damgo  (Millipore)


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    Name:
    Muscimol
    Description:
    Muscimol is a psychoactive isoxazole component which is found in Amanita muscaria and related mushrooms It lowers cerebral glucose metabolism and increases the level of 5 hydroxytryptamine in brain Muscimol has hallucinogenic effects It is associated with distorted perceptions nausea and vomiting
    Catalog Number:
    m1523
    Price:
    None
    Applications:
    Muscimol has been used:. to reduce conditioned fear-expression in lateral amygdala (LA). to eliminate cortical cell response in ferrets. to inactivate lateral magnocellular nucleus of the nidopallium (LMAN) in zebra finches
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    Structured Review

    Millipore damgo
    Muscimol
    Muscimol is a psychoactive isoxazole component which is found in Amanita muscaria and related mushrooms It lowers cerebral glucose metabolism and increases the level of 5 hydroxytryptamine in brain Muscimol has hallucinogenic effects It is associated with distorted perceptions nausea and vomiting
    https://www.bioz.com/result/damgo/product/Millipore
    Average 99 stars, based on 61 article reviews
    Price from $9.99 to $1999.99
    damgo - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "Heterologous desensitization of opioid receptors by chemokines inhibits chemotaxis and enhances the perception of pain"

    Article Title: Heterologous desensitization of opioid receptors by chemokines inhibits chemotaxis and enhances the perception of pain

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi: 10.1073/pnas.102327699

    Restoration of cross-desensitization by readministration of CCL5 or CXCL12. ( A ) CCL5 was administered into the PAG at a concentration of 100 ng, followed by a second administration of CCL5 at the designated concentrations at 120 min. ( B ) Alternatively, CXCL12 was administered into the PAG at a concentration of 100 ng, followed by a second administration of CXCL12 at the designated concentrations at 240 min. After an additional 30 min, DAMGO (400 ng; solid symbols) was administered, and the analgesic response was determined. The analgesic activity of CCL5- or CXCL12-treated mice in the absence of DAMGO is also shown (open symbols). Results are presented as the mean ± SD and are representative of four independent experiments.
    Figure Legend Snippet: Restoration of cross-desensitization by readministration of CCL5 or CXCL12. ( A ) CCL5 was administered into the PAG at a concentration of 100 ng, followed by a second administration of CCL5 at the designated concentrations at 120 min. ( B ) Alternatively, CXCL12 was administered into the PAG at a concentration of 100 ng, followed by a second administration of CXCL12 at the designated concentrations at 240 min. After an additional 30 min, DAMGO (400 ng; solid symbols) was administered, and the analgesic response was determined. The analgesic activity of CCL5- or CXCL12-treated mice in the absence of DAMGO is also shown (open symbols). Results are presented as the mean ± SD and are representative of four independent experiments.

    Techniques Used: Concentration Assay, Activity Assay, Mouse Assay

    The effect of CXCL12, CCL5, and CCL2 on the DAMGO-induced analgesic response in the rat PAG. ( A ) Rats were cannulated into the PAG, and SDF-1α was administered at the designated concentrations. After 30 min, DAMGO (400 ng) was administered, and the analgesic response, expressed as the percent maximum possible analgesia, in the cold-water tail-flick assay was determined. ( B ) Administration of designated concentrations of CXCL12, followed 30 min later with saline, did not show any evidence of detectable analgesia or hyperalgesia. Control experiments showed that the administration of saline, followed 30 min later with saline, also failed to show any evidence of analgesia (data not shown). CCL5 ( C ) or CCL2 ( D ) also was administered into the PAG at the designated concentrations. After 30 min, DAMGO (400 ng; solid symbols) or saline (□) was administered, and the analgesic response was determined. Results are presented as the mean ± SD and are representative of four independent experiments.
    Figure Legend Snippet: The effect of CXCL12, CCL5, and CCL2 on the DAMGO-induced analgesic response in the rat PAG. ( A ) Rats were cannulated into the PAG, and SDF-1α was administered at the designated concentrations. After 30 min, DAMGO (400 ng) was administered, and the analgesic response, expressed as the percent maximum possible analgesia, in the cold-water tail-flick assay was determined. ( B ) Administration of designated concentrations of CXCL12, followed 30 min later with saline, did not show any evidence of detectable analgesia or hyperalgesia. Control experiments showed that the administration of saline, followed 30 min later with saline, also failed to show any evidence of analgesia (data not shown). CCL5 ( C ) or CCL2 ( D ) also was administered into the PAG at the designated concentrations. After 30 min, DAMGO (400 ng; solid symbols) or saline (□) was administered, and the analgesic response was determined. Results are presented as the mean ± SD and are representative of four independent experiments.

    Techniques Used: Tail Flick Test

    Phosphorylation of the μ-opioid receptor after DAMGO or CCL5 treatment. HaCaT cells were treated with either DAMGO (1 nM) or CCL5 (100 ng/ml), and membrane extracts were obtained after 30 min in culture. The extracts were immunoprecipitated with a combination of anti-phosphoserine and anti-phosphothreonine by using protein A/G. The immunoprecipitates were subjected to Western blot analysis by using anti-μ-opioid receptor antibody as a probe, and the reaction was developed by chemiluminescence. The data are representative of four independent experiments.
    Figure Legend Snippet: Phosphorylation of the μ-opioid receptor after DAMGO or CCL5 treatment. HaCaT cells were treated with either DAMGO (1 nM) or CCL5 (100 ng/ml), and membrane extracts were obtained after 30 min in culture. The extracts were immunoprecipitated with a combination of anti-phosphoserine and anti-phosphothreonine by using protein A/G. The immunoprecipitates were subjected to Western blot analysis by using anti-μ-opioid receptor antibody as a probe, and the reaction was developed by chemiluminescence. The data are representative of four independent experiments.

    Techniques Used: Immunoprecipitation, Western Blot

    Duration of cross-desensitization induced by CCL5 or CXCL12. ( A and B ) CCL5 was administered into the PAG at the designated concentrations. After 60 min ( A ) or 120 min ( B ), DAMGO (400 ng; ▾) was administered, and the analgesic response was determined. ( C and D ) Alternatively, CXCL12 was administered into the PAG at the designated concentrations, and after 120 min ( C ) or 240 min ( D ), DAMGO (400 ng; ▾) was administered, and the analgesic response was determined. Results are presented as the mean ± SD and are representative of four independent experiments.
    Figure Legend Snippet: Duration of cross-desensitization induced by CCL5 or CXCL12. ( A and B ) CCL5 was administered into the PAG at the designated concentrations. After 60 min ( A ) or 120 min ( B ), DAMGO (400 ng; ▾) was administered, and the analgesic response was determined. ( C and D ) Alternatively, CXCL12 was administered into the PAG at the designated concentrations, and after 120 min ( C ) or 240 min ( D ), DAMGO (400 ng; ▾) was administered, and the analgesic response was determined. Results are presented as the mean ± SD and are representative of four independent experiments.

    Techniques Used:

    ( A ) Cross-desensitization of the μ-opioid response of monocytes after CCL5 pretreatment. Monocytes were either untreated (•) or pretreated with CCL5 (▾; 100 ng/ml) for 60 min, and the response to the μ-opioid agonist DAMGO was determined. The response of CCL5 pretreated cells to CCL5 (50 ng/ml) is also shown. ( B ) Cross-desensitization of the μ-opioid response of the HaCaT keratinocyte cell line after CCL5 (▴; 100 ng/ml), CCL19 (▵; 100 ng/ml), or CXCL12 (♦; 100 ng/ml) pretreatment. Nontreated cells (circles) and CXCL12-pretreated cells exhibit significant ( P
    Figure Legend Snippet: ( A ) Cross-desensitization of the μ-opioid response of monocytes after CCL5 pretreatment. Monocytes were either untreated (•) or pretreated with CCL5 (▾; 100 ng/ml) for 60 min, and the response to the μ-opioid agonist DAMGO was determined. The response of CCL5 pretreated cells to CCL5 (50 ng/ml) is also shown. ( B ) Cross-desensitization of the μ-opioid response of the HaCaT keratinocyte cell line after CCL5 (▴; 100 ng/ml), CCL19 (▵; 100 ng/ml), or CXCL12 (♦; 100 ng/ml) pretreatment. Nontreated cells (circles) and CXCL12-pretreated cells exhibit significant ( P

    Techniques Used:

    2) Product Images from "Antinociceptive Effects of Nociceptin/Orphanin FQ Administered Intrathecally in Monkeys"

    Article Title: Antinociceptive Effects of Nociceptin/Orphanin FQ Administered Intrathecally in Monkeys

    Journal: The journal of pain : official journal of the American Pain Society

    doi: 10.1016/j.jpain.2008.11.006

    Comparison of itch/scratching responses of intrathecally administered N/OFQ, DAMGO, and substance P. Behavioral responses were scored for each 15-minute session after intrathecal administration of test compound, using a single dosing procedure. Each value
    Figure Legend Snippet: Comparison of itch/scratching responses of intrathecally administered N/OFQ, DAMGO, and substance P. Behavioral responses were scored for each 15-minute session after intrathecal administration of test compound, using a single dosing procedure. Each value

    Techniques Used:

    Comparison of warm water tail-withdrawal responses of intrathecally administered N/OFQ, DAMGO, and substance P. Top panels: Tail-withdrawal latency in 46°C water. Bottom panels: Tail-withdrawal latency in 50°C water. Behavioral responses
    Figure Legend Snippet: Comparison of warm water tail-withdrawal responses of intrathecally administered N/OFQ, DAMGO, and substance P. Top panels: Tail-withdrawal latency in 46°C water. Bottom panels: Tail-withdrawal latency in 50°C water. Behavioral responses

    Techniques Used:

    3) Product Images from "Nucleus Accumbens μ-Opioids Regulate Intake of a High-Fat Diet via Activation of a Distributed Brain Network"

    Article Title: Nucleus Accumbens μ-Opioids Regulate Intake of a High-Fat Diet via Activation of a Distributed Brain Network

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.23-07-02882.2003

    Histological analysis of microinfusion sites for both the muscimol and DAMGO injections. A , Nucleus accumbens placements of one representative study. The remaining slides display the placements of all subjects for each respective study. B , Dorsomedial hypothalamus, lateral hypothalamus, dorsal hippocampus ( C ), ventral tegmental area, ( D .
    Figure Legend Snippet: Histological analysis of microinfusion sites for both the muscimol and DAMGO injections. A , Nucleus accumbens placements of one representative study. The remaining slides display the placements of all subjects for each respective study. B , Dorsomedial hypothalamus, lateral hypothalamus, dorsal hippocampus ( C ), ventral tegmental area, ( D .

    Techniques Used:

    Feeding response to accumbens μ-opioid stimulation (250 ng DAMGO) after muscimol inactivation of selected structures. Values represent group means (±SEM). ∗, DAMGO/SAL versus DAMGO/MUSC; +, DAMGO/SAL versus SAL/SAL; #, MUSC/SAL versus SAL/SAL. Level of significance is shown by number of symbols (* p
    Figure Legend Snippet: Feeding response to accumbens μ-opioid stimulation (250 ng DAMGO) after muscimol inactivation of selected structures. Values represent group means (±SEM). ∗, DAMGO/SAL versus DAMGO/MUSC; +, DAMGO/SAL versus SAL/SAL; #, MUSC/SAL versus SAL/SAL. Level of significance is shown by number of symbols (* p

    Techniques Used:

    4) Product Images from "Methylphenidate and ? opioid receptor interactions: A pharmacological target for prevention of stimulant abuse"

    Article Title: Methylphenidate and ? opioid receptor interactions: A pharmacological target for prevention of stimulant abuse

    Journal: Neuropharmacology

    doi: 10.1016/j.neuropharm.2011.04.015

    High doses of MPH upregulate μ opioid receptor (MOPR) activity in the caudateputamen and nucleus accumbens. [ 35 S]GTPγS binding in membrane preparations from the caudate-putamen was increased by the selective MOPR agonist DAMGO in a concentration-dependent manner with an EC 50 of ~1 and 0.1 mM (A). The maximal binding, which represented 1.75-fold of the basal level was reached at 10 mM concentration (arrow), which was the concentration used in the bindings assays (B). Multiple comparisons analysis showed that the cocaine and high dose MPH (7.5 mg/kg) groups showed significant increases in MOPR activity compared to the saline group (p
    Figure Legend Snippet: High doses of MPH upregulate μ opioid receptor (MOPR) activity in the caudateputamen and nucleus accumbens. [ 35 S]GTPγS binding in membrane preparations from the caudate-putamen was increased by the selective MOPR agonist DAMGO in a concentration-dependent manner with an EC 50 of ~1 and 0.1 mM (A). The maximal binding, which represented 1.75-fold of the basal level was reached at 10 mM concentration (arrow), which was the concentration used in the bindings assays (B). Multiple comparisons analysis showed that the cocaine and high dose MPH (7.5 mg/kg) groups showed significant increases in MOPR activity compared to the saline group (p

    Techniques Used: Activity Assay, Binding Assay, Concentration Assay

    5) Product Images from "Nociceptin/orphanin FQ peptide receptor antagonist JTC-801 reverses pain and anxiety symptoms in a rat model of post-traumatic stress disorder"

    Article Title: Nociceptin/orphanin FQ peptide receptor antagonist JTC-801 reverses pain and anxiety symptoms in a rat model of post-traumatic stress disorder

    Journal: British Journal of Pharmacology

    doi: 10.1111/bph.12701

    Effect of JTC-801 on N/OFQ and DAMGO-stimulated [ 35 S]-GTPγS binding in rat brain. Brain membranes (10 µg) from naïve rats ( n = 4) were incubated with and without agonist and JTC-801 to determine the ability of JTC-801 to block N/OFQ- and DAMGO-mediated increases in [ 35 S]-GTPγS binding, and to alter basal binding. Both N/OFQ and DAMGO (10 −9 –10 −5 M) concentration-dependently stimulated [ 35 S]-GTPγS binding in brain membranes. JTC-801 (1 µM) reversed the stimulatory effect of N/OFQ without altering N/OFQ potency. JTC-801 failed to reverse the stimulatory effect of DAMGO or to alter basal [ 35 S]-GTPγS binding. ** P
    Figure Legend Snippet: Effect of JTC-801 on N/OFQ and DAMGO-stimulated [ 35 S]-GTPγS binding in rat brain. Brain membranes (10 µg) from naïve rats ( n = 4) were incubated with and without agonist and JTC-801 to determine the ability of JTC-801 to block N/OFQ- and DAMGO-mediated increases in [ 35 S]-GTPγS binding, and to alter basal binding. Both N/OFQ and DAMGO (10 −9 –10 −5 M) concentration-dependently stimulated [ 35 S]-GTPγS binding in brain membranes. JTC-801 (1 µM) reversed the stimulatory effect of N/OFQ without altering N/OFQ potency. JTC-801 failed to reverse the stimulatory effect of DAMGO or to alter basal [ 35 S]-GTPγS binding. ** P

    Techniques Used: Binding Assay, Incubation, Blocking Assay, Concentration Assay

    6) Product Images from "Agonists at the ?-opioid receptor modify the binding of u-receptor agonists to the u-? receptor hetero-oligomer"

    Article Title: Agonists at the ?-opioid receptor modify the binding of u-receptor agonists to the u-? receptor hetero-oligomer

    Journal: British Journal of Pharmacology

    doi: 10.1111/j.1476-5381.2010.00944.x

    δ-Opioid agonist detection of the µ-ligand binding pocket in cells co-expressing µ- and δ-opioid receptors (µOR; δOR) or expressing only µ- or δ-receptors. Competition of [ 3 H]-DAMGO binding to membranes from HEK 293T cells expressing only µ-receptors or µ- and δ-receptors by (A) UFP-512 or SNC80 (B), and cells expressing µ- and δ-receptors or only δ-receptors by SNC80 (C). Grey and black arrowheads indicate high and low ligand binding affinities in cells expressing µ-receptors only or the µ–δ receptor heteromer respectively. Cells expressed 150–350 fmol·mg −1 of total receptor protein, whereby each receptor was expressed at a density of 150–175 fmol·mg −1 . Results shown are mean ± SEM, and curves are representative of n = 3–6 experiments performed in duplicate.
    Figure Legend Snippet: δ-Opioid agonist detection of the µ-ligand binding pocket in cells co-expressing µ- and δ-opioid receptors (µOR; δOR) or expressing only µ- or δ-receptors. Competition of [ 3 H]-DAMGO binding to membranes from HEK 293T cells expressing only µ-receptors or µ- and δ-receptors by (A) UFP-512 or SNC80 (B), and cells expressing µ- and δ-receptors or only δ-receptors by SNC80 (C). Grey and black arrowheads indicate high and low ligand binding affinities in cells expressing µ-receptors only or the µ–δ receptor heteromer respectively. Cells expressed 150–350 fmol·mg −1 of total receptor protein, whereby each receptor was expressed at a density of 150–175 fmol·mg −1 . Results shown are mean ± SEM, and curves are representative of n = 3–6 experiments performed in duplicate.

    Techniques Used: Ligand Binding Assay, Expressing, Binding Assay

    Agonist-induced internalization of cell surface µ-opioid receptors (µOR) detected by [ 3 H]-DAMGO in intact HEK 293T cells co-expressing µ- and δ-receptors (δOR), and treated with 100 nM naltrindole or 1 µM CTOP (A), or only expressing µ-receptors (B). Cells co-expressing µ- and δ-receptors were pretreated with 100 nM naltrindole or 1 µM CTOP for 30 min at 37°C, rinsed, then treated with 10 µM agonist for 1 h at 37°C (A). Cells expressing only µ-receptors were treated with 10 µM agonist for 1 h at 37°C (B). Data shown represent drug-induced loss of cell surface receptors as a percentage of cell surface receptors in vehicle-treated control cells, and are expressed as mean ± SEM for n = 3–5 experiments performed in triplicate. Nd = not determined. Statistical significance in (A) was determined using an unpaired Student's t -test [* P
    Figure Legend Snippet: Agonist-induced internalization of cell surface µ-opioid receptors (µOR) detected by [ 3 H]-DAMGO in intact HEK 293T cells co-expressing µ- and δ-receptors (δOR), and treated with 100 nM naltrindole or 1 µM CTOP (A), or only expressing µ-receptors (B). Cells co-expressing µ- and δ-receptors were pretreated with 100 nM naltrindole or 1 µM CTOP for 30 min at 37°C, rinsed, then treated with 10 µM agonist for 1 h at 37°C (A). Cells expressing only µ-receptors were treated with 10 µM agonist for 1 h at 37°C (B). Data shown represent drug-induced loss of cell surface receptors as a percentage of cell surface receptors in vehicle-treated control cells, and are expressed as mean ± SEM for n = 3–5 experiments performed in triplicate. Nd = not determined. Statistical significance in (A) was determined using an unpaired Student's t -test [* P

    Techniques Used: Expressing

    7) Product Images from "Src family kinases mediate the inhibition of substance P release in the rat spinal cord by ?-opioid receptors and GABAB receptors, but not ?2 adrenergic receptors"

    Article Title: Src family kinases mediate the inhibition of substance P release in the rat spinal cord by ?-opioid receptors and GABAB receptors, but not ?2 adrenergic receptors

    Journal: The European journal of neuroscience

    doi: 10.1111/j.1460-9568.2010.07335.x

    NK1R internalization induced by 100 Hz dorsal root stimulation of spinal cord slices: SFK inhibitors reversed the inhibition by DAMGO and baclofen
    Figure Legend Snippet: NK1R internalization induced by 100 Hz dorsal root stimulation of spinal cord slices: SFK inhibitors reversed the inhibition by DAMGO and baclofen

    Techniques Used: Inhibition

    SFK inhibitors reverse the inhibition by DAMGO and baclofen of NK1R internalization induced by noxious stimulation in vivo
    Figure Legend Snippet: SFK inhibitors reverse the inhibition by DAMGO and baclofen of NK1R internalization induced by noxious stimulation in vivo

    Techniques Used: Inhibition, In Vivo

    8) Product Images from "Dynamic monitoring of Gi/o-protein-mediated decreases of intracellular cAMP by FRET-based Epac sensors"

    Article Title: Dynamic monitoring of Gi/o-protein-mediated decreases of intracellular cAMP by FRET-based Epac sensors

    Journal: Pflugers Archiv

    doi: 10.1007/s00424-017-1975-1

    Agonist-induced FRET signal changes can be suppressed by selective receptor antagonists. FRET measurements with HEK293 cells endogenously expressing G s -protein-coupled β 2 Rs or over-expressing G i/o -protein-coupled α 2A or μ receptors together with the FRET-based Epac sensor H74 ( a – c ) or H187 ( d , e ). a – e Representative FRET measurements are displayed with time courses of the normalized yellow and cyan fluorescence signals ( left ) and of the normalized FRET signals ( right ). Hatched bars indicate application of the selective antagonists propranolol (1.5 mM, a , d ), yohimbine (1.0 mM, b ), and CTAP (500 nM, c , e ). Gray bars show application of the agonists isoprenaline (200 μM), guanfacine (200 μM), and DAMGO (100 nM). b , c Application of the adenylyl cyclase activator forskolin (1 μM, FSK) in submaximal concentration to increase basal cAMP levels is displayed
    Figure Legend Snippet: Agonist-induced FRET signal changes can be suppressed by selective receptor antagonists. FRET measurements with HEK293 cells endogenously expressing G s -protein-coupled β 2 Rs or over-expressing G i/o -protein-coupled α 2A or μ receptors together with the FRET-based Epac sensor H74 ( a – c ) or H187 ( d , e ). a – e Representative FRET measurements are displayed with time courses of the normalized yellow and cyan fluorescence signals ( left ) and of the normalized FRET signals ( right ). Hatched bars indicate application of the selective antagonists propranolol (1.5 mM, a , d ), yohimbine (1.0 mM, b ), and CTAP (500 nM, c , e ). Gray bars show application of the agonists isoprenaline (200 μM), guanfacine (200 μM), and DAMGO (100 nM). b , c Application of the adenylyl cyclase activator forskolin (1 μM, FSK) in submaximal concentration to increase basal cAMP levels is displayed

    Techniques Used: Expressing, Fluorescence, Concentration Assay

    9) Product Images from "Opioid-induced Down-Regulation of RGS4"

    Article Title: Opioid-induced Down-Regulation of RGS4

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M110.160911

    The proteasome inhibitor (MG132) and the lysosome inhibitor (leupeptin) increased RGS4 protein levels and prevented the reduction by DAMGO and DPDPE. Left panels , representative Western blot of RGS4 protein expression in SH-SY5Y cells are shown. Right
    Figure Legend Snippet: The proteasome inhibitor (MG132) and the lysosome inhibitor (leupeptin) increased RGS4 protein levels and prevented the reduction by DAMGO and DPDPE. Left panels , representative Western blot of RGS4 protein expression in SH-SY5Y cells are shown. Right

    Techniques Used: Western Blot, Expressing

    10) Product Images from "Mu-Opioid Stimulation in Rat Prefrontal Cortex Engages Hypothalamic Orexin/Hypocretin-Containing Neurons, and Reveals Dissociable Roles of Nucleus Accumbens and Hypothalamus in Cortically Driven Feeding"

    Article Title: Mu-Opioid Stimulation in Rat Prefrontal Cortex Engages Hypothalamic Orexin/Hypocretin-Containing Neurons, and Reveals Dissociable Roles of Nucleus Accumbens and Hypothalamus in Cortically Driven Feeding

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.3323-12.2013

    Effects of AMPA receptor blockade in the AcbSh (with CNQX) on intra-vmPFC DAMGO-induced changes in behavior. Left: Food intake effects following infusion of either DAMGO (2.5 μg/0.5 μl) or saline (Sal) into vmPFC and CNQX into the AcbSh
    Figure Legend Snippet: Effects of AMPA receptor blockade in the AcbSh (with CNQX) on intra-vmPFC DAMGO-induced changes in behavior. Left: Food intake effects following infusion of either DAMGO (2.5 μg/0.5 μl) or saline (Sal) into vmPFC and CNQX into the AcbSh

    Techniques Used:

    Intra-Acb shell infusion of CNQX functionally antagonizes intra-vmPFC DAMGO-induced hyperactivity but augments (additively) DAMGO-induced food intake
    Figure Legend Snippet: Intra-Acb shell infusion of CNQX functionally antagonizes intra-vmPFC DAMGO-induced hyperactivity but augments (additively) DAMGO-induced food intake

    Techniques Used:

    11) Product Images from "Selective κ receptor partial agonist HS666 produces potent antinociception without inducing aversion after i.c.v. administration in mice) Selective κ receptor partial agonist HS666 produces potent antinociception without inducing aversion after i.c.v. administration in mice"

    Article Title: Selective κ receptor partial agonist HS666 produces potent antinociception without inducing aversion after i.c.v. administration in mice) Selective κ receptor partial agonist HS666 produces potent antinociception without inducing aversion after i.c.v. administration in mice

    Journal: British Journal of Pharmacology

    doi: 10.1111/bph.13854

    In vitro functional activity of HS665 and HS666. (A) Stimulation of [ 35 S]‐GTPγS binding to the human μ receptor by HS665, HS666 and DAMGO determined in the [ 35 S]‐GTPγS binding assay with membranes from CHO‐hμ receptor cells. The data were normalized to the maximum stimulation caused by DAMGO (100%) ( n = 3 independent experiments). (B) Stimulation of [ 35 S]‐GTPγS binding to the human κ receptor by HS665, HS666 and U69,593 determined in the [ 35 S]‐GTPγS binding assay with membranes from CHO‐hκ receptor cells. The data were normalized to the maximum stimulation caused by U69,593 (100%) ( n = 5 independent experiments). (C) Selective κ receptor‐mediated G protein activation by HS665 and HS666. Significant antagonism by nor‐BNI (0.1 μM) of the [ 35 S]‐GTPγS binding stimulated by HS665 (1 μM) or HS666 (1 μM) determined in the [ 35 S]‐GTPγS binding assay with membranes from CHO‐hκ receptor cells. The data were normalized to the % stimulation induced by 10 μM U69,593 (100%) ( n = 5 independent experiments). Significantly different from the stimulation with HS665 or HS666 alone, * P
    Figure Legend Snippet: In vitro functional activity of HS665 and HS666. (A) Stimulation of [ 35 S]‐GTPγS binding to the human μ receptor by HS665, HS666 and DAMGO determined in the [ 35 S]‐GTPγS binding assay with membranes from CHO‐hμ receptor cells. The data were normalized to the maximum stimulation caused by DAMGO (100%) ( n = 3 independent experiments). (B) Stimulation of [ 35 S]‐GTPγS binding to the human κ receptor by HS665, HS666 and U69,593 determined in the [ 35 S]‐GTPγS binding assay with membranes from CHO‐hκ receptor cells. The data were normalized to the maximum stimulation caused by U69,593 (100%) ( n = 5 independent experiments). (C) Selective κ receptor‐mediated G protein activation by HS665 and HS666. Significant antagonism by nor‐BNI (0.1 μM) of the [ 35 S]‐GTPγS binding stimulated by HS665 (1 μM) or HS666 (1 μM) determined in the [ 35 S]‐GTPγS binding assay with membranes from CHO‐hκ receptor cells. The data were normalized to the % stimulation induced by 10 μM U69,593 (100%) ( n = 5 independent experiments). Significantly different from the stimulation with HS665 or HS666 alone, * P

    Techniques Used: In Vitro, Functional Assay, Activity Assay, Binding Assay, GTPγS Binding Assay, Activation Assay

    12) Product Images from "In vitro and in vivo pharmacological profile of the 5-benzyl analogue of 14-methoxymetopon, a novel ? opioid analgesic with reduced propensity to alter motor function"

    Article Title: In vitro and in vivo pharmacological profile of the 5-benzyl analogue of 14-methoxymetopon, a novel ? opioid analgesic with reduced propensity to alter motor function

    Journal: European Journal of Pharmaceutical Sciences

    doi: 10.1016/j.ejps.2010.05.018

    Effect of the non-selective opioid antagonist naloxone (NX), and of selective μ (CTAP), δ (NTI) and κ (nor-BNI) opioid receptor antagonists on [ 35 S]GTPγS binding stimulated by compound 1 , 14-MM, 14-OMO and DAMGO in rat brain membranes. Assays were performed in the presence of 1 μM of compound 1 , 14-MM, 14-OMO or DAMGO alone or in the presence of NX (1 μM), CTAP (1 and 10 μM), NTI (10 nM) and nor-BNI (100 nM). Data are shown as % stimulation over basal [ 35 S]GTPγS binding and represent the mean ± SEM of at least three independent experiments, all performed in triplicate. *** p
    Figure Legend Snippet: Effect of the non-selective opioid antagonist naloxone (NX), and of selective μ (CTAP), δ (NTI) and κ (nor-BNI) opioid receptor antagonists on [ 35 S]GTPγS binding stimulated by compound 1 , 14-MM, 14-OMO and DAMGO in rat brain membranes. Assays were performed in the presence of 1 μM of compound 1 , 14-MM, 14-OMO or DAMGO alone or in the presence of NX (1 μM), CTAP (1 and 10 μM), NTI (10 nM) and nor-BNI (100 nM). Data are shown as % stimulation over basal [ 35 S]GTPγS binding and represent the mean ± SEM of at least three independent experiments, all performed in triplicate. *** p

    Techniques Used: Binding Assay

    Concentration-dependent stimulation of [ 35 S]GTPγS binding by compound 1 , and 14-MM, 14-OMO, morphine and DAMGO in rat brain membranes. Data are shown as % stimulation over basal [ 35 S]GTPγS binding and represent the mean ± SEM of at least three independent experiments, all performed in triplicate.
    Figure Legend Snippet: Concentration-dependent stimulation of [ 35 S]GTPγS binding by compound 1 , and 14-MM, 14-OMO, morphine and DAMGO in rat brain membranes. Data are shown as % stimulation over basal [ 35 S]GTPγS binding and represent the mean ± SEM of at least three independent experiments, all performed in triplicate.

    Techniques Used: Concentration Assay, Binding Assay

    13) Product Images from "Mu-Opioid Receptor Coupling to G?o Plays an Important Role in Opioid Antinociception"

    Article Title: Mu-Opioid Receptor Coupling to G?o Plays an Important Role in Opioid Antinociception

    Journal: Neuropsychopharmacology

    doi: 10.1038/npp.2011.91

    DAMGO- and morphine-stimulated G protein activity in spinal cord homogenates from Gα o transgenic mice. [ 35 S]GTPγS (0.1 nM) incorporation stimulated by 10 μM DAMGO or morphine was evaluated in membrane homogenates from spinal cord of wild-type and Gα o +/− mice. Nonspecific binding was evaluated in the presence of unlabeled GTPγS (10 μM). Data are plotted as agonist-stimulated [ 35 S]GTPγS binding, defined as the increase in [ 35 S]GTPγS binding in the presence of agonist over that of basal (measured in the absence of agonist), and represent the mean±SEM ( n =3 performed in quadruplicate). Asterisks indicate a statistical difference vs wild type by Student's paired t -test ( * p
    Figure Legend Snippet: DAMGO- and morphine-stimulated G protein activity in spinal cord homogenates from Gα o transgenic mice. [ 35 S]GTPγS (0.1 nM) incorporation stimulated by 10 μM DAMGO or morphine was evaluated in membrane homogenates from spinal cord of wild-type and Gα o +/− mice. Nonspecific binding was evaluated in the presence of unlabeled GTPγS (10 μM). Data are plotted as agonist-stimulated [ 35 S]GTPγS binding, defined as the increase in [ 35 S]GTPγS binding in the presence of agonist over that of basal (measured in the absence of agonist), and represent the mean±SEM ( n =3 performed in quadruplicate). Asterisks indicate a statistical difference vs wild type by Student's paired t -test ( * p

    Techniques Used: Activity Assay, Transgenic Assay, Mouse Assay, Binding Assay

    14) Product Images from "Endocytosis as a Biological Response in Receptor Pharmacology: Evaluation by Fluorescence Microscopy"

    Article Title: Endocytosis as a Biological Response in Receptor Pharmacology: Evaluation by Fluorescence Microscopy

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0122604

    Time course of the generation of endocytic vesicles containing either DOP-YFP or MOP-YFP receptors transiently expressed in Flp-In T-REx HEK293 cells upon agonist treatment. Time-course curves showing the generation of endocytic vesicles obtained from images of cells expressing DOP-YFP or MOP-YFP receptors upon treatment with DTLET (10 μM) and DAMGO (10 μM) respectively.
    Figure Legend Snippet: Time course of the generation of endocytic vesicles containing either DOP-YFP or MOP-YFP receptors transiently expressed in Flp-In T-REx HEK293 cells upon agonist treatment. Time-course curves showing the generation of endocytic vesicles obtained from images of cells expressing DOP-YFP or MOP-YFP receptors upon treatment with DTLET (10 μM) and DAMGO (10 μM) respectively.

    Techniques Used: Expressing

    15) Product Images from "Multiple Targets of μ-Opioid Receptor-Mediated Presynaptic Inhibition at Primary Afferent Aδ- and C-Fibers"

    Article Title: Multiple Targets of μ-Opioid Receptor-Mediated Presynaptic Inhibition at Primary Afferent Aδ- and C-Fibers

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.4060-10.2011

    MOR agonist DAMGO (10 μ m ) reduced the rate of ionomycin-induced mEPSCs in rat spinal lamina I neurons. A , Representative traces of mEPSC events (control), and mEPSCs, induced by 2 μ m ionomycin, before, during, and after application of
    Figure Legend Snippet: MOR agonist DAMGO (10 μ m ) reduced the rate of ionomycin-induced mEPSCs in rat spinal lamina I neurons. A , Representative traces of mEPSC events (control), and mEPSCs, induced by 2 μ m ionomycin, before, during, and after application of

    Techniques Used:

    16) Product Images from "Dopamine Depletion Reorganizes Projections from the Nucleus Accumbens and Ventral Pallidum That Mediate Opioid-Induced Motor Activity"

    Article Title: Dopamine Depletion Reorganizes Projections from the Nucleus Accumbens and Ventral Pallidum That Mediate Opioid-Induced Motor Activity

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.18-19-08074.1998

    Effect of fluphenazine (9.5 nmol/0.5 μl) microinjections in the nucleus accumbens 5 min before DAMGO microinjections in the nucleus accumbens. The data are shown as mean ± SEM photocell counts (horizontal or rearing activity) or centimeters
    Figure Legend Snippet: Effect of fluphenazine (9.5 nmol/0.5 μl) microinjections in the nucleus accumbens 5 min before DAMGO microinjections in the nucleus accumbens. The data are shown as mean ± SEM photocell counts (horizontal or rearing activity) or centimeters

    Techniques Used: Activity Assay

    Effect of muscimol (0.02 nmol/0.5 μl) in the VP on the motor responses elicited by DAMGO (0.3 nmol/0.5 μl) microinjection into the nucleus accumbens of 6-OHDA-lesioned ( n = 8) or sham-lesioned ( n = 6) rats. Behavior , Left , The data are
    Figure Legend Snippet: Effect of muscimol (0.02 nmol/0.5 μl) in the VP on the motor responses elicited by DAMGO (0.3 nmol/0.5 μl) microinjection into the nucleus accumbens of 6-OHDA-lesioned ( n = 8) or sham-lesioned ( n = 6) rats. Behavior , Left , The data are

    Techniques Used:

    Effect of baclofen (0.1 nmol/0.5 μl) in the VTA on the motor responses elicited by DAMGO (0.3 nmol/0.5 μl) microinjection into the nucleus accumbens. Behavior , Left , The data are shown as mean ± SEM photocell counts (horizontal
    Figure Legend Snippet: Effect of baclofen (0.1 nmol/0.5 μl) in the VTA on the motor responses elicited by DAMGO (0.3 nmol/0.5 μl) microinjection into the nucleus accumbens. Behavior , Left , The data are shown as mean ± SEM photocell counts (horizontal

    Techniques Used:

    Effect of baclofen (0.1 nmol/0.5 μl) in the VTA on the motor responses elicited by DAMGO (0.3 nmol/0.5 μl) microinjection into the VP. Behavior , Left , The data are shown as mean ± SEM photocell counts (horizontal or rearing activity)
    Figure Legend Snippet: Effect of baclofen (0.1 nmol/0.5 μl) in the VTA on the motor responses elicited by DAMGO (0.3 nmol/0.5 μl) microinjection into the VP. Behavior , Left , The data are shown as mean ± SEM photocell counts (horizontal or rearing activity)

    Techniques Used: Activity Assay

    17) Product Images from "Identification and characterization of plant-derived alkaloids, corydine and corydaline, as novel mu opioid receptor agonists"

    Article Title: Identification and characterization of plant-derived alkaloids, corydine and corydaline, as novel mu opioid receptor agonists

    Journal: Scientific Reports

    doi: 10.1038/s41598-020-70493-1

    In vitro activity profiles of corydine ( 1 ) and corydaline ( 2 ) to the human MOR. ( a ) Concentration-dependent inhibition of [ 3 H]DAMGO binding to CHO-hMOR cell membranes was determined in the competitive radioligand binding assay. ( b ) Agonist activities of test compounds to the MOR as stimulation of [ 35 S]GTPγS binding were determined in the [ 35 S]GTPγS binding assay with CHO-hMOR cell membranes. ( c ) β-Arrestin2 recruitment activities of test compounds to the MOR were determined in the PathHunter β-arrestin2 assay. Values are means ± SEM (n = 3–4 independent experiments performed in duplicate).
    Figure Legend Snippet: In vitro activity profiles of corydine ( 1 ) and corydaline ( 2 ) to the human MOR. ( a ) Concentration-dependent inhibition of [ 3 H]DAMGO binding to CHO-hMOR cell membranes was determined in the competitive radioligand binding assay. ( b ) Agonist activities of test compounds to the MOR as stimulation of [ 35 S]GTPγS binding were determined in the [ 35 S]GTPγS binding assay with CHO-hMOR cell membranes. ( c ) β-Arrestin2 recruitment activities of test compounds to the MOR were determined in the PathHunter β-arrestin2 assay. Values are means ± SEM (n = 3–4 independent experiments performed in duplicate).

    Techniques Used: In Vitro, Activity Assay, Concentration Assay, Inhibition, Binding Assay, Radio Ligand Binding Assay, GTPγS Binding Assay

    18) Product Images from "Mechanistic Understanding of Peptide Analogues, DALDA, [Dmt1]DALDA, and KGOP01, Binding to the Mu Opioid Receptor"

    Article Title: Mechanistic Understanding of Peptide Analogues, DALDA, [Dmt1]DALDA, and KGOP01, Binding to the Mu Opioid Receptor

    Journal: Molecules

    doi: 10.3390/molecules25092087

    In vitro activity profiles of DAMGO, DALDA, [Dmt 1 ]DALDA, and KGOP01 to the human MOR (hMOR). ( A ) Binding of tested peptides to the MOR was determined in competitive radioligand binding assays using CHO-hMOR cell membranes. ( B ) Stimulation of [ 35 S]GTPγS binding by tested peptides was determined in the [ 35 S]GTPγS binding assay using CHO-hMOR cell membranes. Values are expressed as the mean ± SEM (n = 3–4 independent experiments).
    Figure Legend Snippet: In vitro activity profiles of DAMGO, DALDA, [Dmt 1 ]DALDA, and KGOP01 to the human MOR (hMOR). ( A ) Binding of tested peptides to the MOR was determined in competitive radioligand binding assays using CHO-hMOR cell membranes. ( B ) Stimulation of [ 35 S]GTPγS binding by tested peptides was determined in the [ 35 S]GTPγS binding assay using CHO-hMOR cell membranes. Values are expressed as the mean ± SEM (n = 3–4 independent experiments).

    Techniques Used: In Vitro, Activity Assay, Binding Assay, GTPγS Binding Assay

    19) Product Images from "Differential modulation of mu-opioid receptor signaling to adenylyl cyclase by RGS proteins 4 or 8 and 7 in permeabilised C6 cells is G? subtype dependent"

    Article Title: Differential modulation of mu-opioid receptor signaling to adenylyl cyclase by RGS proteins 4 or 8 and 7 in permeabilised C6 cells is G? subtype dependent

    Journal: Journal of neurochemistry

    doi: 10.1111/j.1471-4159.2009.06519.x

    Antagonism of RGS8 activity by RGS7-box. Inhibition of forskolin-stimulated AC by DAMGO (100 nM) was measured in the presence of either RGS7-box (5 uM), RGS8 (1 uM) or both (RGS7 + RGS8). Data are expressed as the percentage of the Fsk response and are the mean ± SEM (n=3–4).
    Figure Legend Snippet: Antagonism of RGS8 activity by RGS7-box. Inhibition of forskolin-stimulated AC by DAMGO (100 nM) was measured in the presence of either RGS7-box (5 uM), RGS8 (1 uM) or both (RGS7 + RGS8). Data are expressed as the percentage of the Fsk response and are the mean ± SEM (n=3–4).

    Techniques Used: Activity Assay, Inhibition

    ΔN-RGS4 (RGS4-box) alters mu-opioid inhibition of forskolin-stimulated AC activity. A) Purified ΔN-RGS4 (lacking N-terminal 18 amino-acids) decreased the inhibitory effect of DAMGO (100 nM) in a concentration-dependent manner. Cells were stimulated with 10 µM forskolin (Fsk) ± DAMGO and/or purified RGS protein. cAMP was measured as described. Data are expressed as the percentage of the Fsk response and are the mean ± SEM (n=5–8). B) Increasing concentrations of forskolin (Fsk) were used to stimulate AC activity in the presence (filled circles) or absence (open circles) of a maximal concentration of ΔN-RGS4 (1 µM). Data are expressed as the pmol of cAMP per mg protein*min and are the mean ± SEM from at least four independent experiments (n=4).
    Figure Legend Snippet: ΔN-RGS4 (RGS4-box) alters mu-opioid inhibition of forskolin-stimulated AC activity. A) Purified ΔN-RGS4 (lacking N-terminal 18 amino-acids) decreased the inhibitory effect of DAMGO (100 nM) in a concentration-dependent manner. Cells were stimulated with 10 µM forskolin (Fsk) ± DAMGO and/or purified RGS protein. cAMP was measured as described. Data are expressed as the percentage of the Fsk response and are the mean ± SEM (n=5–8). B) Increasing concentrations of forskolin (Fsk) were used to stimulate AC activity in the presence (filled circles) or absence (open circles) of a maximal concentration of ΔN-RGS4 (1 µM). Data are expressed as the pmol of cAMP per mg protein*min and are the mean ± SEM from at least four independent experiments (n=4).

    Techniques Used: Inhibition, Activity Assay, Purification, Concentration Assay

    Activity of ΔN-RGS4 in cells expressing RGS-insensitive Gαi2. (A) [ 35 S]GTPγS binding was determined in membranes from cells stably expressing either RGS-sensitive (RGS-s) or RGS-insensitive (RGS-i) Gαi2. Overnight treatment with 100 ng/ml pertussis toxin (PTX) was used to block endogenous Gi/o protein activity. Data are derived from four assays, each carried out in duplicate, and are expressed as a percentage of basal binding. Inset: levels of RGS-i and RGS-s Gαi2 expression were verified by SDS-PAGE (20 µg of membranes or 20 ng of Gα i2 standard ( G α i2 )). Proteins were transferred to a nitrocellulose membrane and probed with anti-Gα i2 antibody. Shown is a representative blot from three separate blots. (B) Increasing concentrations of DAMGO were also used to inhibit AC activity stimulated by 10 µM forskolin (Fsk) in RGS-s and RGS-i Gαi2-expressing cells that were permeabilized with digitonin. Again cells were treated overnight with PTX (100 ng/ml) to block coupling to endogenous Gα. Data are the percentage of the Fsk response and are the mean ± SEM from at least three independent experiments. (C) Near maximal concentrations of DAMGO were used to inhibit Fsk-stimulated AC activity in digitonin-permeabilized RGS-s and RGS-i Gαi2-expressing cells (1 µM and 0.1 µM, respectively). Addition of ΔN-RGS4 (1 µM) reduced the inhibitory effect of DAMGO in cells expressing RGS-sensitive Gαi2 but not RGS-insensitive Gαi2. Data are expressed as the percentage of the Fsk response and are the mean ± SEM from at least four independent experiments; *p
    Figure Legend Snippet: Activity of ΔN-RGS4 in cells expressing RGS-insensitive Gαi2. (A) [ 35 S]GTPγS binding was determined in membranes from cells stably expressing either RGS-sensitive (RGS-s) or RGS-insensitive (RGS-i) Gαi2. Overnight treatment with 100 ng/ml pertussis toxin (PTX) was used to block endogenous Gi/o protein activity. Data are derived from four assays, each carried out in duplicate, and are expressed as a percentage of basal binding. Inset: levels of RGS-i and RGS-s Gαi2 expression were verified by SDS-PAGE (20 µg of membranes or 20 ng of Gα i2 standard ( G α i2 )). Proteins were transferred to a nitrocellulose membrane and probed with anti-Gα i2 antibody. Shown is a representative blot from three separate blots. (B) Increasing concentrations of DAMGO were also used to inhibit AC activity stimulated by 10 µM forskolin (Fsk) in RGS-s and RGS-i Gαi2-expressing cells that were permeabilized with digitonin. Again cells were treated overnight with PTX (100 ng/ml) to block coupling to endogenous Gα. Data are the percentage of the Fsk response and are the mean ± SEM from at least three independent experiments. (C) Near maximal concentrations of DAMGO were used to inhibit Fsk-stimulated AC activity in digitonin-permeabilized RGS-s and RGS-i Gαi2-expressing cells (1 µM and 0.1 µM, respectively). Addition of ΔN-RGS4 (1 µM) reduced the inhibitory effect of DAMGO in cells expressing RGS-sensitive Gαi2 but not RGS-insensitive Gαi2. Data are expressed as the percentage of the Fsk response and are the mean ± SEM from at least four independent experiments; *p

    Techniques Used: Activity Assay, Expressing, Binding Assay, Stable Transfection, Blocking Assay, Derivative Assay, SDS Page

    Purified RGS4 and RGS8 negatively regulate opioid coupling to AC. (A) DAMGO inhibition of forskolin-stimulated AC was determined in the presence of maximal concentrations (1 µM) of RGS4 (circles) or RGS8 (squares). Each RGS protein decreased DAMGO potency and efficacy to a similar extent. Data are expressed as the percentage of the Fsk response and are the mean ± SEM from at least four independent experiments. EC 50 values were calculated by non-linear regression (GraphPad Prism). (B) Inhibition of forskolin-stimulated AC by DAMGO (100 nM) was measured in the presence of increasing concentrations of RGS4 (circles), RGS8 (squares), or RGS7-box (triangles). Data are expressed as the percentage of the Fsk response and are the mean ± SEM from four to six independent experiments. (C) Single turnover GTP hydrolysis of Gαo was measured with (filled circles) and without RGS7-box (open circles) to confirm GAP activity of the RGS7-box preparation. Measurements were taken in the presence of 200 nM Gαo ± 125 nM RGS7 with a 3-fold molar excess of γ[ 32 P]GTP. [ 32 P]P i released at each time point was fit to an exponential function: [ 32 P] Pi counts(t) = counts (t=0) + counts (t=30 min) *(1-e −kt ), to calculate the rate constant (k). Fitting constraints included setting counts (t=0) for each curve to the average of the counts (t=0) for the experiment, and setting counts (t=30 min) to the same value for all curves in an experiment. Data are the mean ± SEM of four independent experiments performed in triplicate.
    Figure Legend Snippet: Purified RGS4 and RGS8 negatively regulate opioid coupling to AC. (A) DAMGO inhibition of forskolin-stimulated AC was determined in the presence of maximal concentrations (1 µM) of RGS4 (circles) or RGS8 (squares). Each RGS protein decreased DAMGO potency and efficacy to a similar extent. Data are expressed as the percentage of the Fsk response and are the mean ± SEM from at least four independent experiments. EC 50 values were calculated by non-linear regression (GraphPad Prism). (B) Inhibition of forskolin-stimulated AC by DAMGO (100 nM) was measured in the presence of increasing concentrations of RGS4 (circles), RGS8 (squares), or RGS7-box (triangles). Data are expressed as the percentage of the Fsk response and are the mean ± SEM from four to six independent experiments. (C) Single turnover GTP hydrolysis of Gαo was measured with (filled circles) and without RGS7-box (open circles) to confirm GAP activity of the RGS7-box preparation. Measurements were taken in the presence of 200 nM Gαo ± 125 nM RGS7 with a 3-fold molar excess of γ[ 32 P]GTP. [ 32 P]P i released at each time point was fit to an exponential function: [ 32 P] Pi counts(t) = counts (t=0) + counts (t=30 min) *(1-e −kt ), to calculate the rate constant (k). Fitting constraints included setting counts (t=0) for each curve to the average of the counts (t=0) for the experiment, and setting counts (t=30 min) to the same value for all curves in an experiment. Data are the mean ± SEM of four independent experiments performed in triplicate.

    Techniques Used: Purification, Inhibition, Activity Assay

    20) Product Images from "Dissociation of ?- and ?-opioid inhibition of glutamatergic synaptic transmission in superficial dorsal horn"

    Article Title: Dissociation of ?- and ?-opioid inhibition of glutamatergic synaptic transmission in superficial dorsal horn

    Journal: Molecular Pain

    doi: 10.1186/1744-8069-6-71

    Effect of opioids on basal, and menthol and icilin enhanced miniature EPSCs . (a) Bar chart showing the percentage inhibition of miniature EPSC (mEPSC) rate produced by DAMGO (3 μM), deltorphin-II (Delt, 300 nM) and U69593 (300 nM), expressed as a percentage of the pre-opioid level and averaged across all neurons tested. (b) Bar chart showing the percentage of neurons in which DAMGO, deltorphin-II and U69593 produced a reduction in mEPSC rate of greater than 15%. Data in (a) and (b) are shown for neurons in which opioids were superfused alone, or in the additional presence of menthol (400 μM), or icilin (100 μM). The number in the bars represent (a) the total number of neurons tested and (b) the number of neurons which responded to each opioid.
    Figure Legend Snippet: Effect of opioids on basal, and menthol and icilin enhanced miniature EPSCs . (a) Bar chart showing the percentage inhibition of miniature EPSC (mEPSC) rate produced by DAMGO (3 μM), deltorphin-II (Delt, 300 nM) and U69593 (300 nM), expressed as a percentage of the pre-opioid level and averaged across all neurons tested. (b) Bar chart showing the percentage of neurons in which DAMGO, deltorphin-II and U69593 produced a reduction in mEPSC rate of greater than 15%. Data in (a) and (b) are shown for neurons in which opioids were superfused alone, or in the additional presence of menthol (400 μM), or icilin (100 μM). The number in the bars represent (a) the total number of neurons tested and (b) the number of neurons which responded to each opioid.

    Techniques Used: Inhibition, Produced

    μ-Opioids presynaptically inhibit basal glutamatergic transmission in superficial dorsal horn neurons . (a) Time plot of miniature EPSC (mEPSC) rate during superfusion of deltorphin-II (Delt, 300 nM), DAMGO (3 μM) and naloxone (Nalx, 1 μM). (b) Averaged traces and (c) raw current traces of mEPSCs prior to (Pre), and during deltorphin-II, DAMGO and naloxone. Cumulative probability distribution plots of mEPSC (d) inter-event interval and (e) amplitude for the epochs averaged in (b) (number of events = 635, 640, 327 for control, deltorphin-II and DAMGO, respectively, over 80 s intervals). (a) - (e) are taken from one neuron.
    Figure Legend Snippet: μ-Opioids presynaptically inhibit basal glutamatergic transmission in superficial dorsal horn neurons . (a) Time plot of miniature EPSC (mEPSC) rate during superfusion of deltorphin-II (Delt, 300 nM), DAMGO (3 μM) and naloxone (Nalx, 1 μM). (b) Averaged traces and (c) raw current traces of mEPSCs prior to (Pre), and during deltorphin-II, DAMGO and naloxone. Cumulative probability distribution plots of mEPSC (d) inter-event interval and (e) amplitude for the epochs averaged in (b) (number of events = 635, 640, 327 for control, deltorphin-II and DAMGO, respectively, over 80 s intervals). (a) - (e) are taken from one neuron.

    Techniques Used: Transmission Assay

    Presynaptic μ-opioid inhibition predominates during menthol enhanced glutamatergic transmission . (a) Time plot of miniature EPSC (mEPSC) rate during superfusion of menthol (400 μM), then during addition of deltorphin-II (Delt, 300 nM), ICI-174864 (ICI, 1 μM), U69593 (U69, 300 nM), nor-BNI (300 nM), DAMGO (3 μM) and naloxone (Nalx, 1 μM). (b) Averaged traces and (c) raw current traces of mEPSCs prior to (Pre), and during menthol, then during addition of deltorphin-II, U69593 and DAMGO. Inset in (c) is an expanded part of the trace to show summation of mEPSCs in the presence of menthol. Cumulative probability distribution plots of mEPSC (d) inter-event interval and (e) amplitude for the epochs averaged in (b) (number of events = 648, 1597, 1540, 1259, 506 for control, menthol, deltorphin-II, U69593 and DAMGO over 80, 40, 40, 40 and 80 s intervals, respectively). (a) - (e) are taken from one neuron.
    Figure Legend Snippet: Presynaptic μ-opioid inhibition predominates during menthol enhanced glutamatergic transmission . (a) Time plot of miniature EPSC (mEPSC) rate during superfusion of menthol (400 μM), then during addition of deltorphin-II (Delt, 300 nM), ICI-174864 (ICI, 1 μM), U69593 (U69, 300 nM), nor-BNI (300 nM), DAMGO (3 μM) and naloxone (Nalx, 1 μM). (b) Averaged traces and (c) raw current traces of mEPSCs prior to (Pre), and during menthol, then during addition of deltorphin-II, U69593 and DAMGO. Inset in (c) is an expanded part of the trace to show summation of mEPSCs in the presence of menthol. Cumulative probability distribution plots of mEPSC (d) inter-event interval and (e) amplitude for the epochs averaged in (b) (number of events = 648, 1597, 1540, 1259, 506 for control, menthol, deltorphin-II, U69593 and DAMGO over 80, 40, 40, 40 and 80 s intervals, respectively). (a) - (e) are taken from one neuron.

    Techniques Used: Inhibition, Transmission Assay

    Both μ- and δ-opioids inhibit icilin enhanced glutamatergic transmission . (a) Time plot of miniature EPSC (mEPSC) rate during superfusion of icilin (100 μM), then during addition of DAMGO (3 μM), CTAP (1 μM), deltorphin-II (Delt, 300 nM), ICI-174864 (ICI, 1 μM) and U69593 (U69, 300 nM). (b) Averaged traces and (c) raw current traces of mEPSCs prior to (Pre), and during icilin, then during addition of deltorphin-II, U69593 and DAMGO. Cumulative probability distribution plots of mEPSC (d) inter-event interval and (e) amplitude for the epochs averaged in (b) (number of events = 155, 372, 91, 343, 438 for control, icilin, DAMGO, deltorphin-II and U69593 over 60, 44, 52, 64 and 48 s intervals, respectively). (a) - (e) are taken from one neuron.
    Figure Legend Snippet: Both μ- and δ-opioids inhibit icilin enhanced glutamatergic transmission . (a) Time plot of miniature EPSC (mEPSC) rate during superfusion of icilin (100 μM), then during addition of DAMGO (3 μM), CTAP (1 μM), deltorphin-II (Delt, 300 nM), ICI-174864 (ICI, 1 μM) and U69593 (U69, 300 nM). (b) Averaged traces and (c) raw current traces of mEPSCs prior to (Pre), and during icilin, then during addition of deltorphin-II, U69593 and DAMGO. Cumulative probability distribution plots of mEPSC (d) inter-event interval and (e) amplitude for the epochs averaged in (b) (number of events = 155, 372, 91, 343, 438 for control, icilin, DAMGO, deltorphin-II and U69593 over 60, 44, 52, 64 and 48 s intervals, respectively). (a) - (e) are taken from one neuron.

    Techniques Used: Transmission Assay

    21) Product Images from "Food-Derived Hemorphins Cross Intestinal and Blood–Brain Barriers In Vitro"

    Article Title: Food-Derived Hemorphins Cross Intestinal and Blood–Brain Barriers In Vitro

    Journal: Frontiers in Endocrinology

    doi: 10.3389/fendo.2018.00159

    Determination of intracellular cAMP in forskolin (FK)-stimulated Caco-2 cells after incubation with the five hemorphins. (A) The ability of the hemorphins/opioid peptides to act on the cAMP pathway was evaluated in vitro . Caco-2 cells were incubated with or without increasing concentrations of each hemorphin, DAMGO, a known specific OP agonist and naloxone, a known OP antagonist, in FK-supplemented medium for 15 min at 37°C. Intracellular cAMP levels were determined by ELISA, normalized by the total protein concentration and expressed as a percentage of the reference group FK. Data presented are mean ± SD (average of two assays performed in triplicate; n = 6). *Statistically different from control; * p
    Figure Legend Snippet: Determination of intracellular cAMP in forskolin (FK)-stimulated Caco-2 cells after incubation with the five hemorphins. (A) The ability of the hemorphins/opioid peptides to act on the cAMP pathway was evaluated in vitro . Caco-2 cells were incubated with or without increasing concentrations of each hemorphin, DAMGO, a known specific OP agonist and naloxone, a known OP antagonist, in FK-supplemented medium for 15 min at 37°C. Intracellular cAMP levels were determined by ELISA, normalized by the total protein concentration and expressed as a percentage of the reference group FK. Data presented are mean ± SD (average of two assays performed in triplicate; n = 6). *Statistically different from control; * p

    Techniques Used: Incubation, Activated Clotting Time Assay, In Vitro, Enzyme-linked Immunosorbent Assay, Protein Concentration

    22) Product Images from "Dopamine D4 Receptor Counteracts Morphine-Induced Changes in ? Opioid Receptor Signaling in the Striosomes of the Rat Caudate Putamen"

    Article Title: Dopamine D4 Receptor Counteracts Morphine-Induced Changes in ? Opioid Receptor Signaling in the Striosomes of the Rat Caudate Putamen

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms15011481

    Co-administration of PD168,077 during continuous morphine treatment prevents the increase of [ 3 H]DAMGO binding sites induced by the opioid drug. ( A – D ) Representative autoradiograms from coronal brain sections at the CPu level of rats which received six days of continuous treatment with vehicle ( A ), morphine (20 mg/kg/day) ( B ); PD168,077 (1 mg/kg/day) ( C ) and morphine + PD168,077 ( D ); Abbreviations: Cx, cortex; CPu, caudate putamen; NAc, nucleus accumbens. Scale bar is 2 mm. ( E ) Saturation curves of [ 3 H]DAMGO binding in the striosomes; ( F , G ) Effect of drug treatments on K d and B max values (mean ± SEM; n = 6) of [ 3 H]DAMGO binding in the striosomes and matrix compartment of the rat CPu. * p
    Figure Legend Snippet: Co-administration of PD168,077 during continuous morphine treatment prevents the increase of [ 3 H]DAMGO binding sites induced by the opioid drug. ( A – D ) Representative autoradiograms from coronal brain sections at the CPu level of rats which received six days of continuous treatment with vehicle ( A ), morphine (20 mg/kg/day) ( B ); PD168,077 (1 mg/kg/day) ( C ) and morphine + PD168,077 ( D ); Abbreviations: Cx, cortex; CPu, caudate putamen; NAc, nucleus accumbens. Scale bar is 2 mm. ( E ) Saturation curves of [ 3 H]DAMGO binding in the striosomes; ( F , G ) Effect of drug treatments on K d and B max values (mean ± SEM; n = 6) of [ 3 H]DAMGO binding in the striosomes and matrix compartment of the rat CPu. * p

    Techniques Used: Binding Assay

    D 4 R in vitro activation prevents morphine-induced changes on MOR-dependent [ 35 S]GTPγS binding. ( A – D3 ) Representative autoradiograms of [ 35 S]GTPγS binding in coronal sections of rat brain at the CPu level. Rats were continuously treated with vehicle ( A – A3 ); morphine (20 mg/kg/day) ( B – B3 ); PD168,077 (1 mg/kg/day) ( C – C3 ) and morphine + PD168,077 ( D – D3 ); Basal levels of [ 35 S]GTPγS binding was determined in control sections from the four treatment groups ( A , B , C , D ) and in vitro receptor stimulation was performed with DAMGO (3 μM) ( A1 , B1 , C1 , D1 ); PD168,077 (90 nM) ( A2 , B2 , C2 , D2 ) or DAMGO + PD168,077 ( A3 , B3 , C3 , D3 ). Scale bar is 2 mm; ( E ) Effect of continuous drug treatments on [ 35 S]GTPγS binding in the rat CPu after in vitro agonist stimulation of MOR and/or D 4 R. Data represent mean ± SEM ( n = 6) and are expressed as percentage of basal [ 35 S]GTPγS binding value in vehicle-treated animals (red line). Blue line represents DAMGO-dependent [ 35 S]GTPγS binding in vehicle-treated animals. Differences between groups were set by two-way ANOVA followed by post hoc Bonferroni t test. * p
    Figure Legend Snippet: D 4 R in vitro activation prevents morphine-induced changes on MOR-dependent [ 35 S]GTPγS binding. ( A – D3 ) Representative autoradiograms of [ 35 S]GTPγS binding in coronal sections of rat brain at the CPu level. Rats were continuously treated with vehicle ( A – A3 ); morphine (20 mg/kg/day) ( B – B3 ); PD168,077 (1 mg/kg/day) ( C – C3 ) and morphine + PD168,077 ( D – D3 ); Basal levels of [ 35 S]GTPγS binding was determined in control sections from the four treatment groups ( A , B , C , D ) and in vitro receptor stimulation was performed with DAMGO (3 μM) ( A1 , B1 , C1 , D1 ); PD168,077 (90 nM) ( A2 , B2 , C2 , D2 ) or DAMGO + PD168,077 ( A3 , B3 , C3 , D3 ). Scale bar is 2 mm; ( E ) Effect of continuous drug treatments on [ 35 S]GTPγS binding in the rat CPu after in vitro agonist stimulation of MOR and/or D 4 R. Data represent mean ± SEM ( n = 6) and are expressed as percentage of basal [ 35 S]GTPγS binding value in vehicle-treated animals (red line). Blue line represents DAMGO-dependent [ 35 S]GTPγS binding in vehicle-treated animals. Differences between groups were set by two-way ANOVA followed by post hoc Bonferroni t test. * p

    Techniques Used: In Vitro, Activation Assay, Binding Assay

    23) Product Images from "Dopamine D4 Receptor Counteracts Morphine-Induced Changes in ? Opioid Receptor Signaling in the Striosomes of the Rat Caudate Putamen"

    Article Title: Dopamine D4 Receptor Counteracts Morphine-Induced Changes in ? Opioid Receptor Signaling in the Striosomes of the Rat Caudate Putamen

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms15011481

    Co-administration of PD168,077 during continuous morphine treatment prevents the increase of [ 3 H]DAMGO binding sites induced by the opioid drug. ( A – D ) Representative autoradiograms from coronal brain sections at the CPu level of rats which received six days of continuous treatment with vehicle ( A ), morphine (20 mg/kg/day) ( B ); PD168,077 (1 mg/kg/day) ( C ) and morphine + PD168,077 ( D ); Abbreviations: Cx, cortex; CPu, caudate putamen; NAc, nucleus accumbens. Scale bar is 2 mm. ( E ) Saturation curves of [ 3 H]DAMGO binding in the striosomes; ( F , G ) Effect of drug treatments on K d and B max values (mean ± SEM; n = 6) of [ 3 H]DAMGO binding in the striosomes and matrix compartment of the rat CPu. * p
    Figure Legend Snippet: Co-administration of PD168,077 during continuous morphine treatment prevents the increase of [ 3 H]DAMGO binding sites induced by the opioid drug. ( A – D ) Representative autoradiograms from coronal brain sections at the CPu level of rats which received six days of continuous treatment with vehicle ( A ), morphine (20 mg/kg/day) ( B ); PD168,077 (1 mg/kg/day) ( C ) and morphine + PD168,077 ( D ); Abbreviations: Cx, cortex; CPu, caudate putamen; NAc, nucleus accumbens. Scale bar is 2 mm. ( E ) Saturation curves of [ 3 H]DAMGO binding in the striosomes; ( F , G ) Effect of drug treatments on K d and B max values (mean ± SEM; n = 6) of [ 3 H]DAMGO binding in the striosomes and matrix compartment of the rat CPu. * p

    Techniques Used: Binding Assay

    D 4 R in vitro activation prevents morphine-induced changes on MOR-dependent [ 35 S]GTPγS binding. ( A – D3 ) Representative autoradiograms of [ 35 S]GTPγS binding in coronal sections of rat brain at the CPu level. Rats were continuously treated with vehicle ( A – A3 ); morphine (20 mg/kg/day) ( B – B3 ); PD168,077 (1 mg/kg/day) ( C – C3 ) and morphine + PD168,077 ( D – D3 ); Basal levels of [ 35 S]GTPγS binding was determined in control sections from the four treatment groups ( A , B , C , D ) and in vitro receptor stimulation was performed with DAMGO (3 μM) ( A1 , B1 , C1 , D1 ); PD168,077 (90 nM) ( A2 , B2 , C2 , D2 ) or DAMGO + PD168,077 ( A3 , B3 , C3 , D3 ). Scale bar is 2 mm; ( E ) Effect of continuous drug treatments on [ 35 S]GTPγS binding in the rat CPu after in vitro agonist stimulation of MOR and/or D 4 R. Data represent mean ± SEM ( n = 6) and are expressed as percentage of basal [ 35 S]GTPγS binding value in vehicle-treated animals (red line). Blue line represents DAMGO-dependent [ 35 S]GTPγS binding in vehicle-treated animals. Differences between groups were set by two-way ANOVA followed by post hoc Bonferroni t test. * p
    Figure Legend Snippet: D 4 R in vitro activation prevents morphine-induced changes on MOR-dependent [ 35 S]GTPγS binding. ( A – D3 ) Representative autoradiograms of [ 35 S]GTPγS binding in coronal sections of rat brain at the CPu level. Rats were continuously treated with vehicle ( A – A3 ); morphine (20 mg/kg/day) ( B – B3 ); PD168,077 (1 mg/kg/day) ( C – C3 ) and morphine + PD168,077 ( D – D3 ); Basal levels of [ 35 S]GTPγS binding was determined in control sections from the four treatment groups ( A , B , C , D ) and in vitro receptor stimulation was performed with DAMGO (3 μM) ( A1 , B1 , C1 , D1 ); PD168,077 (90 nM) ( A2 , B2 , C2 , D2 ) or DAMGO + PD168,077 ( A3 , B3 , C3 , D3 ). Scale bar is 2 mm; ( E ) Effect of continuous drug treatments on [ 35 S]GTPγS binding in the rat CPu after in vitro agonist stimulation of MOR and/or D 4 R. Data represent mean ± SEM ( n = 6) and are expressed as percentage of basal [ 35 S]GTPγS binding value in vehicle-treated animals (red line). Blue line represents DAMGO-dependent [ 35 S]GTPγS binding in vehicle-treated animals. Differences between groups were set by two-way ANOVA followed by post hoc Bonferroni t test. * p

    Techniques Used: In Vitro, Activation Assay, Binding Assay

    24) Product Images from "Multiple Receptors Involved in Peripheral α2, μ, and A1 Antinociception, Tolerance, and Withdrawal"

    Article Title: Multiple Receptors Involved in Peripheral α2, μ, and A1 Antinociception, Tolerance, and Withdrawal

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.17-02-00735.1997

    Antisense μ ODN treatment blocks not only μ antinociception but also α 2 antinociception. A 1 antinociception is unaffected. A , Effect of PGE 2 ( E2 ), DAMGO plus PGE 2 ( D+E2 ), μ-antisense ( AS ) ODN 1 μg intrathecally on alternate days × 3 and DAMGO plus PGE 2 [ μ-(AS)x3,D+E2 ], μ-sense ( S ) ODN 1 μg intrathecally on alternate days × 3, and DAMGO plus PGE 2 [ μ-(S)x3,D+E2 ] on mechanical paw-withdrawal threshold. B , Effect of PGE 2 ( E2 ), clonidine plus PGE 2 ( Cl+E2 ), μ-(AS) ODN 1 μg intrathecally on alternate days × 3, and clonidine plus PGE 2 [ μ-(AS)x3,Cl+E2 ], μ-(S) ODN 1 μg intrathecally on alternate days × 3, and clonidine plus PGE 2 [ μ-(S)x3,Cl+E2 ] on mechanical paw-withdrawal threshold. C , Effect of PGE 2 ( E2 ), CPA plus PGE 2 ( CPA+E2 ), μ-(AS) ODN 1 μg intrathecally on alternate days × 3, CPA plus PGE 2 [ μ-(AS)x3,CPA+E2 ], μ-(S) ODN 1 μg intrathecally on alternate days × 3, and CPA plus PGE 2 [ μ-(S)x3,CPA+E2 ] on mechanical paw withdrawal threshold in the rat.
    Figure Legend Snippet: Antisense μ ODN treatment blocks not only μ antinociception but also α 2 antinociception. A 1 antinociception is unaffected. A , Effect of PGE 2 ( E2 ), DAMGO plus PGE 2 ( D+E2 ), μ-antisense ( AS ) ODN 1 μg intrathecally on alternate days × 3 and DAMGO plus PGE 2 [ μ-(AS)x3,D+E2 ], μ-sense ( S ) ODN 1 μg intrathecally on alternate days × 3, and DAMGO plus PGE 2 [ μ-(S)x3,D+E2 ] on mechanical paw-withdrawal threshold. B , Effect of PGE 2 ( E2 ), clonidine plus PGE 2 ( Cl+E2 ), μ-(AS) ODN 1 μg intrathecally on alternate days × 3, and clonidine plus PGE 2 [ μ-(AS)x3,Cl+E2 ], μ-(S) ODN 1 μg intrathecally on alternate days × 3, and clonidine plus PGE 2 [ μ-(S)x3,Cl+E2 ] on mechanical paw-withdrawal threshold. C , Effect of PGE 2 ( E2 ), CPA plus PGE 2 ( CPA+E2 ), μ-(AS) ODN 1 μg intrathecally on alternate days × 3, CPA plus PGE 2 [ μ-(AS)x3,CPA+E2 ], μ-(S) ODN 1 μg intrathecally on alternate days × 3, and CPA plus PGE 2 [ μ-(S)x3,CPA+E2 ] on mechanical paw withdrawal threshold in the rat.

    Techniques Used:

    Multiple receptors are involved in α 2 , μ, and A 1 tolerance and withdrawal. A , Yohimbine withdrawal is blocked not only by clonidine but also by DAMGO and CPA. Effect of clonidine once hourly for 3 hr and at the fourth hour yohimbine ( Clx3,Yo ), clonidine once hourly for 3 hr and at the fourth hour clonidine plus yohimbine ( Clx3,Cl+Yo ), clonidine once hourly for 3 hr and at the fourth hour DAMGO plus yohimbine ( Clx3,D+Yo ), and clonidine once hourly for 3 hr and at the fourth hour CPA plus yohimbine ( Clx3,CPA+Yo ) on mechanical paw withdrawal threshold in the rat. B , Naloxone withdrawal is blocked not only by DAMGO but also by clonidine. Effect of DAMGO once hourly for 3 hr and at the fourth hour naloxone ( Dx3,N ), DAMGO once hourly for 3 hr and at the fourth hour DAMGO plus naloxone ( Dx3,D+N ), DAMGO once hourly for 3 hr and at the fourth hour clonidine plus naloxone ( Dx3,Cl+N ), and DAMGO once hourly for 3 hr and at the fourth hour CPA plus naloxone ( Dx3,CPA+N ) on mechanical paw withdrawal threshold in the rat. C , PACPX withdrawal is blocked not only by CPA but also by clonidine. Effect of CPA once hourly for 3 hr and at the fourth hour PACPX ( CPAx3,PACPX ), CPA once hourly for 3 hr and at the fourth hour CPA plus PACPX ( CPAx3,CPA+PACPX ), CPA once hourly for 3 hr and at the fourth hour clonidine plus PACPX ( CPAx3,Cl+PACPX ), and CPA once hourly for 3 hr and at the fourth hour DAMGO plus naloxone ( CPAx3,D+PACPX ) on mechanical paw withdrawal threshold in the rat.
    Figure Legend Snippet: Multiple receptors are involved in α 2 , μ, and A 1 tolerance and withdrawal. A , Yohimbine withdrawal is blocked not only by clonidine but also by DAMGO and CPA. Effect of clonidine once hourly for 3 hr and at the fourth hour yohimbine ( Clx3,Yo ), clonidine once hourly for 3 hr and at the fourth hour clonidine plus yohimbine ( Clx3,Cl+Yo ), clonidine once hourly for 3 hr and at the fourth hour DAMGO plus yohimbine ( Clx3,D+Yo ), and clonidine once hourly for 3 hr and at the fourth hour CPA plus yohimbine ( Clx3,CPA+Yo ) on mechanical paw withdrawal threshold in the rat. B , Naloxone withdrawal is blocked not only by DAMGO but also by clonidine. Effect of DAMGO once hourly for 3 hr and at the fourth hour naloxone ( Dx3,N ), DAMGO once hourly for 3 hr and at the fourth hour DAMGO plus naloxone ( Dx3,D+N ), DAMGO once hourly for 3 hr and at the fourth hour clonidine plus naloxone ( Dx3,Cl+N ), and DAMGO once hourly for 3 hr and at the fourth hour CPA plus naloxone ( Dx3,CPA+N ) on mechanical paw withdrawal threshold in the rat. C , PACPX withdrawal is blocked not only by CPA but also by clonidine. Effect of CPA once hourly for 3 hr and at the fourth hour PACPX ( CPAx3,PACPX ), CPA once hourly for 3 hr and at the fourth hour CPA plus PACPX ( CPAx3,CPA+PACPX ), CPA once hourly for 3 hr and at the fourth hour clonidine plus PACPX ( CPAx3,Cl+PACPX ), and CPA once hourly for 3 hr and at the fourth hour DAMGO plus naloxone ( CPAx3,D+PACPX ) on mechanical paw withdrawal threshold in the rat.

    Techniques Used:

    A , Yohimbine precipitates withdrawal hyperalgesia in clonidine tolerant paws. Effect of PGE 2 ( E2 ), clonidine plus PGE 2 ( Cl+E2 ), clonidine once hourly for 3 hr ( Clx3 ), clonidine once hourly for 3 hr ( Clx3 ), clonidine once hourly for 3 hr, and at the fourth hour yohimbine ( Clx3, Yo ) on mechanical paw-withdrawal threshold in the rat. B , Bidirectional cross-withdrawal develops among A 1 , α 2 , and μ antinociception. Effect of clonidine once hourly for 3 hr and at the fourth hour naloxone ( Clx3, N ), clonidine once hourly for 3 hr and at the fourth hour PACPX ( Clx3, PACPX ), clonidine once hourly for 3 hr ( Clx3 ), DAMGO once hourly for 3 hr and at the fourth hour yohimbine ( Dx3, Yo ), DAMGO once hourly for 3 hr ( Dx3 ), CPA once hourly for 3 hr ( CPAx3 ), CPA once hourly for 3 hr and at the fourth hour yohimbine ( CPAx3, Yo ), and CPA once hourly for 3 hr ( CPAx3 ) on mechanical paw withdrawal threshold in the rat.
    Figure Legend Snippet: A , Yohimbine precipitates withdrawal hyperalgesia in clonidine tolerant paws. Effect of PGE 2 ( E2 ), clonidine plus PGE 2 ( Cl+E2 ), clonidine once hourly for 3 hr ( Clx3 ), clonidine once hourly for 3 hr ( Clx3 ), clonidine once hourly for 3 hr, and at the fourth hour yohimbine ( Clx3, Yo ) on mechanical paw-withdrawal threshold in the rat. B , Bidirectional cross-withdrawal develops among A 1 , α 2 , and μ antinociception. Effect of clonidine once hourly for 3 hr and at the fourth hour naloxone ( Clx3, N ), clonidine once hourly for 3 hr and at the fourth hour PACPX ( Clx3, PACPX ), clonidine once hourly for 3 hr ( Clx3 ), DAMGO once hourly for 3 hr and at the fourth hour yohimbine ( Dx3, Yo ), DAMGO once hourly for 3 hr ( Dx3 ), CPA once hourly for 3 hr ( CPAx3 ), CPA once hourly for 3 hr and at the fourth hour yohimbine ( CPAx3, Yo ), and CPA once hourly for 3 hr ( CPAx3 ) on mechanical paw withdrawal threshold in the rat.

    Techniques Used:

    Schematic diagram of hypothesized topological/physical arrangement of the three receptors for peripheral antinociception in the cell membrane. μ ( DAMGO ), α 2C ( Clonidine ), and A 1 ( CPA ) agonism all result in peripheral antinociception mediated through a common second messenger pathway, leading to complete symmetrical cross-tolerance and cross-dependence. However, the asymmetrical interactions are proposed to be a result of the central position of the α 2C receptor leading to bidirectional interactions between this receptor and the two other receptors but no interaction between the μ and A 1 receptors.
    Figure Legend Snippet: Schematic diagram of hypothesized topological/physical arrangement of the three receptors for peripheral antinociception in the cell membrane. μ ( DAMGO ), α 2C ( Clonidine ), and A 1 ( CPA ) agonism all result in peripheral antinociception mediated through a common second messenger pathway, leading to complete symmetrical cross-tolerance and cross-dependence. However, the asymmetrical interactions are proposed to be a result of the central position of the α 2C receptor leading to bidirectional interactions between this receptor and the two other receptors but no interaction between the μ and A 1 receptors.

    Techniques Used:

    A , Repeated administration of clonidine produces tolerance to antinociception. Effect of PGE 2 ( E2 ), clonidine plus PGE 2 ( Cl+E2 ), clonidine once hourly for 3 hr ( Clx3 ), clonidine once hourly for 3 hr, and at the fourth hour clonidine plus PGE 2 ( Clx3, Cl+E2 ) on mechanical paw withdrawal threshold in the rat. B , Bidirectional cross-tolerance develops among A 1 , α 2 , and μ antinociception. Effect of clonidine plus PGE 2 ( Cl+E2 ), DAMGO once hourly for 3 hr ( Dx3 ), DAMGO once hourly for 3 hr and at the fourth hour clonidine plus PGE 2 ( Dx3,Cl+E2 ), CPA once hourly for 3 hr and at the fourth hour clonidine plus PGE 2 ( CPAx3, Cl+E2 ), CPA once hourly for 3 hr ( CPAx3 ), clonidine once hourly for 3 hr and at the fourth hour DAMGO plus PGE 2 ( Clx3, D+E2 ), and clonidine once hourly for 3 hr and at the fourth hour CPA plus PGE 2 ( Clx3, CPA+E2 ) on mechanical paw withdrawal threshold in the rat.
    Figure Legend Snippet: A , Repeated administration of clonidine produces tolerance to antinociception. Effect of PGE 2 ( E2 ), clonidine plus PGE 2 ( Cl+E2 ), clonidine once hourly for 3 hr ( Clx3 ), clonidine once hourly for 3 hr, and at the fourth hour clonidine plus PGE 2 ( Clx3, Cl+E2 ) on mechanical paw withdrawal threshold in the rat. B , Bidirectional cross-tolerance develops among A 1 , α 2 , and μ antinociception. Effect of clonidine plus PGE 2 ( Cl+E2 ), DAMGO once hourly for 3 hr ( Dx3 ), DAMGO once hourly for 3 hr and at the fourth hour clonidine plus PGE 2 ( Dx3,Cl+E2 ), CPA once hourly for 3 hr and at the fourth hour clonidine plus PGE 2 ( CPAx3, Cl+E2 ), CPA once hourly for 3 hr ( CPAx3 ), clonidine once hourly for 3 hr and at the fourth hour DAMGO plus PGE 2 ( Clx3, D+E2 ), and clonidine once hourly for 3 hr and at the fourth hour CPA plus PGE 2 ( Clx3, CPA+E2 ) on mechanical paw withdrawal threshold in the rat.

    Techniques Used:

    μ, α 2 , and A 1 antagonists dose-dependently block μ, α 2 , and A 1 antinociception, respectively. A , Naloxone dose-dependently blocks DAMGO antinociception. Effect of PGE 2 ( E2 ), DAMGO plus PGE 2 ( D+E2 ) and various doses of naloxone (1 ng to 1 μg), and DAMGO plus PGE 2 ( N+D+E2 ), on mechanical paw withdrawal threshold in the rat. B , Yohimbine dose-dependently blocks clonidine antinociception. Effect of PGE 2 ( E2 ), clonidine plus PGE 2 ( Cl+E2 ), and various doses of yohimbine (1 ng to 1 μg) and clonidine plus PGE 2 ( Yo+Cl+E2 ) on mechanical paw withdrawal threshold in the rat. C , PACPX dose-dependently blocks CPA antinociception. Effect of PGE 2 ( E2 ), DAMGO plus PGE 2 ( CPA+E2 ), and various doses of PACPX (1 ng to 1 μg) and CPA plus PGE 2 ( PACPX+CPA+E2 ) on mechanical paw withdrawal threshold in the rat.
    Figure Legend Snippet: μ, α 2 , and A 1 antagonists dose-dependently block μ, α 2 , and A 1 antinociception, respectively. A , Naloxone dose-dependently blocks DAMGO antinociception. Effect of PGE 2 ( E2 ), DAMGO plus PGE 2 ( D+E2 ) and various doses of naloxone (1 ng to 1 μg), and DAMGO plus PGE 2 ( N+D+E2 ), on mechanical paw withdrawal threshold in the rat. B , Yohimbine dose-dependently blocks clonidine antinociception. Effect of PGE 2 ( E2 ), clonidine plus PGE 2 ( Cl+E2 ), and various doses of yohimbine (1 ng to 1 μg) and clonidine plus PGE 2 ( Yo+Cl+E2 ) on mechanical paw withdrawal threshold in the rat. C , PACPX dose-dependently blocks CPA antinociception. Effect of PGE 2 ( E2 ), DAMGO plus PGE 2 ( CPA+E2 ), and various doses of PACPX (1 ng to 1 μg) and CPA plus PGE 2 ( PACPX+CPA+E2 ) on mechanical paw withdrawal threshold in the rat.

    Techniques Used: Blocking Assay

    Multiple receptors are involved in μ, α 2 , and A 1 antinociception. A , Clonidine α 2 antinociception is blocked not only by yohimbine but also by naloxone and PACPX. Effect of PGE 2 ( E2 ), clonidine plus PGE 2 ( Cl+E2 ), yohimbine plus clonidine plus PGE 2 ( Yo+Cl+E2 ), naloxone plus clonidine plus PGE 2 ( N+Cl+E2 ), and PACPX plus clonidine plus PGE 2 ( PACPX+Cl+E2 ) on mechanical paw withdrawal threshold in the rat. B , DAMGO μ antinociception is blocked not only by naloxone but also by yohimbine. Effect of PGE 2 ( E2 ), DAMGO plus PGE 2 ( D+E2 ), naloxone plus DAMGO plus PGE 2 ( N+D+E2 ), yohimbine plus DAMGO plus PGE 2 ( Yo+D+E2 ), and PACPX plus DAMGO plus PGE 2 ( PACPX+D+E2 ) on mechanical paw withdrawal threshold in the rat. C , CPA A 1 antinociception is blocked not only by PACPX but also by yohimbine. Effect of PGE 2 ( E2 ), CPA plus PGE 2 ( CPA+E2 ), PACPX plus CPA plus PGE 2 ( PACPX+CPA+E2 ), naloxone plus CPA plus PGE 2 ( N+CPA+E2 ), and yohimbine plus CPA plus PGE 2 ( Yo+CPA+E2 ) on mechanical paw withdrawal threshold in the rat.
    Figure Legend Snippet: Multiple receptors are involved in μ, α 2 , and A 1 antinociception. A , Clonidine α 2 antinociception is blocked not only by yohimbine but also by naloxone and PACPX. Effect of PGE 2 ( E2 ), clonidine plus PGE 2 ( Cl+E2 ), yohimbine plus clonidine plus PGE 2 ( Yo+Cl+E2 ), naloxone plus clonidine plus PGE 2 ( N+Cl+E2 ), and PACPX plus clonidine plus PGE 2 ( PACPX+Cl+E2 ) on mechanical paw withdrawal threshold in the rat. B , DAMGO μ antinociception is blocked not only by naloxone but also by yohimbine. Effect of PGE 2 ( E2 ), DAMGO plus PGE 2 ( D+E2 ), naloxone plus DAMGO plus PGE 2 ( N+D+E2 ), yohimbine plus DAMGO plus PGE 2 ( Yo+D+E2 ), and PACPX plus DAMGO plus PGE 2 ( PACPX+D+E2 ) on mechanical paw withdrawal threshold in the rat. C , CPA A 1 antinociception is blocked not only by PACPX but also by yohimbine. Effect of PGE 2 ( E2 ), CPA plus PGE 2 ( CPA+E2 ), PACPX plus CPA plus PGE 2 ( PACPX+CPA+E2 ), naloxone plus CPA plus PGE 2 ( N+CPA+E2 ), and yohimbine plus CPA plus PGE 2 ( Yo+CPA+E2 ) on mechanical paw withdrawal threshold in the rat.

    Techniques Used:

    Antisense α 2C ODN treatment blocks not only α 2 antinociception but also μ and A 1 antinociception. A , Effect of PGE 2 ( E2 ), clonidine plus PGE 2 ( Cl+E2 ), α 2 -(AS) ODN 1 μg intrathecally on alternate days × 3, and clonidine plus PGE 2 [ α 2 -(AS)x3,CCl+E2 ], α 2 -(S) ODN 1 μg intrathecally on alternate days × 3, and clonidine plus PGE 2 [ α 2 -(S)x3,Cl+E2 ] on mechanical paw withdrawal threshold in the rat. B , Effect of PGE 2 ( E2 ), DAMGO plus PGE 2 ( D+E2 ), α 2 -(AS) ODN 1 μg intrathecally on alternate days × 3, and DAMGO plus PGE 2 [ α 2 -(AS)x3,D+E2 ], α 2 -(S) ODN 1 μg intrathecally on alternate days × 3, and DAMGO plus PGE 2 [ α 2 -(S)x3,D+E2 ] on mechanical paw withdrawal threshold in the rat. C , Effect of PGE 2 (E2), CPA plus PGE 2 (CPA + E2), α 2 -(AS) ODN 1 μg intrathecally on alternate days × 3, and CPA plus PGE 2 [ α 2 -(AS)x3,CPA+E2 ], α 2 -(S) ODN 1 μg intrathecally on alternate days × 3, and CPA plus PGE 2 [ α 2 -(S)x3,CPA+E2 ] on mechanical paw withdrawal threshold in the rat.
    Figure Legend Snippet: Antisense α 2C ODN treatment blocks not only α 2 antinociception but also μ and A 1 antinociception. A , Effect of PGE 2 ( E2 ), clonidine plus PGE 2 ( Cl+E2 ), α 2 -(AS) ODN 1 μg intrathecally on alternate days × 3, and clonidine plus PGE 2 [ α 2 -(AS)x3,CCl+E2 ], α 2 -(S) ODN 1 μg intrathecally on alternate days × 3, and clonidine plus PGE 2 [ α 2 -(S)x3,Cl+E2 ] on mechanical paw withdrawal threshold in the rat. B , Effect of PGE 2 ( E2 ), DAMGO plus PGE 2 ( D+E2 ), α 2 -(AS) ODN 1 μg intrathecally on alternate days × 3, and DAMGO plus PGE 2 [ α 2 -(AS)x3,D+E2 ], α 2 -(S) ODN 1 μg intrathecally on alternate days × 3, and DAMGO plus PGE 2 [ α 2 -(S)x3,D+E2 ] on mechanical paw withdrawal threshold in the rat. C , Effect of PGE 2 (E2), CPA plus PGE 2 (CPA + E2), α 2 -(AS) ODN 1 μg intrathecally on alternate days × 3, and CPA plus PGE 2 [ α 2 -(AS)x3,CPA+E2 ], α 2 -(S) ODN 1 μg intrathecally on alternate days × 3, and CPA plus PGE 2 [ α 2 -(S)x3,CPA+E2 ] on mechanical paw withdrawal threshold in the rat.

    Techniques Used:

    25) Product Images from "Drug Dependent Sex Differences in Periaqueducatal Gray Mediated Antinociception in the Rat"

    Article Title: Drug Dependent Sex Differences in Periaqueducatal Gray Mediated Antinociception in the Rat

    Journal: Pain

    doi: 10.1016/j.pain.2009.09.008

    Model of PAG circuitry underlying antinociception. Administration of morphine, DAMGO, fentanyl, bicuculline, or kainic acid causes excitation of vlPAG output neurons. MOPr agonists inhibit GABAergic neurons, which disinhibits vlPAG output neurons projecting
    Figure Legend Snippet: Model of PAG circuitry underlying antinociception. Administration of morphine, DAMGO, fentanyl, bicuculline, or kainic acid causes excitation of vlPAG output neurons. MOPr agonists inhibit GABAergic neurons, which disinhibits vlPAG output neurons projecting

    Techniques Used:

    26) Product Images from "Mu-opioid receptor and delta-opioid receptor differentially regulate microglial inflammatory response to control proopiomelanocortin neuronal apoptosis in the hypothalamus: effects of neonatal alcohol"

    Article Title: Mu-opioid receptor and delta-opioid receptor differentially regulate microglial inflammatory response to control proopiomelanocortin neuronal apoptosis in the hypothalamus: effects of neonatal alcohol

    Journal: Journal of Neuroinflammation

    doi: 10.1186/s12974-017-0844-3

    The effect of opioid agonists and antagonists and immunoneutralization of inflammatory cytokines TNF-α and IL-6 on the ability of ethanol-or opioid-activated microglial conditioned media to induce apoptosis of POMC neurons. POMC neurons were differentiated from neural stem cells in culture and maintained in T25 flasks (1 × 10 6 /well) for 2 days and then treated for 24 h with microglial conditioned media exposed to opioidergic agents for 24 h before determining neuronal apoptosis using a nucleosome assay. Bar graphs are showing the apoptotic effects of ethanol (50 mM) with or without DAMGO (50 μM), naltrexone (10 ng/ml), or DAMGO and naltrexone ( a ); ethanol (50 mM) with or without DPDPE (10 nM), naltrindole (50 μM), or DPDPE and naltrindole ( b ). Microglial conditioned media from ethanol with or without opioidergic agonist-treated cultures were mixed with antibody to TNF-α (1 ng/ml ( c )), antibody to IL-6 (0.5 ng/ml ( d )), antibody to IL-4 (1 ng/ml ( e )), or antibody to IL-13 (5 ng/ml ( f )) and added to POMC neuron cultures for 24 h to determine neuronal apoptosis. The effects of immunoneutralization of inflammatory and anti-inflammatory cytokines on ethanol with or without opioid-activated POMC neuronal apoptosis are shown in bar graphs. Each bar represents mean ± SEM of 5–8 samples. Data were compared by one-way analysis of variance (ANOVA) and the Newman-Keuls posttest. Differences between groups are shown by lines with p values on the top of bar graphs
    Figure Legend Snippet: The effect of opioid agonists and antagonists and immunoneutralization of inflammatory cytokines TNF-α and IL-6 on the ability of ethanol-or opioid-activated microglial conditioned media to induce apoptosis of POMC neurons. POMC neurons were differentiated from neural stem cells in culture and maintained in T25 flasks (1 × 10 6 /well) for 2 days and then treated for 24 h with microglial conditioned media exposed to opioidergic agents for 24 h before determining neuronal apoptosis using a nucleosome assay. Bar graphs are showing the apoptotic effects of ethanol (50 mM) with or without DAMGO (50 μM), naltrexone (10 ng/ml), or DAMGO and naltrexone ( a ); ethanol (50 mM) with or without DPDPE (10 nM), naltrindole (50 μM), or DPDPE and naltrindole ( b ). Microglial conditioned media from ethanol with or without opioidergic agonist-treated cultures were mixed with antibody to TNF-α (1 ng/ml ( c )), antibody to IL-6 (0.5 ng/ml ( d )), antibody to IL-4 (1 ng/ml ( e )), or antibody to IL-13 (5 ng/ml ( f )) and added to POMC neuron cultures for 24 h to determine neuronal apoptosis. The effects of immunoneutralization of inflammatory and anti-inflammatory cytokines on ethanol with or without opioid-activated POMC neuronal apoptosis are shown in bar graphs. Each bar represents mean ± SEM of 5–8 samples. Data were compared by one-way analysis of variance (ANOVA) and the Newman-Keuls posttest. Differences between groups are shown by lines with p values on the top of bar graphs

    Techniques Used:

    27) Product Images from "Gi/o-Coupled Receptors Compete for Signaling to Adenylyl Cyclase in SH-SY5Y Cells and Reduce Opioid-Mediated cAMP Overshoot"

    Article Title: Gi/o-Coupled Receptors Compete for Signaling to Adenylyl Cyclase in SH-SY5Y Cells and Reduce Opioid-Mediated cAMP Overshoot

    Journal: Molecular Pharmacology

    doi: 10.1124/mol.110.064816

    Inhibition of cAMP by MOR or DOR agonists is similar for sensitized or nonsensitized AC. A, SH-SY5Y cells were incubated with vehicle (■) or the MOR agonist DAMGO (100 nM, □) overnight to induce dependence. Withdrawal was precipitated with the MOR antagonist CTAP (1 μM) in the presence of 5 μM forskolin. Short-term cAMP production was inhibited by including various concentrations of the DOR agonist SNC80 in the precipitating media. The concentration-response of SNC80 to inhibit cAMP was similar in control and DAMGO-dependent cells (EC 50 : vehicle-treated = 14.6 ± 7.8 nM, DAMGO-treated, 13.8 ± 7.2 nM; p > 0.05 by two-tailed student's t test). B, cells were incubated with vehicle (■) or the DOR agonist DPDPE (10 μM, □) overnight to induce dependence. Receptor-specific withdrawal was precipitated with the DOR antagonist ICI 174,864 (1 μM) in the presence of 30 μM forskolin. Various concentrations of the MOR agonist DAMGO were included in the precipitating media. The concentration-response of DAMGO to inhibit cAMP production was similar in control and DPDPE-withdrawn cells (EC 50 : vehicle-treated, 32.2 ± 12.5 nM, DPDPE-treated, 29.0 ± 7.1 nM; p > 0.05 by two-tailed Student's t test). Data are presented as mean picomoles of cAMP per milligram of protein ± S.E.M. ( n = 3 or 4, in duplicate). cAMP produced by forskolin alone is indicated by the dashed line.
    Figure Legend Snippet: Inhibition of cAMP by MOR or DOR agonists is similar for sensitized or nonsensitized AC. A, SH-SY5Y cells were incubated with vehicle (■) or the MOR agonist DAMGO (100 nM, □) overnight to induce dependence. Withdrawal was precipitated with the MOR antagonist CTAP (1 μM) in the presence of 5 μM forskolin. Short-term cAMP production was inhibited by including various concentrations of the DOR agonist SNC80 in the precipitating media. The concentration-response of SNC80 to inhibit cAMP was similar in control and DAMGO-dependent cells (EC 50 : vehicle-treated = 14.6 ± 7.8 nM, DAMGO-treated, 13.8 ± 7.2 nM; p > 0.05 by two-tailed student's t test). B, cells were incubated with vehicle (■) or the DOR agonist DPDPE (10 μM, □) overnight to induce dependence. Receptor-specific withdrawal was precipitated with the DOR antagonist ICI 174,864 (1 μM) in the presence of 30 μM forskolin. Various concentrations of the MOR agonist DAMGO were included in the precipitating media. The concentration-response of DAMGO to inhibit cAMP production was similar in control and DPDPE-withdrawn cells (EC 50 : vehicle-treated, 32.2 ± 12.5 nM, DPDPE-treated, 29.0 ± 7.1 nM; p > 0.05 by two-tailed Student's t test). Data are presented as mean picomoles of cAMP per milligram of protein ± S.E.M. ( n = 3 or 4, in duplicate). cAMP produced by forskolin alone is indicated by the dashed line.

    Techniques Used: Inhibition, Incubation, Concentration Assay, Two Tailed Test, Produced

    DAMGO-mediated cAMP overshoot is reduced by heterologous inhibition of shared AC by agonist to DOR, NOPr, or α 2 AR. AC sensitization was developed by incubating SH-SY5Y cells overnight with 100 nM DAMGO. To precipitate withdrawal, DAMGO-containing media were replaced with media containing 5 μM forskolin, 1 mM IBMX, and 1 μM CTAP in the presence or absence of 1 μM concentration of a non-MOR agonist, as indicated, for 10 min. Data are presented as mean ± S.E.M. of the percentage of cAMP overshoot, where stimulation by forskolin alone is represented as 0%. Three of six experiments, in duplicate, were compiled that produced > 100% DAMGO overshoot in the absence of a non-MOR agonist. **, p
    Figure Legend Snippet: DAMGO-mediated cAMP overshoot is reduced by heterologous inhibition of shared AC by agonist to DOR, NOPr, or α 2 AR. AC sensitization was developed by incubating SH-SY5Y cells overnight with 100 nM DAMGO. To precipitate withdrawal, DAMGO-containing media were replaced with media containing 5 μM forskolin, 1 mM IBMX, and 1 μM CTAP in the presence or absence of 1 μM concentration of a non-MOR agonist, as indicated, for 10 min. Data are presented as mean ± S.E.M. of the percentage of cAMP overshoot, where stimulation by forskolin alone is represented as 0%. Three of six experiments, in duplicate, were compiled that produced > 100% DAMGO overshoot in the absence of a non-MOR agonist. **, p

    Techniques Used: Inhibition, Concentration Assay, Produced

    MOR and DOR share AC during long-term agonist administration. SH-SY5Y cells were treated overnight with vehicle (□) or 10 μM DPDPE ( ) in the presence or absence of the MOR agonist DAMGO (10 or 100 nM) to induce dependence. Withdrawal was precipitated with the opioid antagonist naloxone (100 μM) in the presence of 5 μM forskolin. Data are presented as mean ± S.E.M. ( n = 4, in duplicate) of the percentage of forskolin-stimulated cAMP, where forskolin alone is 100% and is indicated by the dashed line. Overnight incubation with DPDPE produced overshoot on its own and enhanced the overshoot produced by 10 nM but not 100 nM DAMGO. ***, p
    Figure Legend Snippet: MOR and DOR share AC during long-term agonist administration. SH-SY5Y cells were treated overnight with vehicle (□) or 10 μM DPDPE ( ) in the presence or absence of the MOR agonist DAMGO (10 or 100 nM) to induce dependence. Withdrawal was precipitated with the opioid antagonist naloxone (100 μM) in the presence of 5 μM forskolin. Data are presented as mean ± S.E.M. ( n = 4, in duplicate) of the percentage of forskolin-stimulated cAMP, where forskolin alone is 100% and is indicated by the dashed line. Overnight incubation with DPDPE produced overshoot on its own and enhanced the overshoot produced by 10 nM but not 100 nM DAMGO. ***, p

    Techniques Used: Incubation, Produced

    28) Product Images from "Effect of μ-opioid agonist DAMGO on surface CXCR4 and HIV-1 replication in TF-1 human bone marrow progenitor cells"

    Article Title: Effect of μ-opioid agonist DAMGO on surface CXCR4 and HIV-1 replication in TF-1 human bone marrow progenitor cells

    Journal: BMC Research Notes

    doi: 10.1186/1756-0500-7-752

    DAMGO treatment inhibits the replication of HIV-1 X4-utilizing strain IIIB in TF-1 cells. Analyses involving the detection of HIV-1 p24 capsid protein were performed on cellular supernatants 24 hours after infection to assess HIV-1 IIIB replication in response to DAMGO treatment of TF-1 cells. Absolute values of p24 levels are denoted in ng/ml on the Y axis. A 2.3-fold decrease was observed with DAMGO treatment (1 μM), which further declined to 3.5-fold at 10 μM. CTAP pretreatment reversed DAMGO-mediated decline in p24 levels. Samples were assayed in triplicate and results shown are the average of two independent experiments. *p = 0.00012, **p = 0.00000076, and *** p = 0.000002.
    Figure Legend Snippet: DAMGO treatment inhibits the replication of HIV-1 X4-utilizing strain IIIB in TF-1 cells. Analyses involving the detection of HIV-1 p24 capsid protein were performed on cellular supernatants 24 hours after infection to assess HIV-1 IIIB replication in response to DAMGO treatment of TF-1 cells. Absolute values of p24 levels are denoted in ng/ml on the Y axis. A 2.3-fold decrease was observed with DAMGO treatment (1 μM), which further declined to 3.5-fold at 10 μM. CTAP pretreatment reversed DAMGO-mediated decline in p24 levels. Samples were assayed in triplicate and results shown are the average of two independent experiments. *p = 0.00012, **p = 0.00000076, and *** p = 0.000002.

    Techniques Used: Infection

    29) Product Images from "SEIZURE ACTIVITY INVOLVED IN THE UP-REGULATION OF BDNF mRNA EXPRESSION BY ACTIVATION OF CENTRAL MU OPIOID RECEPTORS"

    Article Title: SEIZURE ACTIVITY INVOLVED IN THE UP-REGULATION OF BDNF mRNA EXPRESSION BY ACTIVATION OF CENTRAL MU OPIOID RECEPTORS

    Journal: Neuroscience

    doi: 10.1016/j.neuroscience.2009.03.020

    Effects of systemic administration of diazepam (10 mg/kg, i.m.), phenobarbital (40 mg/kg, i.p.), and valproate (300 mg/kg, i.p.) on the behavioral seizure score increased by i.c.v. morphine 100 μg or i.c.v. DAMGO 1.5 μg. Diazepam was administered 15 min before administration of morphine or DAMGO. Phenobarbital or valproate was administered 1 h before administration of morphine or DAMGO. Each animal was scored according to the highest class of behavioral manifestation observed during the 1 h observation period. Each value represents mean ± S.E.M. (n=6).**, P
    Figure Legend Snippet: Effects of systemic administration of diazepam (10 mg/kg, i.m.), phenobarbital (40 mg/kg, i.p.), and valproate (300 mg/kg, i.p.) on the behavioral seizure score increased by i.c.v. morphine 100 μg or i.c.v. DAMGO 1.5 μg. Diazepam was administered 15 min before administration of morphine or DAMGO. Phenobarbital or valproate was administered 1 h before administration of morphine or DAMGO. Each animal was scored according to the highest class of behavioral manifestation observed during the 1 h observation period. Each value represents mean ± S.E.M. (n=6).**, P

    Techniques Used:

    30) Product Images from "Electroacupuncture Attenuates 5?-Guanidinonaltrindole-Evoked Scratching and Spinal c-Fos Expression in the Mouse"

    Article Title: Electroacupuncture Attenuates 5?-Guanidinonaltrindole-Evoked Scratching and Spinal c-Fos Expression in the Mouse

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    doi: 10.1155/2013/319124

    Effects of DAMGO by i.d. injection (a) and by s.c. injection (b) on GNTI- (0.3 mg/kg) induced scratching. (a) Mice were administered an i.d. injection of saline or DAMGO (10 nmol) at 15 min before GNTI (0.3 mg/kg). (b) Mice were administered a s.c. injection of saline or DAMGO (10 nmol) at 15 min before GNTI (0.3 mg/kg). The number of scratches was counted for 30 min after GNTI injection. DAMGO (10 nmol/site) attenuated GNTI-induced scratch behavior by i.d. administration but not s.c. administration. (* P
    Figure Legend Snippet: Effects of DAMGO by i.d. injection (a) and by s.c. injection (b) on GNTI- (0.3 mg/kg) induced scratching. (a) Mice were administered an i.d. injection of saline or DAMGO (10 nmol) at 15 min before GNTI (0.3 mg/kg). (b) Mice were administered a s.c. injection of saline or DAMGO (10 nmol) at 15 min before GNTI (0.3 mg/kg). The number of scratches was counted for 30 min after GNTI injection. DAMGO (10 nmol/site) attenuated GNTI-induced scratch behavior by i.d. administration but not s.c. administration. (* P

    Techniques Used: Injection, Mouse Assay

    31) Product Images from "Brain region- and sex-specific alterations in DAMGO-stimulated [35S]GTPγS binding in mice with Oprm1 A112G"

    Article Title: Brain region- and sex-specific alterations in DAMGO-stimulated [35S]GTPγS binding in mice with Oprm1 A112G

    Journal: Addiction biology

    doi: 10.1111/j.1369-1600.2012.00484.x

    Brain regions that show significant sex differences in DAMGO-stimulated [ 35 S]GTPγS binding in G/G mice
    Figure Legend Snippet: Brain regions that show significant sex differences in DAMGO-stimulated [ 35 S]GTPγS binding in G/G mice

    Techniques Used: Binding Assay, Mouse Assay

    DAMGO-stimulated [35 S]GTPγS binding in A/A and G/G mouse brains
    Figure Legend Snippet: DAMGO-stimulated [35 S]GTPγS binding in A/A and G/G mouse brains

    Techniques Used: Binding Assay

    Pseudo-color autoradiograms of DAMGO-stimulated [ 35 S]GTPγS binding to the MOPR in selected coronal sections of A/A and G/G male mouse brains at 3 anatomical levels (A–C)
    Figure Legend Snippet: Pseudo-color autoradiograms of DAMGO-stimulated [ 35 S]GTPγS binding to the MOPR in selected coronal sections of A/A and G/G male mouse brains at 3 anatomical levels (A–C)

    Techniques Used: Binding Assay

    In the nucleus accumbens core, there is significant genotype difference in DAMGO-stimulated [ 35 S]GTPγS binding in female mice
    Figure Legend Snippet: In the nucleus accumbens core, there is significant genotype difference in DAMGO-stimulated [ 35 S]GTPγS binding in female mice

    Techniques Used: Binding Assay, Mouse Assay

    Autoradiograms of DAMGO-stimulated [ 35 S]GTPγS binding in selected coronal sections of male A/A mouse brains at the telencephalon (A), diencephalon (B) and midbrain (C) levels
    Figure Legend Snippet: Autoradiograms of DAMGO-stimulated [ 35 S]GTPγS binding in selected coronal sections of male A/A mouse brains at the telencephalon (A), diencephalon (B) and midbrain (C) levels

    Techniques Used: Binding Assay

    32) Product Images from "Dopamine Depletion Reorganizes Projections from the Nucleus Accumbens and Ventral Pallidum That Mediate Opioid-Induced Motor Activity"

    Article Title: Dopamine Depletion Reorganizes Projections from the Nucleus Accumbens and Ventral Pallidum That Mediate Opioid-Induced Motor Activity

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.18-19-08074.1998

    Effect of fluphenazine (9.5 nmol/0.5 μl) microinjections in the nucleus accumbens 5 min before DAMGO microinjections in the nucleus accumbens. The data are shown as mean ± SEM photocell counts (horizontal or rearing activity) or centimeters
    Figure Legend Snippet: Effect of fluphenazine (9.5 nmol/0.5 μl) microinjections in the nucleus accumbens 5 min before DAMGO microinjections in the nucleus accumbens. The data are shown as mean ± SEM photocell counts (horizontal or rearing activity) or centimeters

    Techniques Used: Activity Assay

    Effect of muscimol (0.02 nmol/0.5 μl) in the VP on the motor responses elicited by DAMGO (0.3 nmol/0.5 μl) microinjection into the nucleus accumbens of 6-OHDA-lesioned ( n = 8) or sham-lesioned ( n = 6) rats. Behavior , Left , The data are
    Figure Legend Snippet: Effect of muscimol (0.02 nmol/0.5 μl) in the VP on the motor responses elicited by DAMGO (0.3 nmol/0.5 μl) microinjection into the nucleus accumbens of 6-OHDA-lesioned ( n = 8) or sham-lesioned ( n = 6) rats. Behavior , Left , The data are

    Techniques Used:

    Effect of baclofen (0.1 nmol/0.5 μl) in the VTA on the motor responses elicited by DAMGO (0.3 nmol/0.5 μl) microinjection into the nucleus accumbens. Behavior , Left , The data are shown as mean ± SEM photocell counts (horizontal
    Figure Legend Snippet: Effect of baclofen (0.1 nmol/0.5 μl) in the VTA on the motor responses elicited by DAMGO (0.3 nmol/0.5 μl) microinjection into the nucleus accumbens. Behavior , Left , The data are shown as mean ± SEM photocell counts (horizontal

    Techniques Used:

    Effect of baclofen (0.1 nmol/0.5 μl) in the VTA on the motor responses elicited by DAMGO (0.3 nmol/0.5 μl) microinjection into the VP. Behavior , Left , The data are shown as mean ± SEM photocell counts (horizontal or rearing activity)
    Figure Legend Snippet: Effect of baclofen (0.1 nmol/0.5 μl) in the VTA on the motor responses elicited by DAMGO (0.3 nmol/0.5 μl) microinjection into the VP. Behavior , Left , The data are shown as mean ± SEM photocell counts (horizontal or rearing activity)

    Techniques Used: Activity Assay

    33) Product Images from "Desensitization of ?-opioid receptor-evoked potassium currents: Initiation at the receptor, expression at the effector"

    Article Title: Desensitization of ?-opioid receptor-evoked potassium currents: Initiation at the receptor, expression at the effector

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi: 10.1073/pnas.072075399

    Desensitized DAMGO-elicited GIRK currents may be reactivated. ( A ) Stimulating α 2 ARs at the peak amplitude of the DAMGO-mediated response did not activate any additional GIRK current, indicating that both receptor systems share the same population of GIRK channels. ( B ) Stimulating α 2 ARs after desensitization of the DAMGO-mediated response partially restored the initial GIRK current amplitude. (Scale bars in A and B , 100 pA and 5 min; V h = −73 mV.) ( C ) Bar graph representation of mean (±SE) normalized current amplitudes evoked by α 2 ARs stimulation (light gray bars) or SST (dark gray bar) stacked on bars representing MOR-elicited responses (empty bars) at the peak or in the desensitized state. Note that addition of SST onto DAMGO plus UK further increases the reactivation of the desensitized GIRK currents. Responses are expressed as percent of the peak DAMGO-elicited response in the same cell ( n = 4 for each condition). UK, 3 μM; Yoh, 10 μM; DAMGO, 1 μM; Nal, 1 μM.
    Figure Legend Snippet: Desensitized DAMGO-elicited GIRK currents may be reactivated. ( A ) Stimulating α 2 ARs at the peak amplitude of the DAMGO-mediated response did not activate any additional GIRK current, indicating that both receptor systems share the same population of GIRK channels. ( B ) Stimulating α 2 ARs after desensitization of the DAMGO-mediated response partially restored the initial GIRK current amplitude. (Scale bars in A and B , 100 pA and 5 min; V h = −73 mV.) ( C ) Bar graph representation of mean (±SE) normalized current amplitudes evoked by α 2 ARs stimulation (light gray bars) or SST (dark gray bar) stacked on bars representing MOR-elicited responses (empty bars) at the peak or in the desensitized state. Note that addition of SST onto DAMGO plus UK further increases the reactivation of the desensitized GIRK currents. Responses are expressed as percent of the peak DAMGO-elicited response in the same cell ( n = 4 for each condition). UK, 3 μM; Yoh, 10 μM; DAMGO, 1 μM; Nal, 1 μM.

    Techniques Used:

    Heterologous effects of MOR-induced desensitization. ( A ) Example of responses elicited successively by UK and DAMGO. Stimulation of α2ARs triggers a large outward current reversed by Yoh. Subsequent stimulation of MORs triggers an outward current of similar amplitude that desensitizes substantially within 15 min. ( B ) The application of the above agonists in the reversed order also elicited outward currents of similar amplitude, provided that DAMGO was applied only briefly. ( C ) However, if DAMGO is applied for 15 min and Nal used to reverse the response after desensitization occurred, subsequent application of UK elicited an outward current with a maximal amplitude similar to the preceding MOR-induced desensitized response. ( D ) Same as in C , except that addition of 3 μM SST onto the UK-induced response partially reactivated this heterologously desensitized response. ( A – D ) (Concentrations: DAMGO 1 μM, UK 3 μM, Nal 1 μM, Yoh 10 μM; scale bars, 100 pA and 10 min; V h = −73 mV.) ( E ) Bar graph representation of mean (±SE) normalized responses evoked by 3 μM UK ( n = 13), 3 μM somatostatin (SST; n = 3), or 100 μM Baclofen (GABA B agonist; n = 5) before 1 μM DAMGO for 15 min or conversely. The fifth bar from the left represents the responses to UK plus SST ( n = 4), reflecting partial reactivation of heterologous desensitization. Responses are expressed as percent of the peak DAMGO-elicited response in the same cell. *, P
    Figure Legend Snippet: Heterologous effects of MOR-induced desensitization. ( A ) Example of responses elicited successively by UK and DAMGO. Stimulation of α2ARs triggers a large outward current reversed by Yoh. Subsequent stimulation of MORs triggers an outward current of similar amplitude that desensitizes substantially within 15 min. ( B ) The application of the above agonists in the reversed order also elicited outward currents of similar amplitude, provided that DAMGO was applied only briefly. ( C ) However, if DAMGO is applied for 15 min and Nal used to reverse the response after desensitization occurred, subsequent application of UK elicited an outward current with a maximal amplitude similar to the preceding MOR-induced desensitized response. ( D ) Same as in C , except that addition of 3 μM SST onto the UK-induced response partially reactivated this heterologously desensitized response. ( A – D ) (Concentrations: DAMGO 1 μM, UK 3 μM, Nal 1 μM, Yoh 10 μM; scale bars, 100 pA and 10 min; V h = −73 mV.) ( E ) Bar graph representation of mean (±SE) normalized responses evoked by 3 μM UK ( n = 13), 3 μM somatostatin (SST; n = 3), or 100 μM Baclofen (GABA B agonist; n = 5) before 1 μM DAMGO for 15 min or conversely. The fifth bar from the left represents the responses to UK plus SST ( n = 4), reflecting partial reactivation of heterologous desensitization. Responses are expressed as percent of the peak DAMGO-elicited response in the same cell. *, P

    Techniques Used:

    34) Product Images from "Opioid-Induced Long-Term Potentiation in the Spinal Cord Is a Presynaptic Event"

    Article Title: Opioid-Induced Long-Term Potentiation in the Spinal Cord Is a Presynaptic Event

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

    doi: 10.1523/JNEUROSCI.5857-09.2010

    DAMGO induces LTP from primary afferent terminals in the spinal cord. ( A ) Blocking the postsynaptic opioid action with GDP-β-S and Cs 2+ failed to block LTP induction by DAMGO in 9 lamina II neurons without affecting the initial depression (black
    Figure Legend Snippet: DAMGO induces LTP from primary afferent terminals in the spinal cord. ( A ) Blocking the postsynaptic opioid action with GDP-β-S and Cs 2+ failed to block LTP induction by DAMGO in 9 lamina II neurons without affecting the initial depression (black

    Techniques Used: Blocking Assay

    Increased intracellular calcium is involved in opioid-induced presynaptic LTP in the spinal cord. ( A ) Postsynaptic dialysis with 30 mM BAPTA failed to block DAMGO (1 μM)-induced LTP in 9 lamina II neurons (black solid circles). DAMGO produced
    Figure Legend Snippet: Increased intracellular calcium is involved in opioid-induced presynaptic LTP in the spinal cord. ( A ) Postsynaptic dialysis with 30 mM BAPTA failed to block DAMGO (1 μM)-induced LTP in 9 lamina II neurons (black solid circles). DAMGO produced

    Techniques Used: Blocking Assay, Produced

    35) Product Images from "Different Mechanisms Mediate Development and Expression of Tolerance and Dependence for Peripheral μ-Opioid Antinociception in Rat"

    Article Title: Different Mechanisms Mediate Development and Expression of Tolerance and Dependence for Peripheral μ-Opioid Antinociception in Rat

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.17-20-08018.1997

    Calcium but not PKC or NO plays a role in the expression of peripheral opioid tolerance. Effects of PGE 2 , ( PGE 2 ; n = 16), DAMGO plus PGE 2 ( DAMGO+PGE 2 ; n = 16), three hourly injections of DAMGO followed by DAMGO plus PGE 2 ( DAMGO×3, DAMGO+PGE 2 ; n =
    Figure Legend Snippet: Calcium but not PKC or NO plays a role in the expression of peripheral opioid tolerance. Effects of PGE 2 , ( PGE 2 ; n = 16), DAMGO plus PGE 2 ( DAMGO+PGE 2 ; n = 16), three hourly injections of DAMGO followed by DAMGO plus PGE 2 ( DAMGO×3, DAMGO+PGE 2 ; n =

    Techniques Used: Expressing

    A , Role of NO in development of peripheral opioid tolerance. Effects of PGE 2 (100 ng; PGE 2 ; n = 16), DAMGO (1 μg) plus PGE 2 ( DAMGO+PGE 2 ; n = 16), three hourly injections of DAMGO followed by DAMGO plus PGE 2 at the fourth hour ( DAMGO×3,DAMGO+PGE
    Figure Legend Snippet: A , Role of NO in development of peripheral opioid tolerance. Effects of PGE 2 (100 ng; PGE 2 ; n = 16), DAMGO (1 μg) plus PGE 2 ( DAMGO+PGE 2 ; n = 16), three hourly injections of DAMGO followed by DAMGO plus PGE 2 at the fourth hour ( DAMGO×3,DAMGO+PGE

    Techniques Used:

    PKC but not NO, cAMP, or calcium plays a role in the development of peripheral opioid dependence. Effects of PGE 2 , ( PGE 2 ; n = 16), DAMGO plus PGE 2 ( DAMGO+PGE 2 ; n = 16), three hourly injections of DAMGO followed by vehicle at the fourth hour ( DAMGO×3,V
    Figure Legend Snippet: PKC but not NO, cAMP, or calcium plays a role in the development of peripheral opioid dependence. Effects of PGE 2 , ( PGE 2 ; n = 16), DAMGO plus PGE 2 ( DAMGO+PGE 2 ; n = 16), three hourly injections of DAMGO followed by vehicle at the fourth hour ( DAMGO×3,V

    Techniques Used:

    PKC, calcium, and cAMP, but not NO, play a role in expression of peripheral opioid dependence. Effects of PGE 2 ( PGE 2 ; n = 16), DAMGO plus PGE 2 ( DAMGO+PGE 2 ; n = 16), three hourly injections of DAMGO followed by naloxone at the fourth hour ( DAMGO×3,NAL
    Figure Legend Snippet: PKC, calcium, and cAMP, but not NO, play a role in expression of peripheral opioid dependence. Effects of PGE 2 ( PGE 2 ; n = 16), DAMGO plus PGE 2 ( DAMGO+PGE 2 ; n = 16), three hourly injections of DAMGO followed by naloxone at the fourth hour ( DAMGO×3,NAL

    Techniques Used: Expressing

    36) Product Images from "Opioids depress breathing through two small brainstem sites"

    Article Title: Opioids depress breathing through two small brainstem sites

    Journal: bioRxiv

    doi: 10.1101/807297

    Deletion of μ-Opiate receptor from neural subtypes to define their contribution to opioid depression of preBötC burst rhythm and amplitude. A, Schematic of single-cell mRNA sequencing paradigm. Postnatal day 0 (P0) medullary brainstem slices containing the preBötC bilaterally (circled in red) were dissected and isolated for sequencing. B , Single cell transcriptome profiling of cells isolated from P0 preBötC. Of 1817 cells isolated, only 267 were presumed preBötC neurons of which only 21 expressed Oprm1 mRNA. C , Heatmap of scaled transcript abundance for Oprm1 and markers of glutamatergic, gabaergic, and glycinergic preBötC neurons. Oprm1 expressing cells are both excitatory and inhibitory. D , Schematic of extracellular recordings of the preBötC rhythm in P0-4 medullary brainstem slices. The preBötC has input to the hypoglossal motor neurons which form the CN12 rootlet, relaying an inspiratory motor command to the tongue in intact animals. Due to its input from the preBötC, extracellular recording from this rootlet display autonomous rhythmic activity corresponding to in vitro respiration ( 5 ). E , Representative recording of bursting activity after application of 50nM DAMGO and 100nM Naloxone. Top: in control ( Oprm1 (f/f)) slices bath application of 50nM DAMGO quickly slowed and decreased the amplitude from baseline bursting. After rhythm cessation, bath application of Naloxone restored the rhythm. Bottom: in Vglut2 -cre; Oprm1 (f/f) mice 50nM DAMGO did not stop, or even slow rhythmic activity. F , Percent of slices from each genotype with rhythm cessation after 50nM DAMGO application. Control (n=11), Gad2 -Cre; Oprm1 (f/f) (n=6), Vgat -Cre; Oprm1 (f/f) (n=2), Glty2 -Cre; Oprm1 (f/f) (n=4), and Vglut2 -Cre; Oprm1 (f/f) (n=3). G , Schematic showing three subpopulations of glutamatergic lineages delineated by transcription factors Dbx1 and Foxp2 . Foxp2 neurons represents a smaller, and overlapping, population of Dbx1 neurons. H, Identification of molecular subtypes of Oprm1 preBötC excitatory neurons. Sagittal section of the preBötC from a P0 OPRM1::mCherry; Dbx1 -Cre;Rosa-LSL-YFP mouse immunostained for mCherry (red), YFP (Green) and FOXP2 (Blue). About ~50% of Oprm1 preBötC neurons are glutamatergic/ Dbx1 derived (arrowhead) and of those, ~35% express FOXP2(asterisk). Scale bar, 50μM. I, Quantification of the number of preBötC for each molecular subtype identified in H . J , Dose response curve for the bursting rate and amplitude after bath application of 0, 50, 100, and 500nM DAMGO applied to Vglut2 -Cre; Oprm1 (f/f) (gray, n=3), Dbx1 -Cre; Oprm1 (f/f) (green, n=3), Foxp2 -Cre; Oprm1 (f/f) (blue, n=4) and control (black, n=11) P0-4 preBötC slices. Rate and amplitude for each slice are normalized to baseline. K , Schematic summary showing that the key node for opioids to suppress breathing is the preBötC and within this site, elimination of Oprm1 from just a small subset of those neurons, ~70-140 excitatory neurons, prevents opioid respiratory suppression.
    Figure Legend Snippet: Deletion of μ-Opiate receptor from neural subtypes to define their contribution to opioid depression of preBötC burst rhythm and amplitude. A, Schematic of single-cell mRNA sequencing paradigm. Postnatal day 0 (P0) medullary brainstem slices containing the preBötC bilaterally (circled in red) were dissected and isolated for sequencing. B , Single cell transcriptome profiling of cells isolated from P0 preBötC. Of 1817 cells isolated, only 267 were presumed preBötC neurons of which only 21 expressed Oprm1 mRNA. C , Heatmap of scaled transcript abundance for Oprm1 and markers of glutamatergic, gabaergic, and glycinergic preBötC neurons. Oprm1 expressing cells are both excitatory and inhibitory. D , Schematic of extracellular recordings of the preBötC rhythm in P0-4 medullary brainstem slices. The preBötC has input to the hypoglossal motor neurons which form the CN12 rootlet, relaying an inspiratory motor command to the tongue in intact animals. Due to its input from the preBötC, extracellular recording from this rootlet display autonomous rhythmic activity corresponding to in vitro respiration ( 5 ). E , Representative recording of bursting activity after application of 50nM DAMGO and 100nM Naloxone. Top: in control ( Oprm1 (f/f)) slices bath application of 50nM DAMGO quickly slowed and decreased the amplitude from baseline bursting. After rhythm cessation, bath application of Naloxone restored the rhythm. Bottom: in Vglut2 -cre; Oprm1 (f/f) mice 50nM DAMGO did not stop, or even slow rhythmic activity. F , Percent of slices from each genotype with rhythm cessation after 50nM DAMGO application. Control (n=11), Gad2 -Cre; Oprm1 (f/f) (n=6), Vgat -Cre; Oprm1 (f/f) (n=2), Glty2 -Cre; Oprm1 (f/f) (n=4), and Vglut2 -Cre; Oprm1 (f/f) (n=3). G , Schematic showing three subpopulations of glutamatergic lineages delineated by transcription factors Dbx1 and Foxp2 . Foxp2 neurons represents a smaller, and overlapping, population of Dbx1 neurons. H, Identification of molecular subtypes of Oprm1 preBötC excitatory neurons. Sagittal section of the preBötC from a P0 OPRM1::mCherry; Dbx1 -Cre;Rosa-LSL-YFP mouse immunostained for mCherry (red), YFP (Green) and FOXP2 (Blue). About ~50% of Oprm1 preBötC neurons are glutamatergic/ Dbx1 derived (arrowhead) and of those, ~35% express FOXP2(asterisk). Scale bar, 50μM. I, Quantification of the number of preBötC for each molecular subtype identified in H . J , Dose response curve for the bursting rate and amplitude after bath application of 0, 50, 100, and 500nM DAMGO applied to Vglut2 -Cre; Oprm1 (f/f) (gray, n=3), Dbx1 -Cre; Oprm1 (f/f) (green, n=3), Foxp2 -Cre; Oprm1 (f/f) (blue, n=4) and control (black, n=11) P0-4 preBötC slices. Rate and amplitude for each slice are normalized to baseline. K , Schematic summary showing that the key node for opioids to suppress breathing is the preBötC and within this site, elimination of Oprm1 from just a small subset of those neurons, ~70-140 excitatory neurons, prevents opioid respiratory suppression.

    Techniques Used: Sequencing, Isolation, Expressing, Activity Assay, In Vitro, Mouse Assay, Derivative Assay

    preBötC slice activity in 50nM DAMGO after deletion of Oprm1 from inhibitory neural types. Representative recording of bursting activity at baseline, after application of 50nM DAMGO (red line), and 100nM Naloxone (blue line). Top: Vgat -Cre; Oprm1 (f/f) targeting GABAergic neurons (n=2). Middle: Gad2 -Cre; Oprm1 (f/f) targeting GABAergic neurons (n=6). Bottom: Glyt2 -Cre; Oprm1 (f/f) targeting Glycinergic neurons (n=4).
    Figure Legend Snippet: preBötC slice activity in 50nM DAMGO after deletion of Oprm1 from inhibitory neural types. Representative recording of bursting activity at baseline, after application of 50nM DAMGO (red line), and 100nM Naloxone (blue line). Top: Vgat -Cre; Oprm1 (f/f) targeting GABAergic neurons (n=2). Middle: Gad2 -Cre; Oprm1 (f/f) targeting GABAergic neurons (n=6). Bottom: Glyt2 -Cre; Oprm1 (f/f) targeting Glycinergic neurons (n=4).

    Techniques Used: Activity Assay

    37) Product Images from "Role of GPCR (Mu-Opioid)-RTK (Epidermal Growth Factor) Crosstalk in Opioid-Induced Hyperalgesic Priming (Type II)"

    Article Title: Role of GPCR (Mu-Opioid)-RTK (Epidermal Growth Factor) Crosstalk in Opioid-Induced Hyperalgesic Priming (Type II)

    Journal: Pain

    doi: 10.1097/j.pain.0000000000001155

    Schematic of mechanisms involved in induction of Type II hyperalgesic priming (OIH and prolongation of PGE 2 -induced hyperalgesia) Chronic opioid use changes MOR signaling, that may reflect intracellular signal switching. Our findings show that repeated (hourly × 4) intradermal injections of a MOR selective agonist DAMGO induces Type II hyperalgesic priming (OIH [ A ] and prolongation of PGE 2 -induced hyperalgesia [ B ]), in peptidergic IB4-negative nociceptors. Induction of both components present in Type II priming are MOR dependent, which activates diverse downstream second messengers. A. Activation of MOR (a Gα i -protein-coupled receptor) by repeated (hourly × 4) injections of DAMGO, stimulates Src and FAK, leading to activation of MAPK, which ultimately produces sensitization in the peripheral terminal of IB4-negative nociceptor. B. After the peripheral terminal of IB4-negative nociceptors have received repeated injections of DAMGO, PGE 2 was injected, which activates a signaling cascade, involving EGFR, Src and FAK, leading to stimulation of MAPK, which ultimately prolongs PGE 2 -induced hyperalgesia. Thus, while the induction of OIH is partially attenuated by the inhibition of Src, FAK and MAPK signaling ( A ), the prolongation of PGE 2 hyperalgesia is completely dependent on Src, FAK, EGFR and MAPK signaling ( B ). Schematics summarize the signaling pathways involved in the induction of OIH ( A ) and prolongation of PGE 2 hyperalgesia ( B ) induced by repeated exposure to DAMGO. Abbreviations : βγ, G-protein βγ subunit; DAMGO, [D-Ala 2 , NMe-Phe 4 , Gly-ol 5 ]-enkephalin acetate salt (a mu-opioid receptor agonist); EGFR, epidermal growth factor receptor; EP, prostaglandin receptor; FAK, focal adhesion kinase; Gα i , G-protein α i subunit; IB4, isolectin B4; MAPK, mitogen-activated protein kinase; MOR, mu-opioid receptor; PGE 2 , prostaglandin-E 2 ; Src, proto-oncogene tyrosine-protein kinase.
    Figure Legend Snippet: Schematic of mechanisms involved in induction of Type II hyperalgesic priming (OIH and prolongation of PGE 2 -induced hyperalgesia) Chronic opioid use changes MOR signaling, that may reflect intracellular signal switching. Our findings show that repeated (hourly × 4) intradermal injections of a MOR selective agonist DAMGO induces Type II hyperalgesic priming (OIH [ A ] and prolongation of PGE 2 -induced hyperalgesia [ B ]), in peptidergic IB4-negative nociceptors. Induction of both components present in Type II priming are MOR dependent, which activates diverse downstream second messengers. A. Activation of MOR (a Gα i -protein-coupled receptor) by repeated (hourly × 4) injections of DAMGO, stimulates Src and FAK, leading to activation of MAPK, which ultimately produces sensitization in the peripheral terminal of IB4-negative nociceptor. B. After the peripheral terminal of IB4-negative nociceptors have received repeated injections of DAMGO, PGE 2 was injected, which activates a signaling cascade, involving EGFR, Src and FAK, leading to stimulation of MAPK, which ultimately prolongs PGE 2 -induced hyperalgesia. Thus, while the induction of OIH is partially attenuated by the inhibition of Src, FAK and MAPK signaling ( A ), the prolongation of PGE 2 hyperalgesia is completely dependent on Src, FAK, EGFR and MAPK signaling ( B ). Schematics summarize the signaling pathways involved in the induction of OIH ( A ) and prolongation of PGE 2 hyperalgesia ( B ) induced by repeated exposure to DAMGO. Abbreviations : βγ, G-protein βγ subunit; DAMGO, [D-Ala 2 , NMe-Phe 4 , Gly-ol 5 ]-enkephalin acetate salt (a mu-opioid receptor agonist); EGFR, epidermal growth factor receptor; EP, prostaglandin receptor; FAK, focal adhesion kinase; Gα i , G-protein α i subunit; IB4, isolectin B4; MAPK, mitogen-activated protein kinase; MOR, mu-opioid receptor; PGE 2 , prostaglandin-E 2 ; Src, proto-oncogene tyrosine-protein kinase.

    Techniques Used: Activation Assay, Injection, Inhibition

    38) Product Images from "Morphine Stimulates Cell Migration of Oral Epithelial Cells by Delta-Opioid Receptor Activation"

    Article Title: Morphine Stimulates Cell Migration of Oral Epithelial Cells by Delta-Opioid Receptor Activation

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0042616

    Effect of selective opioid receptor agonists on the ‘wound closure’ in TR146 cells. Closed wound area % after 14 hours of exposure with selective opioid receptor agonist for MOR (DAMGO), KOR (U-69593) and DOR (DPDPE) (100 nM) in comparison to morphine (100 nM). TGF-ß (1 ng/ml) served as a positive control, (n = 3).***p≤0.001.
    Figure Legend Snippet: Effect of selective opioid receptor agonists on the ‘wound closure’ in TR146 cells. Closed wound area % after 14 hours of exposure with selective opioid receptor agonist for MOR (DAMGO), KOR (U-69593) and DOR (DPDPE) (100 nM) in comparison to morphine (100 nM). TGF-ß (1 ng/ml) served as a positive control, (n = 3).***p≤0.001.

    Techniques Used: Positive Control

    39) Product Images from "Pharmacological mechanisms underlying the antinociceptive and tolerance effects of the 6,14-bridged oripavine compound 030418"

    Article Title: Pharmacological mechanisms underlying the antinociceptive and tolerance effects of the 6,14-bridged oripavine compound 030418

    Journal: Acta Pharmacologica Sinica

    doi: 10.1038/aps.2011.83

    Effects of the nonselective opioid receptor antagonist naloxone (A), selective μ-opioid receptor antagonist β-FNA (B), selective κ-opioid receptor antagonist nor-BNI (C), and selective ORL1 receptor antagonist J-113397 (D) on the antinociception activity of 030418 in the mouse hot-plate tests. 030418 (4.0 μg/kg ABC; 2.0 μg/kg D, sc), morphine (6.0 mg/kg, sc), DAMGO (50 ng/mouse, icv), (±)U50488 (50 μg/mouse, icv), buprenorphine (1.7 mg/kg, sc), or vehicle control were administered before testing. The mice were pretreated with naloxone (1 mg/kg, sc), β-FNA (10 μg/mouse, icv), nor-BNI (3.7 μg/mouse, icv), or J-113397 (4.0 μg/mouse, icv) at 15 min, 24 h, 24 h, or 15 min, respectively, before administration of the corresponding agonists. The data are expressed as %MPE, and each point represents the mean±SEM ( n =8–10 mice). b P
    Figure Legend Snippet: Effects of the nonselective opioid receptor antagonist naloxone (A), selective μ-opioid receptor antagonist β-FNA (B), selective κ-opioid receptor antagonist nor-BNI (C), and selective ORL1 receptor antagonist J-113397 (D) on the antinociception activity of 030418 in the mouse hot-plate tests. 030418 (4.0 μg/kg ABC; 2.0 μg/kg D, sc), morphine (6.0 mg/kg, sc), DAMGO (50 ng/mouse, icv), (±)U50488 (50 μg/mouse, icv), buprenorphine (1.7 mg/kg, sc), or vehicle control were administered before testing. The mice were pretreated with naloxone (1 mg/kg, sc), β-FNA (10 μg/mouse, icv), nor-BNI (3.7 μg/mouse, icv), or J-113397 (4.0 μg/mouse, icv) at 15 min, 24 h, 24 h, or 15 min, respectively, before administration of the corresponding agonists. The data are expressed as %MPE, and each point represents the mean±SEM ( n =8–10 mice). b P

    Techniques Used: Activity Assay, Mouse Assay

    40) Product Images from "Desensitization-resistant and -sensitive GPCR-mediated inhibition of GABA release occurs by Ca2+-dependent and -independent mechanisms at a hypothalamic synapse"

    Article Title: Desensitization-resistant and -sensitive GPCR-mediated inhibition of GABA release occurs by Ca2+-dependent and -independent mechanisms at a hypothalamic synapse

    Journal: Journal of Neurophysiology

    doi: 10.1152/jn.00535.2015

    MOR- and GABA B R-mediated inhibition of release does not require the activation of K + channels. A : sample traces demonstrating either DAMGO- or baclofen-induced inhibition of eIPSC amplitude or spontaneous IPSC (sIPSC) frequency in the presence of the
    Figure Legend Snippet: MOR- and GABA B R-mediated inhibition of release does not require the activation of K + channels. A : sample traces demonstrating either DAMGO- or baclofen-induced inhibition of eIPSC amplitude or spontaneous IPSC (sIPSC) frequency in the presence of the

    Techniques Used: Inhibition, Activation Assay

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