rabbit anti mc4r  (Alomone Labs)


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    Name:
    Anti MC4 Receptor extracellular Antibody
    Description:
    Anti MC4 Receptor extracellular Antibody AMR 024 is a highly specific antibody directed against an epitope of the human melanocortin receptor 4 The antibody can be used in western blot immunohistochemistry and immunocytochemistry applications It recognizes an extracellular epitope and can potentially recognize the receptor in living cells It has been designed to recognize MC4R from human mouse and rat samples
    Catalog Number:
    AMR-024
    Price:
    397.0
    Category:
    Primary Antibody
    Applications:
    Immunocytochemistry, Immunofluorescence, Immunohistochemistry, Live Cell Imaging, Western Blot
    Purity:
    Affinity purified on immobilized antigen.
    Immunogen:
    Synthetic peptide
    Size:
    25 mcl
    Antibody Type:
    Polyclonal Primary Antibodies
    Format:
    Lyophilized Powder
    Host:
    Rabbit
    Isotype:
    Rabbit IgG
    Buy from Supplier


    Structured Review

    Alomone Labs rabbit anti mc4r
    Anti MC4 Receptor extracellular Antibody
    Anti MC4 Receptor extracellular Antibody AMR 024 is a highly specific antibody directed against an epitope of the human melanocortin receptor 4 The antibody can be used in western blot immunohistochemistry and immunocytochemistry applications It recognizes an extracellular epitope and can potentially recognize the receptor in living cells It has been designed to recognize MC4R from human mouse and rat samples
    https://www.bioz.com/result/rabbit anti mc4r/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti mc4r - by Bioz Stars, 2021-09
    93/100 stars

    Images

    1) Product Images from "Activation of the ARCPOMC→MeA Projection Reduces Food Intake"

    Article Title: Activation of the ARCPOMC→MeA Projection Reduces Food Intake

    Journal: Frontiers in Neural Circuits

    doi: 10.3389/fncir.2020.595783

    Activating the ARC POMC →MeA projection leads to an acute reduction in food intake. (A) Schematic drawing of our experimental configurations. A mono fiber-optic cannula was implanted into the MeA of the POMC-Cre; ChR2-tdTomato mice (top panel). The bottom panel shows the implantation site. tdTomato-positive fibers were observed in the MeA of POMC-Cre; ChR2-tdTomato mice. Scale bar: 200 μm. (B) Images of fluorescence confocal microscopy showing that tdTomato-positive fibers were positive for α-MSH in the MeA (white arrowheads). Scale bar: 10 μm. (C) Images of fluorescence confocal microscopy showing that MC4R-positive cells receive POMC input from the ARC. GFP-positive fibers and axonal terminals made synaptic contacts with MC4R-positive cells (white arrowheads). Scale bar: 10 μm. (D) Pooled data from nine male and seven female mice. Twenty hertz stimulation of the ARC POMC →MeA pathway significantly reduced liquid food intake (males, 1.0 ± 0.08 ml vs. 0.6 ± 0.04 ml, ** p
    Figure Legend Snippet: Activating the ARC POMC →MeA projection leads to an acute reduction in food intake. (A) Schematic drawing of our experimental configurations. A mono fiber-optic cannula was implanted into the MeA of the POMC-Cre; ChR2-tdTomato mice (top panel). The bottom panel shows the implantation site. tdTomato-positive fibers were observed in the MeA of POMC-Cre; ChR2-tdTomato mice. Scale bar: 200 μm. (B) Images of fluorescence confocal microscopy showing that tdTomato-positive fibers were positive for α-MSH in the MeA (white arrowheads). Scale bar: 10 μm. (C) Images of fluorescence confocal microscopy showing that MC4R-positive cells receive POMC input from the ARC. GFP-positive fibers and axonal terminals made synaptic contacts with MC4R-positive cells (white arrowheads). Scale bar: 10 μm. (D) Pooled data from nine male and seven female mice. Twenty hertz stimulation of the ARC POMC →MeA pathway significantly reduced liquid food intake (males, 1.0 ± 0.08 ml vs. 0.6 ± 0.04 ml, ** p

    Techniques Used: Microelectrode Array, Mouse Assay, Fluorescence, Confocal Microscopy

    2) Product Images from "Melanocortin 4 Receptor and Dopamine D2 Receptor Expression in Brain Areas Involved in Food Intake"

    Article Title: Melanocortin 4 Receptor and Dopamine D2 Receptor Expression in Brain Areas Involved in Food Intake

    Journal: Endocrinology and Metabolism

    doi: 10.3803/EnM.2015.30.4.576

    Expression of dopamine D2 receptor (D2R) and melanocortin 4 receptor (MC4R) in the striatal area, amygdale, and ventral tegmental area (VTA). Fixed brain slices (40-µm thickness) of D2R-enhanced green fluorescent protein (EGFP) mice were prepared, and immunofluorescence staining for MC4R was performed. Representative immunofluorescence images show D2R-MC4R coexpression in (A) the bed nucleus of the stria terminalis (BNST), (B) central amygdala (CeA), and (C) VTA. The left panel shows a low magnification image (scale bar, 200 µm) and the inset shows a high magnification image (scale bar, 20 µm) for D2R (green) and MC4R (red) stained cells, and D2R-MC4R-coexpressing cells (merge). Arrows indicate D2R-MC4R-coexpressing cells. (D) The graphs show the quantification of D2R- and MC4R-positive cells and D2R-MC4R coexpressed cells in each brain region. Relative percentages of D2R- and MC4R-positive cells were normalized with number of total cells counted. Relative ratio of D2R-MC4R coexpressed cells was shown as a percentage normalized with number of D2R-positive cells. Quantifications were performed in hemisphere of serial sections of each region, on four slices per mouse (2,776 to 3,468 cells) for BNST, six slices per mouse (2,229 to 2,844 cells) for CeA, and five slices per mouse (1,085 to 1,803 cells) for VTA area. The data for each animal are expressed as the average from three mice. CPu, caudate putamen; AC, anterior commissure; NAc, nucleus accumbens; BLA, basolateral amygdala; SNC, substantianigra compact part; SNR, substantianigra reticular part.
    Figure Legend Snippet: Expression of dopamine D2 receptor (D2R) and melanocortin 4 receptor (MC4R) in the striatal area, amygdale, and ventral tegmental area (VTA). Fixed brain slices (40-µm thickness) of D2R-enhanced green fluorescent protein (EGFP) mice were prepared, and immunofluorescence staining for MC4R was performed. Representative immunofluorescence images show D2R-MC4R coexpression in (A) the bed nucleus of the stria terminalis (BNST), (B) central amygdala (CeA), and (C) VTA. The left panel shows a low magnification image (scale bar, 200 µm) and the inset shows a high magnification image (scale bar, 20 µm) for D2R (green) and MC4R (red) stained cells, and D2R-MC4R-coexpressing cells (merge). Arrows indicate D2R-MC4R-coexpressing cells. (D) The graphs show the quantification of D2R- and MC4R-positive cells and D2R-MC4R coexpressed cells in each brain region. Relative percentages of D2R- and MC4R-positive cells were normalized with number of total cells counted. Relative ratio of D2R-MC4R coexpressed cells was shown as a percentage normalized with number of D2R-positive cells. Quantifications were performed in hemisphere of serial sections of each region, on four slices per mouse (2,776 to 3,468 cells) for BNST, six slices per mouse (2,229 to 2,844 cells) for CeA, and five slices per mouse (1,085 to 1,803 cells) for VTA area. The data for each animal are expressed as the average from three mice. CPu, caudate putamen; AC, anterior commissure; NAc, nucleus accumbens; BLA, basolateral amygdala; SNC, substantianigra compact part; SNR, substantianigra reticular part.

    Techniques Used: Expressing, Mouse Assay, Immunofluorescence, Staining

    Expression of dopamine D2 receptor (D2R) and melanocortin 4 receptor (MC4R) in the hypothalamus. D2R-enhanced green fluorescent protein (EGFP) mice were sacrificed, and brain samples were prepared as 40-µm-thick coronal cryosections. Brain sections were then subjected to immunofluorescence staining with anti-MC4R antibody. Immunofluorescence revealed D2R and MC4R coexpression in hypothalamic regions, including (A) the paraventricular nucleus (PVN), (B) arcuate nucleus (ARC), and (C) lateral hypothalamus (LH). The left panel shows a low magnification image (scale bar, 200 µm) and the inset shows a high magnification image (scale bar, 20 µm) for D2R (green) and MC4R (red) stained cells, and D2R-MC4R-coexpressing cells (merge). Arrows indicate D2R-MC4R-coexpressing cells. (D) Relative quantification of D2R- and MC4R-positive cells and D2R-MC4R coexpressed cells in each brain region. Relative percentages of D2R- and MC4R-positive cells were normalized with number of total cells counted. D2R-MC4R coexpression was shown as a percentage normalized with number of D2R-positive cells. Quantifications were performed with hemisphere of coronal slices of each brain region, on six slices per mouse (1,730 to 2,094 cells) for ARC and on six slices per mouse (2,743 to 3,664 cells) for LH area. The data for each animal are expressed as the average from three mice. DMH, dorsomedial hypothalamus; VMH, ventromedial hypothalamus; 3V, third ventricle; CPB, cerebral peduncle basal part; PeF, perifornical nucleus.
    Figure Legend Snippet: Expression of dopamine D2 receptor (D2R) and melanocortin 4 receptor (MC4R) in the hypothalamus. D2R-enhanced green fluorescent protein (EGFP) mice were sacrificed, and brain samples were prepared as 40-µm-thick coronal cryosections. Brain sections were then subjected to immunofluorescence staining with anti-MC4R antibody. Immunofluorescence revealed D2R and MC4R coexpression in hypothalamic regions, including (A) the paraventricular nucleus (PVN), (B) arcuate nucleus (ARC), and (C) lateral hypothalamus (LH). The left panel shows a low magnification image (scale bar, 200 µm) and the inset shows a high magnification image (scale bar, 20 µm) for D2R (green) and MC4R (red) stained cells, and D2R-MC4R-coexpressing cells (merge). Arrows indicate D2R-MC4R-coexpressing cells. (D) Relative quantification of D2R- and MC4R-positive cells and D2R-MC4R coexpressed cells in each brain region. Relative percentages of D2R- and MC4R-positive cells were normalized with number of total cells counted. D2R-MC4R coexpression was shown as a percentage normalized with number of D2R-positive cells. Quantifications were performed with hemisphere of coronal slices of each brain region, on six slices per mouse (1,730 to 2,094 cells) for ARC and on six slices per mouse (2,743 to 3,664 cells) for LH area. The data for each animal are expressed as the average from three mice. DMH, dorsomedial hypothalamus; VMH, ventromedial hypothalamus; 3V, third ventricle; CPB, cerebral peduncle basal part; PeF, perifornical nucleus.

    Techniques Used: Expressing, Mouse Assay, Immunofluorescence, Staining

    3) Product Images from "BMP Receptor 1A Regulates Development of Hypothalamic Circuits Critical for Feeding Behavior"

    Article Title: BMP Receptor 1A Regulates Development of Hypothalamic Circuits Critical for Feeding Behavior

    Journal: The Journal of neuroscience : the official journal of the Society for Neuroscience

    doi: 10.1523/JNEUROSCI.2484-12.2012

    BMP signaling is necessary for neurite outgrowth of the ARH neurons (A–B″) AgRP (red) and TH (green) expression in ARH target nuclei DMH (A-A″) and PVH (B-B″) in P21 control (A′, B′) and R1a KO (A″, B″) hypothalamus. Greatly reduced AgRP + and TH + fibers are detected in both target nuclei. (C) α-MSH (red) and MC4R (green) expression in PVH of P21 control (C′) and R1a KO (C″) hypothalamus. A reduction of α-MSH expression is observed in the R1a KO PVH but MC4R expression is comparable between mutant and control mice. (D) Representative micrograph of P7 wild type ARH explant culture treated with vehicle, leptin (100ng/ml) and leptin plus noggin (250ng/ml) after 3 days in vitro . Extensive neurites outgrowth, represented by β-III tubulin immunostaining away from the tissue (black), is visible in leptin treated group but much fewer in the vehicle control and Noggin treated samples. (E) Quantification of extended neurite specific integrated density in ARH explants treated with vehicle control, leptin, and lepin plus noggin. (F) ARH explants from control and R1a KO mutant mice cultured in the presence of leptin confirmed reduced neurite outgrowth in BMP signaling impaired condition. (G) Quantification of integrated density in control and R1a KO mutant ARH explants. n = 6 to 8 per condition. Value are presented as mean±SEM, **, P
    Figure Legend Snippet: BMP signaling is necessary for neurite outgrowth of the ARH neurons (A–B″) AgRP (red) and TH (green) expression in ARH target nuclei DMH (A-A″) and PVH (B-B″) in P21 control (A′, B′) and R1a KO (A″, B″) hypothalamus. Greatly reduced AgRP + and TH + fibers are detected in both target nuclei. (C) α-MSH (red) and MC4R (green) expression in PVH of P21 control (C′) and R1a KO (C″) hypothalamus. A reduction of α-MSH expression is observed in the R1a KO PVH but MC4R expression is comparable between mutant and control mice. (D) Representative micrograph of P7 wild type ARH explant culture treated with vehicle, leptin (100ng/ml) and leptin plus noggin (250ng/ml) after 3 days in vitro . Extensive neurites outgrowth, represented by β-III tubulin immunostaining away from the tissue (black), is visible in leptin treated group but much fewer in the vehicle control and Noggin treated samples. (E) Quantification of extended neurite specific integrated density in ARH explants treated with vehicle control, leptin, and lepin plus noggin. (F) ARH explants from control and R1a KO mutant mice cultured in the presence of leptin confirmed reduced neurite outgrowth in BMP signaling impaired condition. (G) Quantification of integrated density in control and R1a KO mutant ARH explants. n = 6 to 8 per condition. Value are presented as mean±SEM, **, P

    Techniques Used: Expressing, Mutagenesis, Mouse Assay, In Vitro, Immunostaining, Cell Culture

    Related Articles

    Incubation:

    Article Title: ACTH Prevents Deficits in Fear Extinction Associated with Early Life Seizures
    Article Snippet: .. The sections were then incubated with 400 μL of either 1:400 rabbit Anti-Melanocortin Receptor 3 (extracellular) or 1:700 rabbit Anti-Melanocortin Receptor 4 (extracellular) antibodies (Alomone Labs, Jerusalem, Israel) at 4°C overnight (18 h) on a rocker. ..

    Article Title: Melanocortin 4 Receptor and Dopamine D2 Receptor Expression in Brain Areas Involved in Food Intake
    Article Snippet: .. Sections were then incubated with primary rabbit polyclonal anti-MC4R antibody (1:100, catalog no. AMR-024, Alomone Labs, Jerusalem, Israel) in blocking solution overnight at 4℃ with slow shaking. ..

    Article Title: Activation of the ARCPOMC→MeA Projection Reduces Food Intake
    Article Snippet: .. The sections were blocked in 0.1 M PBS buffer containing 0.2 M glycine, 0.1% Triton X-100, 10% normal donkey serum, and 5% bovine serum albumin for 2 h at room temperature and then incubated with mouse anti-GFP (1:200, Invitrogen, cat# A-11120), rabbit anti-POMC (1:1,000, Phoenix pharmaceuticals, cat# H-029-30), mouse anti-ACTH (1:100, Santa Cruz, cat# sc-57021), sheep anti-α-MSH (1:10,000, Millipore, cat# AB5087), rabbit anti-MC4R (1:250, Alomone labs, cat# AMR-024), and rabbit anti-ER-α (1:40,000, Millipore, cat# 06-935) antibodies for 72 h at cold room. ..

    Blocking Assay:

    Article Title: Melanocortin 4 Receptor and Dopamine D2 Receptor Expression in Brain Areas Involved in Food Intake
    Article Snippet: .. Sections were then incubated with primary rabbit polyclonal anti-MC4R antibody (1:100, catalog no. AMR-024, Alomone Labs, Jerusalem, Israel) in blocking solution overnight at 4℃ with slow shaking. ..

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  • 93
    Alomone Labs rabbit anti mc4r
    Activating the ARC POMC →MeA projection leads to an acute reduction in food intake. (A) Schematic drawing of our experimental configurations. A mono fiber-optic cannula was implanted into the MeA of the POMC-Cre; ChR2-tdTomato mice (top panel). The bottom panel shows the implantation site. tdTomato-positive fibers were observed in the MeA of POMC-Cre; ChR2-tdTomato mice. Scale bar: 200 μm. (B) Images of fluorescence confocal microscopy showing that tdTomato-positive fibers were positive for α-MSH in the MeA (white arrowheads). Scale bar: 10 μm. (C) Images of fluorescence confocal microscopy showing that <t>MC4R-positive</t> cells receive POMC input from the ARC. GFP-positive fibers and axonal terminals made synaptic contacts with MC4R-positive cells (white arrowheads). Scale bar: 10 μm. (D) Pooled data from nine male and seven female mice. Twenty hertz stimulation of the ARC POMC →MeA pathway significantly reduced liquid food intake (males, 1.0 ± 0.08 ml vs. 0.6 ± 0.04 ml, ** p
    Rabbit Anti Mc4r, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti mc4r/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti mc4r - by Bioz Stars, 2021-09
    93/100 stars
      Buy from Supplier

    Image Search Results


    Activating the ARC POMC →MeA projection leads to an acute reduction in food intake. (A) Schematic drawing of our experimental configurations. A mono fiber-optic cannula was implanted into the MeA of the POMC-Cre; ChR2-tdTomato mice (top panel). The bottom panel shows the implantation site. tdTomato-positive fibers were observed in the MeA of POMC-Cre; ChR2-tdTomato mice. Scale bar: 200 μm. (B) Images of fluorescence confocal microscopy showing that tdTomato-positive fibers were positive for α-MSH in the MeA (white arrowheads). Scale bar: 10 μm. (C) Images of fluorescence confocal microscopy showing that MC4R-positive cells receive POMC input from the ARC. GFP-positive fibers and axonal terminals made synaptic contacts with MC4R-positive cells (white arrowheads). Scale bar: 10 μm. (D) Pooled data from nine male and seven female mice. Twenty hertz stimulation of the ARC POMC →MeA pathway significantly reduced liquid food intake (males, 1.0 ± 0.08 ml vs. 0.6 ± 0.04 ml, ** p

    Journal: Frontiers in Neural Circuits

    Article Title: Activation of the ARCPOMC→MeA Projection Reduces Food Intake

    doi: 10.3389/fncir.2020.595783

    Figure Lengend Snippet: Activating the ARC POMC →MeA projection leads to an acute reduction in food intake. (A) Schematic drawing of our experimental configurations. A mono fiber-optic cannula was implanted into the MeA of the POMC-Cre; ChR2-tdTomato mice (top panel). The bottom panel shows the implantation site. tdTomato-positive fibers were observed in the MeA of POMC-Cre; ChR2-tdTomato mice. Scale bar: 200 μm. (B) Images of fluorescence confocal microscopy showing that tdTomato-positive fibers were positive for α-MSH in the MeA (white arrowheads). Scale bar: 10 μm. (C) Images of fluorescence confocal microscopy showing that MC4R-positive cells receive POMC input from the ARC. GFP-positive fibers and axonal terminals made synaptic contacts with MC4R-positive cells (white arrowheads). Scale bar: 10 μm. (D) Pooled data from nine male and seven female mice. Twenty hertz stimulation of the ARC POMC →MeA pathway significantly reduced liquid food intake (males, 1.0 ± 0.08 ml vs. 0.6 ± 0.04 ml, ** p

    Article Snippet: The sections were blocked in 0.1 M PBS buffer containing 0.2 M glycine, 0.1% Triton X-100, 10% normal donkey serum, and 5% bovine serum albumin for 2 h at room temperature and then incubated with mouse anti-GFP (1:200, Invitrogen, cat# A-11120), rabbit anti-POMC (1:1,000, Phoenix pharmaceuticals, cat# H-029-30), mouse anti-ACTH (1:100, Santa Cruz, cat# sc-57021), sheep anti-α-MSH (1:10,000, Millipore, cat# AB5087), rabbit anti-MC4R (1:250, Alomone labs, cat# AMR-024), and rabbit anti-ER-α (1:40,000, Millipore, cat# 06-935) antibodies for 72 h at cold room.

    Techniques: Microelectrode Array, Mouse Assay, Fluorescence, Confocal Microscopy

    Expression of dopamine D2 receptor (D2R) and melanocortin 4 receptor (MC4R) in the striatal area, amygdale, and ventral tegmental area (VTA). Fixed brain slices (40-µm thickness) of D2R-enhanced green fluorescent protein (EGFP) mice were prepared, and immunofluorescence staining for MC4R was performed. Representative immunofluorescence images show D2R-MC4R coexpression in (A) the bed nucleus of the stria terminalis (BNST), (B) central amygdala (CeA), and (C) VTA. The left panel shows a low magnification image (scale bar, 200 µm) and the inset shows a high magnification image (scale bar, 20 µm) for D2R (green) and MC4R (red) stained cells, and D2R-MC4R-coexpressing cells (merge). Arrows indicate D2R-MC4R-coexpressing cells. (D) The graphs show the quantification of D2R- and MC4R-positive cells and D2R-MC4R coexpressed cells in each brain region. Relative percentages of D2R- and MC4R-positive cells were normalized with number of total cells counted. Relative ratio of D2R-MC4R coexpressed cells was shown as a percentage normalized with number of D2R-positive cells. Quantifications were performed in hemisphere of serial sections of each region, on four slices per mouse (2,776 to 3,468 cells) for BNST, six slices per mouse (2,229 to 2,844 cells) for CeA, and five slices per mouse (1,085 to 1,803 cells) for VTA area. The data for each animal are expressed as the average from three mice. CPu, caudate putamen; AC, anterior commissure; NAc, nucleus accumbens; BLA, basolateral amygdala; SNC, substantianigra compact part; SNR, substantianigra reticular part.

    Journal: Endocrinology and Metabolism

    Article Title: Melanocortin 4 Receptor and Dopamine D2 Receptor Expression in Brain Areas Involved in Food Intake

    doi: 10.3803/EnM.2015.30.4.576

    Figure Lengend Snippet: Expression of dopamine D2 receptor (D2R) and melanocortin 4 receptor (MC4R) in the striatal area, amygdale, and ventral tegmental area (VTA). Fixed brain slices (40-µm thickness) of D2R-enhanced green fluorescent protein (EGFP) mice were prepared, and immunofluorescence staining for MC4R was performed. Representative immunofluorescence images show D2R-MC4R coexpression in (A) the bed nucleus of the stria terminalis (BNST), (B) central amygdala (CeA), and (C) VTA. The left panel shows a low magnification image (scale bar, 200 µm) and the inset shows a high magnification image (scale bar, 20 µm) for D2R (green) and MC4R (red) stained cells, and D2R-MC4R-coexpressing cells (merge). Arrows indicate D2R-MC4R-coexpressing cells. (D) The graphs show the quantification of D2R- and MC4R-positive cells and D2R-MC4R coexpressed cells in each brain region. Relative percentages of D2R- and MC4R-positive cells were normalized with number of total cells counted. Relative ratio of D2R-MC4R coexpressed cells was shown as a percentage normalized with number of D2R-positive cells. Quantifications were performed in hemisphere of serial sections of each region, on four slices per mouse (2,776 to 3,468 cells) for BNST, six slices per mouse (2,229 to 2,844 cells) for CeA, and five slices per mouse (1,085 to 1,803 cells) for VTA area. The data for each animal are expressed as the average from three mice. CPu, caudate putamen; AC, anterior commissure; NAc, nucleus accumbens; BLA, basolateral amygdala; SNC, substantianigra compact part; SNR, substantianigra reticular part.

    Article Snippet: Sections were then incubated with primary rabbit polyclonal anti-MC4R antibody (1:100, catalog no. AMR-024, Alomone Labs, Jerusalem, Israel) in blocking solution overnight at 4℃ with slow shaking.

    Techniques: Expressing, Mouse Assay, Immunofluorescence, Staining

    Expression of dopamine D2 receptor (D2R) and melanocortin 4 receptor (MC4R) in the hypothalamus. D2R-enhanced green fluorescent protein (EGFP) mice were sacrificed, and brain samples were prepared as 40-µm-thick coronal cryosections. Brain sections were then subjected to immunofluorescence staining with anti-MC4R antibody. Immunofluorescence revealed D2R and MC4R coexpression in hypothalamic regions, including (A) the paraventricular nucleus (PVN), (B) arcuate nucleus (ARC), and (C) lateral hypothalamus (LH). The left panel shows a low magnification image (scale bar, 200 µm) and the inset shows a high magnification image (scale bar, 20 µm) for D2R (green) and MC4R (red) stained cells, and D2R-MC4R-coexpressing cells (merge). Arrows indicate D2R-MC4R-coexpressing cells. (D) Relative quantification of D2R- and MC4R-positive cells and D2R-MC4R coexpressed cells in each brain region. Relative percentages of D2R- and MC4R-positive cells were normalized with number of total cells counted. D2R-MC4R coexpression was shown as a percentage normalized with number of D2R-positive cells. Quantifications were performed with hemisphere of coronal slices of each brain region, on six slices per mouse (1,730 to 2,094 cells) for ARC and on six slices per mouse (2,743 to 3,664 cells) for LH area. The data for each animal are expressed as the average from three mice. DMH, dorsomedial hypothalamus; VMH, ventromedial hypothalamus; 3V, third ventricle; CPB, cerebral peduncle basal part; PeF, perifornical nucleus.

    Journal: Endocrinology and Metabolism

    Article Title: Melanocortin 4 Receptor and Dopamine D2 Receptor Expression in Brain Areas Involved in Food Intake

    doi: 10.3803/EnM.2015.30.4.576

    Figure Lengend Snippet: Expression of dopamine D2 receptor (D2R) and melanocortin 4 receptor (MC4R) in the hypothalamus. D2R-enhanced green fluorescent protein (EGFP) mice were sacrificed, and brain samples were prepared as 40-µm-thick coronal cryosections. Brain sections were then subjected to immunofluorescence staining with anti-MC4R antibody. Immunofluorescence revealed D2R and MC4R coexpression in hypothalamic regions, including (A) the paraventricular nucleus (PVN), (B) arcuate nucleus (ARC), and (C) lateral hypothalamus (LH). The left panel shows a low magnification image (scale bar, 200 µm) and the inset shows a high magnification image (scale bar, 20 µm) for D2R (green) and MC4R (red) stained cells, and D2R-MC4R-coexpressing cells (merge). Arrows indicate D2R-MC4R-coexpressing cells. (D) Relative quantification of D2R- and MC4R-positive cells and D2R-MC4R coexpressed cells in each brain region. Relative percentages of D2R- and MC4R-positive cells were normalized with number of total cells counted. D2R-MC4R coexpression was shown as a percentage normalized with number of D2R-positive cells. Quantifications were performed with hemisphere of coronal slices of each brain region, on six slices per mouse (1,730 to 2,094 cells) for ARC and on six slices per mouse (2,743 to 3,664 cells) for LH area. The data for each animal are expressed as the average from three mice. DMH, dorsomedial hypothalamus; VMH, ventromedial hypothalamus; 3V, third ventricle; CPB, cerebral peduncle basal part; PeF, perifornical nucleus.

    Article Snippet: Sections were then incubated with primary rabbit polyclonal anti-MC4R antibody (1:100, catalog no. AMR-024, Alomone Labs, Jerusalem, Israel) in blocking solution overnight at 4℃ with slow shaking.

    Techniques: Expressing, Mouse Assay, Immunofluorescence, Staining

    BMP signaling is necessary for neurite outgrowth of the ARH neurons (A–B″) AgRP (red) and TH (green) expression in ARH target nuclei DMH (A-A″) and PVH (B-B″) in P21 control (A′, B′) and R1a KO (A″, B″) hypothalamus. Greatly reduced AgRP + and TH + fibers are detected in both target nuclei. (C) α-MSH (red) and MC4R (green) expression in PVH of P21 control (C′) and R1a KO (C″) hypothalamus. A reduction of α-MSH expression is observed in the R1a KO PVH but MC4R expression is comparable between mutant and control mice. (D) Representative micrograph of P7 wild type ARH explant culture treated with vehicle, leptin (100ng/ml) and leptin plus noggin (250ng/ml) after 3 days in vitro . Extensive neurites outgrowth, represented by β-III tubulin immunostaining away from the tissue (black), is visible in leptin treated group but much fewer in the vehicle control and Noggin treated samples. (E) Quantification of extended neurite specific integrated density in ARH explants treated with vehicle control, leptin, and lepin plus noggin. (F) ARH explants from control and R1a KO mutant mice cultured in the presence of leptin confirmed reduced neurite outgrowth in BMP signaling impaired condition. (G) Quantification of integrated density in control and R1a KO mutant ARH explants. n = 6 to 8 per condition. Value are presented as mean±SEM, **, P

    Journal: The Journal of neuroscience : the official journal of the Society for Neuroscience

    Article Title: BMP Receptor 1A Regulates Development of Hypothalamic Circuits Critical for Feeding Behavior

    doi: 10.1523/JNEUROSCI.2484-12.2012

    Figure Lengend Snippet: BMP signaling is necessary for neurite outgrowth of the ARH neurons (A–B″) AgRP (red) and TH (green) expression in ARH target nuclei DMH (A-A″) and PVH (B-B″) in P21 control (A′, B′) and R1a KO (A″, B″) hypothalamus. Greatly reduced AgRP + and TH + fibers are detected in both target nuclei. (C) α-MSH (red) and MC4R (green) expression in PVH of P21 control (C′) and R1a KO (C″) hypothalamus. A reduction of α-MSH expression is observed in the R1a KO PVH but MC4R expression is comparable between mutant and control mice. (D) Representative micrograph of P7 wild type ARH explant culture treated with vehicle, leptin (100ng/ml) and leptin plus noggin (250ng/ml) after 3 days in vitro . Extensive neurites outgrowth, represented by β-III tubulin immunostaining away from the tissue (black), is visible in leptin treated group but much fewer in the vehicle control and Noggin treated samples. (E) Quantification of extended neurite specific integrated density in ARH explants treated with vehicle control, leptin, and lepin plus noggin. (F) ARH explants from control and R1a KO mutant mice cultured in the presence of leptin confirmed reduced neurite outgrowth in BMP signaling impaired condition. (G) Quantification of integrated density in control and R1a KO mutant ARH explants. n = 6 to 8 per condition. Value are presented as mean±SEM, **, P

    Article Snippet: The primary antibodies used were mouse anti NKX2.1 (1:200; Thermo Fisher), rabbit anti-PAX6 (1:2000; Millipore); rabbit anti-OLIG2 (1: 500; Millipore), goat anti-OLIG2 (1:500; R & D), rabbit anti-ASCL1 (1:1000; BD Pharmingen), gunea pig anti-DLX2 (1:3000), rabbit anti-cleaved Caspase 3 (1:1000; Cell Signaling), rabbit anti-TH (1:1000; Pel Freez), mouse anti-TH (1:500; Sigma), rabbit anti-POMC (1:1000; Phoenix Pharmaceuticals), rabbit anti-NPY (1:1000; Phoenix Pharmaceuticals), rabbit anti-AgRP (1:2000; Phoenix Pharmaceuticals), guinea pig anti-AgRP (1:2000; Abnova), rabbit anti-MC4R (1:250; Alomone Labs), Sheep anti-α-MSH (1:10000, Millipore), rabbit anti-β-Galactosidase (1:2000, Covance), chick anti-β-Galactosidase (1:500, Fitzgerald scientific), rabbit anti-BMPR1A (Abgent AP2004a, 1:200), rabbit anti-phospho-SMAD1/5/8 (1:300, Cell Signaling), rabbit anti-phospho-STAT3 (1:1000, Cell Signaling).

    Techniques: Expressing, Mutagenesis, Mouse Assay, In Vitro, Immunostaining, Cell Culture