in IECs (CD45 - Epcam + ) and LP cells (CD45 + Epcam - ) isolated from WT mice ( n = 3 mice per group). B RT-qPCR analysis of gene expression of DRD1 , DRD4 , and DRD5 in T cells, B cells, NK cells, DC cells, monocytes, neutrophils, macrophages, and ILCs cells isolated from the colonic LP of WT mice ( n = 3 mice per group). C Immunofluorescent labeling of TH (green), Cx3cr1(red), and DAPI (blue) in colon sections from Cx3cr1 reporter mice. The close proximity of macrophages with dopaminergic neurons is indicated by arrow. Scale bar, 30 µm. D Immunofluorescent labeling of DRD5 (green), F4/80 (red), and DAPI (blue) in colon sections from WT mice. The merging of DRD5 with F4/80 + in macrophages indicated by the arrowhead. Scale bar, 30 µm. E Immunofluorescent labeling of DRD5 (green), Arg1(red), and DAPI (blue) in colon sections from WT mice. The merging of DRD5 with Arg1 + in macrophages indicated by the arrowhead. Scale bar, 30 µm. F Immunofluorescent labeling of DRD5 (green), Inos (red), and DAPI (blue) in colon sections from WT mice. The merging of DRD5 with Inos + in macrophages indicated by the arrowhead. Scale bar, 30 µm. Data are pooled from three independent experiments ( A , B ). Error bars show means ± SEM. *** p

    Journal: Cell Death & Disease

    Article Title: DA-DRD5 signaling controls colitis by regulating colonic M1/M2 macrophage polarization

    doi: 10.1038/s41419-021-03778-6

    Figure Lengend Snippet: DRD5 receptor is highly expressed in colonic macrophages. A RT-qPCR analysis of gene expression of DRD1 , DRD2 , DRD3 , DRD4 , and DRD5 in IECs (CD45 - Epcam + ) and LP cells (CD45 + Epcam - ) isolated from WT mice ( n = 3 mice per group). B RT-qPCR analysis of gene expression of DRD1 , DRD4 , and DRD5 in T cells, B cells, NK cells, DC cells, monocytes, neutrophils, macrophages, and ILCs cells isolated from the colonic LP of WT mice ( n = 3 mice per group). C Immunofluorescent labeling of TH (green), Cx3cr1(red), and DAPI (blue) in colon sections from Cx3cr1 reporter mice. The close proximity of macrophages with dopaminergic neurons is indicated by arrow. Scale bar, 30 µm. D Immunofluorescent labeling of DRD5 (green), F4/80 (red), and DAPI (blue) in colon sections from WT mice. The merging of DRD5 with F4/80 + in macrophages indicated by the arrowhead. Scale bar, 30 µm. E Immunofluorescent labeling of DRD5 (green), Arg1(red), and DAPI (blue) in colon sections from WT mice. The merging of DRD5 with Arg1 + in macrophages indicated by the arrowhead. Scale bar, 30 µm. F Immunofluorescent labeling of DRD5 (green), Inos (red), and DAPI (blue) in colon sections from WT mice. The merging of DRD5 with Inos + in macrophages indicated by the arrowhead. Scale bar, 30 µm. Data are pooled from three independent experiments ( A , B ). Error bars show means ± SEM. *** p

    Article Snippet: Anti-PKA C-α (55388-1-AP) and anti-Drd5 (20310-1-AP/ADR-005) antibodies were from Proteintech/Alomone.

    Techniques: Quantitative RT-PCR, Expressing, Isolation, Mouse Assay, Labeling

    DRD5 deficiency in immune cells exacerbates DSS-induced colitis. A , B Age-matched male WT and DRD5 − /− mice ( n = 9 mice per group) were given 2.5% DSS in their drinking water for 6 days and distilled water for three additional days before sacrifice. Weight changes ( A ) and disease activity index (DAI) ( B ) were monitored daily. C Gross morphology images of colons and colon lengths of WT and DRD5 − / − mice on day 9 after DSS treatment. D Representative H E-stained colonic sections and histology scores of WT and DRD5 − / − mice sampled on day 9 after DSS treatment. Scale bars, 4×, 500 μm; 20×, 200 μm. E The concentration of TNF-α, IL-6, and CCL2 in the serum of WT and DRD5 − /− mice were measured by ELISA. F , G Weight changes ( F ) and disease activity index (DAI) ( G ) after DSS-induced colitis in WT and DRD5 − / − mice ( n = 5 mice per group) adoptively transferred with WT or DRD5 −/− bone marrow cells. H Gross morphology images of colons and colon lengths of WT and DRD5 − /− mice adoptively transferred with WT or DRD5 − / − bone marrow cells on day 9 after DSS treatment. I Representative H E staining of colonic sections and histology scores of WT and DRD5 − /− mice adoptively transferred with WT or DRD5 − /− bone marrow cells on day 9 after DSS treatment. Scale bars, 4×, 500 μm; 20×, 200 μm. J The concentration of TNF-α, IL-6, and CCL2 in the serum of WT and DRD5 −/ − mice ( n = 5 mice per group) adoptively transferred with WT or DRD5 − /− bone marrow cells after DSS treatment. Data are pooled from three independent experiments. Error bars show means ± SEM. * P

    Journal: Cell Death & Disease

    Article Title: DA-DRD5 signaling controls colitis by regulating colonic M1/M2 macrophage polarization

    doi: 10.1038/s41419-021-03778-6

    Figure Lengend Snippet: DRD5 deficiency in immune cells exacerbates DSS-induced colitis. A , B Age-matched male WT and DRD5 − /− mice ( n = 9 mice per group) were given 2.5% DSS in their drinking water for 6 days and distilled water for three additional days before sacrifice. Weight changes ( A ) and disease activity index (DAI) ( B ) were monitored daily. C Gross morphology images of colons and colon lengths of WT and DRD5 − / − mice on day 9 after DSS treatment. D Representative H E-stained colonic sections and histology scores of WT and DRD5 − / − mice sampled on day 9 after DSS treatment. Scale bars, 4×, 500 μm; 20×, 200 μm. E The concentration of TNF-α, IL-6, and CCL2 in the serum of WT and DRD5 − /− mice were measured by ELISA. F , G Weight changes ( F ) and disease activity index (DAI) ( G ) after DSS-induced colitis in WT and DRD5 − / − mice ( n = 5 mice per group) adoptively transferred with WT or DRD5 −/− bone marrow cells. H Gross morphology images of colons and colon lengths of WT and DRD5 − /− mice adoptively transferred with WT or DRD5 − / − bone marrow cells on day 9 after DSS treatment. I Representative H E staining of colonic sections and histology scores of WT and DRD5 − /− mice adoptively transferred with WT or DRD5 − /− bone marrow cells on day 9 after DSS treatment. Scale bars, 4×, 500 μm; 20×, 200 μm. J The concentration of TNF-α, IL-6, and CCL2 in the serum of WT and DRD5 −/ − mice ( n = 5 mice per group) adoptively transferred with WT or DRD5 − /− bone marrow cells after DSS treatment. Data are pooled from three independent experiments. Error bars show means ± SEM. * P

    Article Snippet: Anti-PKA C-α (55388-1-AP) and anti-Drd5 (20310-1-AP/ADR-005) antibodies were from Proteintech/Alomone.

    Techniques: Mouse Assay, Activity Assay, Staining, Concentration Assay, Enzyme-linked Immunosorbent Assay

    DA-DRD5 signaling can inhibit M1 polarization by negatively regulating NF-κB signaling and promote M2 polarization through the activation of the CREB pathway. A Principal component (PC) analysis of transcriptional clustered architecture in M1-stimulated WT and DRD5 −/− BMDMs untreated or treated with DA (20 μM) ( n = 3). B Gene set enrichment analysis (GSEA) analysis of the prominent down-regulated pathways in M1-stimulated WT BMDMs untreated or treated with DA (left) or M1-stimulated WT and DRD5 −/− BMDMs treated with DA (right). C Immunoblot analysis of phosphorylated (p−), and total IKKα/β, IκBα in lysates of LPS/IFN-γ-stimulated WT and DRD5 −/− BMDMs untreated or treated with DA (20 μM) as indicated. D Heat map showing the expression of NF-κB-mediated M1 genes in LPS/IFN-γ-stimulated WT and DRD5 −/− BMDMs untreated or treated with DA. E PC analysis of gene expression cluster in M2-stimulated WT and DRD5 −/− BMDMs untreated or treated with DA (20 μM) ( n = 3). F GSEA analysis of the prominent upregulated pathways in M2-stimulated WT BMDMs untreated or treated with DA (left) or M2-stimulated WT and DRD5 −/− BMDMs treated with DA (right). G Immunoblot analysis of phosphorylated (p−), and total CREB, PKAc in lysates of IL-4/IL-13-stimulated WT and DRD5 −/− BMDMs untreated or treated with DA (20 μM) as indicated. H Heat map showing the expression of genes associated with M2 macrophage in IL-4/IL-13-stimulated WT and DRD5 −/− BMDMs untreated or treated with DA. Data are representative of three independent experiments ( C , G ).

    Journal: Cell Death & Disease

    Article Title: DA-DRD5 signaling controls colitis by regulating colonic M1/M2 macrophage polarization

    doi: 10.1038/s41419-021-03778-6

    Figure Lengend Snippet: DA-DRD5 signaling can inhibit M1 polarization by negatively regulating NF-κB signaling and promote M2 polarization through the activation of the CREB pathway. A Principal component (PC) analysis of transcriptional clustered architecture in M1-stimulated WT and DRD5 −/− BMDMs untreated or treated with DA (20 μM) ( n = 3). B Gene set enrichment analysis (GSEA) analysis of the prominent down-regulated pathways in M1-stimulated WT BMDMs untreated or treated with DA (left) or M1-stimulated WT and DRD5 −/− BMDMs treated with DA (right). C Immunoblot analysis of phosphorylated (p−), and total IKKα/β, IκBα in lysates of LPS/IFN-γ-stimulated WT and DRD5 −/− BMDMs untreated or treated with DA (20 μM) as indicated. D Heat map showing the expression of NF-κB-mediated M1 genes in LPS/IFN-γ-stimulated WT and DRD5 −/− BMDMs untreated or treated with DA. E PC analysis of gene expression cluster in M2-stimulated WT and DRD5 −/− BMDMs untreated or treated with DA (20 μM) ( n = 3). F GSEA analysis of the prominent upregulated pathways in M2-stimulated WT BMDMs untreated or treated with DA (left) or M2-stimulated WT and DRD5 −/− BMDMs treated with DA (right). G Immunoblot analysis of phosphorylated (p−), and total CREB, PKAc in lysates of IL-4/IL-13-stimulated WT and DRD5 −/− BMDMs untreated or treated with DA (20 μM) as indicated. H Heat map showing the expression of genes associated with M2 macrophage in IL-4/IL-13-stimulated WT and DRD5 −/− BMDMs untreated or treated with DA. Data are representative of three independent experiments ( C , G ).

    Article Snippet: Anti-PKA C-α (55388-1-AP) and anti-Drd5 (20310-1-AP/ADR-005) antibodies were from Proteintech/Alomone.

    Techniques: Activation Assay, Expressing

    The deficiency of DA-DRD5 signaling inhibits M1 but enhances M2 macrophage polarization in vitro. A RT-qPCR analysis of Inos and Tnf mRNA expression in WT BMDMs treated with various doses of DA and stimulated with LPS/IFN-γ (M1) for 12 h. B ELISA analysis of TNF-α in supernatants from WT BMDMs treated with various doses of DA and stimulated with LPS/IFN-γ (M1) for 12 h. C Flow cytometry analysis of M1 marker CD86 of WT BMDMs treated with DA (20 μM) and stimulated with LPS/IFN-γ (M1) for 12 h. Representative overlaid flow cytometry histograms showing mean fluorescence intensity (MFI) (top) and quantified data from three independent experiments ( n = 3 per group) (bottom). D Immunoblot analysis of Inos in lysates of WT BMDMs treated with DA (20 μM) and stimulated with LPS/IFN-γ (M1) for 12 h. E RT-qPCR analysis of Inos and Tnf mRNA expression in WT and DRD5 −/− BMDMs treated with DA (20 μM) and stimulated with LPS/IFN-γ (M1) for 12 h. F ELISA analysis of TNF-α in supernatants from WT and DRD5 −/− BMDMs treated with DA (20 μM) and stimulated with LPS/IFN-γ (M1) for 12 h. G Flow cytometry analysis of M1 marker CD86 of WT and DRD5 −/− BMDMs treated with DA (20μM) and stimulated with LPS/IFN-γ (M1) for 12 h. Representative overlaid flow cytometry histograms showing mean fluorescence intensity (MFI) (left) and quantified data from three independent experiments ( n = 3 per group) (right). H Immunoblot analysis of Inos in lysates of WT and DRD5 −/− BMDMs treated with DA (20 μM) and stimulated with LPS/IFN-γ (M1) for 12 h. I RT-qPCR analysis of Arg1 and Mrc1 mRNA expression in WT BMDMs treated with various doses of DA and stimulated with IL-4/IL-13 (M2) for 12 h. J Flow cytometry analysis of M2 marker CD206 of WT BMDMs treated with DA (20 μM) and stimulated with IL-4/IL-13 (M2) for 12 h. Representative overlaid flow cytometry histograms showing mean fluorescence intensity (MFI) (left) and quantified data from three independent experiments ( n = 3 per group) (right). K Immunoblot analysis of Arg1 in lysates of WT BMDMs treated with DA (20 μM) and stimulated with M2 for 12 h. L RT-qPCR analysis of Arg1 and Mrc1 mRNA expression in WT and DRD5 −/− BMDMs treated with DA (20 μM) and stimulated with IL-4/IL-13 (M2) for 12 h. M Flow cytometry analysis of M2 marker CD206 of WT and DRD5 −/− BMDMs treated with DA (20 μM) and stimulated with IL-4/IL-13 (M2) for 12 h. Representative overlaid flow cytometry histograms showing mean fluorescence intensity (MFI) (left) and quantified data from three independent experiments ( n = 3 per group) (right). N Immunoblot analysis of Arg1 in lysates of WT and DRD5 −/− BMDMs treated with DA (20 μM) and stimulated with IL-4/IL-13 (M2) for 12 h. Data are pooled from three independent experiments. Error bars show means ± SEM. * p

    Journal: Cell Death & Disease

    Article Title: DA-DRD5 signaling controls colitis by regulating colonic M1/M2 macrophage polarization

    doi: 10.1038/s41419-021-03778-6

    Figure Lengend Snippet: The deficiency of DA-DRD5 signaling inhibits M1 but enhances M2 macrophage polarization in vitro. A RT-qPCR analysis of Inos and Tnf mRNA expression in WT BMDMs treated with various doses of DA and stimulated with LPS/IFN-γ (M1) for 12 h. B ELISA analysis of TNF-α in supernatants from WT BMDMs treated with various doses of DA and stimulated with LPS/IFN-γ (M1) for 12 h. C Flow cytometry analysis of M1 marker CD86 of WT BMDMs treated with DA (20 μM) and stimulated with LPS/IFN-γ (M1) for 12 h. Representative overlaid flow cytometry histograms showing mean fluorescence intensity (MFI) (top) and quantified data from three independent experiments ( n = 3 per group) (bottom). D Immunoblot analysis of Inos in lysates of WT BMDMs treated with DA (20 μM) and stimulated with LPS/IFN-γ (M1) for 12 h. E RT-qPCR analysis of Inos and Tnf mRNA expression in WT and DRD5 −/− BMDMs treated with DA (20 μM) and stimulated with LPS/IFN-γ (M1) for 12 h. F ELISA analysis of TNF-α in supernatants from WT and DRD5 −/− BMDMs treated with DA (20 μM) and stimulated with LPS/IFN-γ (M1) for 12 h. G Flow cytometry analysis of M1 marker CD86 of WT and DRD5 −/− BMDMs treated with DA (20μM) and stimulated with LPS/IFN-γ (M1) for 12 h. Representative overlaid flow cytometry histograms showing mean fluorescence intensity (MFI) (left) and quantified data from three independent experiments ( n = 3 per group) (right). H Immunoblot analysis of Inos in lysates of WT and DRD5 −/− BMDMs treated with DA (20 μM) and stimulated with LPS/IFN-γ (M1) for 12 h. I RT-qPCR analysis of Arg1 and Mrc1 mRNA expression in WT BMDMs treated with various doses of DA and stimulated with IL-4/IL-13 (M2) for 12 h. J Flow cytometry analysis of M2 marker CD206 of WT BMDMs treated with DA (20 μM) and stimulated with IL-4/IL-13 (M2) for 12 h. Representative overlaid flow cytometry histograms showing mean fluorescence intensity (MFI) (left) and quantified data from three independent experiments ( n = 3 per group) (right). K Immunoblot analysis of Arg1 in lysates of WT BMDMs treated with DA (20 μM) and stimulated with M2 for 12 h. L RT-qPCR analysis of Arg1 and Mrc1 mRNA expression in WT and DRD5 −/− BMDMs treated with DA (20 μM) and stimulated with IL-4/IL-13 (M2) for 12 h. M Flow cytometry analysis of M2 marker CD206 of WT and DRD5 −/− BMDMs treated with DA (20 μM) and stimulated with IL-4/IL-13 (M2) for 12 h. Representative overlaid flow cytometry histograms showing mean fluorescence intensity (MFI) (left) and quantified data from three independent experiments ( n = 3 per group) (right). N Immunoblot analysis of Arg1 in lysates of WT and DRD5 −/− BMDMs treated with DA (20 μM) and stimulated with IL-4/IL-13 (M2) for 12 h. Data are pooled from three independent experiments. Error bars show means ± SEM. * p

    Article Snippet: Anti-PKA C-α (55388-1-AP) and anti-Drd5 (20310-1-AP/ADR-005) antibodies were from Proteintech/Alomone.

    Techniques: In Vitro, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Marker, Fluorescence

    GABAergic estrogen-receptive neurons (GERNs) localization, mRNA expression by ISH and somatic receptor expression by IHC. a  Serial coronal sections showing the ERα immunolabelling at the Bregma rostro-caudal coordinates (numbers in mm under the photomicrographs). Boxed area in A 6  at higher magnification showing the exclusive nuclear labeling pattern. The bold numbered levels (A 1  and A 8 ), were chosen to show that no positive labeling was found in either rostral or caudal directions. A 9 : a horizontal view of the distribution of estrogen-receptive cells was symbolized by the red oval. A 10 : sagittal view of rat brain atlas, modified from Paxinos    Watson  86 , at lat. 0.90 mm, where lateral habenula (LHb) is symbolized with a gray shade and A9 plane was symbolized with a horizontal line.  b  In situ hybridization using multiple RNAscope methods. B 1 : multiplex fluorescence method, Esr1, gene that encodes ERα (red punctuated labeling) co-expressed with Slc32a1, gene that encodes VGAT (green punctuated labeling); arrows indicate the double-labeled cells; B 2 : duplex method, Esr1 (red punctuated labeling) co-localization with Slc17a6, gene that encodes VGLUT2 (green punctuated labeling); arrows indicate the double-labeled cells; B 3 : with duplex method, Slc32a1 encoding VGAT (red punctuated labeling) shows complete co-localization with Slc17a6 encoding VGLUT2 (green punctuated labeling); arrows indicate the double-labeled cells. Inset of B 3 , Slc32a1 expression in a sexually active (SA) rat LHb, Br. −3.72 mm (brown labeling, single chromogenic-Brown method-RNAscope). *Note the similarity with ERα expression in A 6 .  c  Indirect immunohistochemistry showing the GABAergic nature of the ERα+ neurons (red) in a SA rat brain. The GABA antibody (green, Sigma, A0310) produced characteristic surface labeling (C 1 , C 2 : the strip-like image was produced by Vibratome slicing leaving the brain section with an uneven surface).  d  ERα+ neurons co-express receptor/receptor subtypes for vasopressin, orexin, dopamine, and serotonin. D 1 : In situ hybridization using RNAscope-multiplex method targeting Esr1 (red dots), Slc32a1 (white dots) and Hcrtr2, gene encodes the orexin receptor 2 (green dots). D 2 , D 3 : Indirect immunofluorescence reactions, showing that the ERα-IR cells co-expressed vasopressin receptor V1a (inset showing the V1a antibody labeling pattern in temporal hippocampus CA2 cells body layer. See also SI Fig.   6  for more information about this antibody), dopamine receptor D5R (also called D1Rb) and serotonin receptor 5-HTr2a, respectively. Scale bars: A 5  and  b : 500 µm and rest: 10 µm

    Journal: Translational Psychiatry

    Article Title: A GABAergic cell type in the lateral habenula links hypothalamic homeostatic and midbrain motivation circuits with sex steroid signaling

    doi: 10.1038/s41398-018-0099-5

    Figure Lengend Snippet: GABAergic estrogen-receptive neurons (GERNs) localization, mRNA expression by ISH and somatic receptor expression by IHC. a Serial coronal sections showing the ERα immunolabelling at the Bregma rostro-caudal coordinates (numbers in mm under the photomicrographs). Boxed area in A 6 at higher magnification showing the exclusive nuclear labeling pattern. The bold numbered levels (A 1 and A 8 ), were chosen to show that no positive labeling was found in either rostral or caudal directions. A 9 : a horizontal view of the distribution of estrogen-receptive cells was symbolized by the red oval. A 10 : sagittal view of rat brain atlas, modified from Paxinos Watson 86 , at lat. 0.90 mm, where lateral habenula (LHb) is symbolized with a gray shade and A9 plane was symbolized with a horizontal line. b In situ hybridization using multiple RNAscope methods. B 1 : multiplex fluorescence method, Esr1, gene that encodes ERα (red punctuated labeling) co-expressed with Slc32a1, gene that encodes VGAT (green punctuated labeling); arrows indicate the double-labeled cells; B 2 : duplex method, Esr1 (red punctuated labeling) co-localization with Slc17a6, gene that encodes VGLUT2 (green punctuated labeling); arrows indicate the double-labeled cells; B 3 : with duplex method, Slc32a1 encoding VGAT (red punctuated labeling) shows complete co-localization with Slc17a6 encoding VGLUT2 (green punctuated labeling); arrows indicate the double-labeled cells. Inset of B 3 , Slc32a1 expression in a sexually active (SA) rat LHb, Br. −3.72 mm (brown labeling, single chromogenic-Brown method-RNAscope). *Note the similarity with ERα expression in A 6 . c Indirect immunohistochemistry showing the GABAergic nature of the ERα+ neurons (red) in a SA rat brain. The GABA antibody (green, Sigma, A0310) produced characteristic surface labeling (C 1 , C 2 : the strip-like image was produced by Vibratome slicing leaving the brain section with an uneven surface). d ERα+ neurons co-express receptor/receptor subtypes for vasopressin, orexin, dopamine, and serotonin. D 1 : In situ hybridization using RNAscope-multiplex method targeting Esr1 (red dots), Slc32a1 (white dots) and Hcrtr2, gene encodes the orexin receptor 2 (green dots). D 2 , D 3 : Indirect immunofluorescence reactions, showing that the ERα-IR cells co-expressed vasopressin receptor V1a (inset showing the V1a antibody labeling pattern in temporal hippocampus CA2 cells body layer. See also SI Fig. 6 for more information about this antibody), dopamine receptor D5R (also called D1Rb) and serotonin receptor 5-HTr2a, respectively. Scale bars: A 5 and b : 500 µm and rest: 10 µm

    Article Snippet: Buijs , 1:2000), tyrosine hydroxylase (sheep anti-TH, EMD Millipore Corporation, MA, AB-1542, 1:4000), serotonin transporter (goat anti-SerT, Santa Cruz Biotechnology, CA, SC-1458, 1:2000), hypocretin/orexin (rabbit anti-OR, gift from A. van del Pol ), vesicular glutamate transporter 2 (guinea pig anti-VGluT2, Frontier Institute, Co., Japan, gp-AF240-1, 1:1000), vesicular inhibitory amino acid transporter (rabbit anti-VGAT/VIAAT, provided by L. E. Eiden , 1:1000), GABA (mouse anti-GABA, Sigma-Aldrich Co. MO, A0310, 1:1000), glutamic acid decarboxylase 65 kDa isoform (mouse anti-GAD-65, EMD Millipore Co. MA, MAB351, 1:2000), glutamic acid decarboxylase 67 kDa isoform (mouse anti-GAD-67, EMD Millipore Co. MA, MAB5406, 1:2000), parvalbumin (mouse anti-PV, Swant, Switzerland, Cat. 235, 1:5000), P450 Aromatase (rabbit anti-ARO, provided by L. M. García-Segura , 1:2000), P450 Aromatase (rabbit anti-ARO, Abcam, Cambrdge, UK, AB18995, 1:2000), P450 Aromatase (mouse anti-ARO, Acris, SM2222P, 1:200), estrogen receptor-alpha (rabbit anti-ERα, Santa Cruz, CA, SC542, 1:2000), androgen receptor (rabbit anti-AR, Santa Cruz, CA, SC816, 1:2000), dopamine receptor 5 (rabbit anti-D5R, also called D1Rb, Alomone, Israel, 1:1000), serotonin receptor (mouse anti-5-HTR2a, BD pharmingen, Cat. 556326, 1:200), vasopressin receptor 1a (rabbit anti-V1a, provided by K. Mutig and T. Giesecke, see and SI Fig. for details), and green fluorescent protein (mouse anti-GFP, Abcam, Cambridge, UK, Ab291-50, 1:500).

    Techniques: Expressing, In Situ Hybridization, Immunohistochemistry, Labeling, Modification, Multiplex Assay, Fluorescence, Produced, Stripping Membranes, Immunofluorescence, Antibody Labeling

    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.

    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:

    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 ††† .

    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:

    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).

    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

    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.

    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