dopamine d2r for co ip  (Alomone Labs)


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    Alomone Labs dopamine d2r for co ip
    The effects of D1-D2 heteromer stimulation and inactivation on basal conditioned place preference. (A) Vehicle-conditioned rats did not exhibit a preference toward a particular chamber. D1-D2 heteromer stimulation by SKF 83959 (1.5 mg/kg, s.c.) induced conditioned place aversion (CPA) as the animals spent significantly less time in the drug paired chamber. (B) SKF 83959-induced CPA was abolished by pre-treatment by the D1-D2 heteromer selective disrupting peptide, TAT-D1, but not the control TAT-Sc peptide. (C) Inactivation of D1-D2 heteromer by TAT-D1 resulted in conditioned place preference (CPP) as the rats spent significantly more time in the drug paired chamber, not observed with the control TAT-Sc. (D) Representative western blots (inset) and histogram showing the amount of D1R co-immunoprecipitated with <t>D2R</t> from the NAc of rats treated with saline or SKF 83959. Pretreatment with TAT-D1 led to decreased co-immunoprecipitated receptors. An aliquot of each sample was used as a control for WB (input control). (E,F) The CPA induced by D1-D2 heteromer stimulation was abolished by Cdk5 inhibitor roscovitine pre-treatment (200 nmol, i.c.v, E ) or intra-accumbal injection (30 nmol, F ). (G) Representative western blot and histogram showing the density of Thr75-DARPP-32 phosphorylation (pT75) relative to GAPDH (as loading control). Data represent means ± SEM of n = 8–10 rats/group. ( * p
    Dopamine D2r For Co Ip, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dopamine d2r for co ip/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    dopamine d2r for co ip - by Bioz Stars, 2022-07
    93/100 stars

    Images

    1) Product Images from "Activation of Dopamine D1-D2 Receptor Complex Attenuates Cocaine Reward and Reinstatement of Cocaine-Seeking through Inhibition of DARPP-32, ERK, and ΔFosB"

    Article Title: Activation of Dopamine D1-D2 Receptor Complex Attenuates Cocaine Reward and Reinstatement of Cocaine-Seeking through Inhibition of DARPP-32, ERK, and ΔFosB

    Journal: Frontiers in Pharmacology

    doi: 10.3389/fphar.2017.00924

    The effects of D1-D2 heteromer stimulation and inactivation on basal conditioned place preference. (A) Vehicle-conditioned rats did not exhibit a preference toward a particular chamber. D1-D2 heteromer stimulation by SKF 83959 (1.5 mg/kg, s.c.) induced conditioned place aversion (CPA) as the animals spent significantly less time in the drug paired chamber. (B) SKF 83959-induced CPA was abolished by pre-treatment by the D1-D2 heteromer selective disrupting peptide, TAT-D1, but not the control TAT-Sc peptide. (C) Inactivation of D1-D2 heteromer by TAT-D1 resulted in conditioned place preference (CPP) as the rats spent significantly more time in the drug paired chamber, not observed with the control TAT-Sc. (D) Representative western blots (inset) and histogram showing the amount of D1R co-immunoprecipitated with D2R from the NAc of rats treated with saline or SKF 83959. Pretreatment with TAT-D1 led to decreased co-immunoprecipitated receptors. An aliquot of each sample was used as a control for WB (input control). (E,F) The CPA induced by D1-D2 heteromer stimulation was abolished by Cdk5 inhibitor roscovitine pre-treatment (200 nmol, i.c.v, E ) or intra-accumbal injection (30 nmol, F ). (G) Representative western blot and histogram showing the density of Thr75-DARPP-32 phosphorylation (pT75) relative to GAPDH (as loading control). Data represent means ± SEM of n = 8–10 rats/group. ( * p
    Figure Legend Snippet: The effects of D1-D2 heteromer stimulation and inactivation on basal conditioned place preference. (A) Vehicle-conditioned rats did not exhibit a preference toward a particular chamber. D1-D2 heteromer stimulation by SKF 83959 (1.5 mg/kg, s.c.) induced conditioned place aversion (CPA) as the animals spent significantly less time in the drug paired chamber. (B) SKF 83959-induced CPA was abolished by pre-treatment by the D1-D2 heteromer selective disrupting peptide, TAT-D1, but not the control TAT-Sc peptide. (C) Inactivation of D1-D2 heteromer by TAT-D1 resulted in conditioned place preference (CPP) as the rats spent significantly more time in the drug paired chamber, not observed with the control TAT-Sc. (D) Representative western blots (inset) and histogram showing the amount of D1R co-immunoprecipitated with D2R from the NAc of rats treated with saline or SKF 83959. Pretreatment with TAT-D1 led to decreased co-immunoprecipitated receptors. An aliquot of each sample was used as a control for WB (input control). (E,F) The CPA induced by D1-D2 heteromer stimulation was abolished by Cdk5 inhibitor roscovitine pre-treatment (200 nmol, i.c.v, E ) or intra-accumbal injection (30 nmol, F ). (G) Representative western blot and histogram showing the density of Thr75-DARPP-32 phosphorylation (pT75) relative to GAPDH (as loading control). Data represent means ± SEM of n = 8–10 rats/group. ( * p

    Techniques Used: Conditioned Place Preference, Western Blot, Immunoprecipitation, Injection

    Evidence for the existence of dopamine D1-D2 receptor heteromer in rat NAc. (A) Proximity ligation assay (PLA) was used to visualize and detect D1R and D2R close proximity. (A1) A scheme depicts the PLA probes used in the present study. (A2–A4) Representative images of PLA signals (red dots) in neurons (nuclei stained by DAPI) in rat caudate putamen (CPu), nucleus accumbens core (NAc-core) and shell (NAc-shell) subregions. (A5) Graph representing the percent of neurons with a positive PLA signal. (A6) Representative image of PLA signals in neurons (nuclei stained by DAPI) in rat NAc-core using the second set of antibodies. (B) Representative images of immunohistochemistry using D1R antibody (D1R-Ab) or D2R antibody (D2R-Ab) directly conjugated to Alexa-488 or Alexa-568, respectively, in the NAc-shell. Direct confocal FRET analysis was performed, reflected by FRET efficiency (FRET E) and the distance between the dipoles, less than 10 nm (100 Å). (C) A representative close-up of a single MSN cell body from NAc showing D1R-D2R colocalization (left), D1-D2 heteromer FRET efficiency (center) and relative distance between receptors (right). (D,E) Histograms showing FRET E ratios (D) and distance (E) obtained from MSN cell bodies from NAc ( n = 24). Bars are 10 μm.
    Figure Legend Snippet: Evidence for the existence of dopamine D1-D2 receptor heteromer in rat NAc. (A) Proximity ligation assay (PLA) was used to visualize and detect D1R and D2R close proximity. (A1) A scheme depicts the PLA probes used in the present study. (A2–A4) Representative images of PLA signals (red dots) in neurons (nuclei stained by DAPI) in rat caudate putamen (CPu), nucleus accumbens core (NAc-core) and shell (NAc-shell) subregions. (A5) Graph representing the percent of neurons with a positive PLA signal. (A6) Representative image of PLA signals in neurons (nuclei stained by DAPI) in rat NAc-core using the second set of antibodies. (B) Representative images of immunohistochemistry using D1R antibody (D1R-Ab) or D2R antibody (D2R-Ab) directly conjugated to Alexa-488 or Alexa-568, respectively, in the NAc-shell. Direct confocal FRET analysis was performed, reflected by FRET efficiency (FRET E) and the distance between the dipoles, less than 10 nm (100 Å). (C) A representative close-up of a single MSN cell body from NAc showing D1R-D2R colocalization (left), D1-D2 heteromer FRET efficiency (center) and relative distance between receptors (right). (D,E) Histograms showing FRET E ratios (D) and distance (E) obtained from MSN cell bodies from NAc ( n = 24). Bars are 10 μm.

    Techniques Used: Proximity Ligation Assay, Staining, Immunohistochemistry

    2) Product Images from "Central Thalamic Deep-Brain Stimulation Alters Striatal-Thalamic Connectivity in Cognitive Neural Behavior"

    Article Title: Central Thalamic Deep-Brain Stimulation Alters Striatal-Thalamic Connectivity in Cognitive Neural Behavior

    Journal: Frontiers in Neural Circuits

    doi: 10.3389/fncir.2015.00087

    Western blot protein analysis of cells excised individually from striatal and hippocampal tissues. (A) The SDS-PAGE blots show the expression of dopamine D2 receptor (Drd2) and α4-nicotinic acetylcholine receptor (α4-nAChR) in the striatum (top) and hippocampus (bottom). (B) Results from the quantitative analysis of Drd2 and α4-nAChR expression (mean ± SEM; expressed as ratio to GAPDH) in the striatum (top) and hippocampus (bottom). Striatal and Drd2 and α4-nAChR protein expressions were significantly increased relative to sham control group. Significant increases in Drd2 and α4-nAChR protein expressions in the hippocampus also were found. * , ** , and *** indicate significant protein expression with P
    Figure Legend Snippet: Western blot protein analysis of cells excised individually from striatal and hippocampal tissues. (A) The SDS-PAGE blots show the expression of dopamine D2 receptor (Drd2) and α4-nicotinic acetylcholine receptor (α4-nAChR) in the striatum (top) and hippocampus (bottom). (B) Results from the quantitative analysis of Drd2 and α4-nAChR expression (mean ± SEM; expressed as ratio to GAPDH) in the striatum (top) and hippocampus (bottom). Striatal and Drd2 and α4-nAChR protein expressions were significantly increased relative to sham control group. Significant increases in Drd2 and α4-nAChR protein expressions in the hippocampus also were found. * , ** , and *** indicate significant protein expression with P

    Techniques Used: Western Blot, SDS Page, Expressing

    3) Product Images from "A2A-D2 Heteromers on Striatal Astrocytes: Biochemical and Biophysical Evidence"

    Article Title: A2A-D2 Heteromers on Striatal Astrocytes: Biochemical and Biophysical Evidence

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms20102457

    A2A-D2 heterodimers on rat striatal astrocytes: proximity ligation assay. Detection of in situ PLA A2A-D2 heteroreceptor complexes was carried out with primary antibodies (rabbit polyclonal anti-A2AR, mouse monoclonal anti-D2R and goat polyclonal anti-GFAP) in rat striatal slices. ( A ) The merge of the maximum intensity projections of a representative field (240x240 µm; z 10 µm) is shown; GFAP (red), DAPI (blue), PLA for A2A-D2 heteroreceptor complexes appears as yellow clusters. The boxed region (i) is shown at a higher magnification in ( B ). ( C ) Merged confocal images of a single z stack of the boxed region (ii) at a higher magnification: double immunolabeling for GFAP (red) and PLA A2A-D2 heteroreceptor complexes (green). ( D ) Colocalized map of the boxed region (ii); the colocalization analysis plugins—colocalization threshold with GFAP as ROI—was used to create the colocalized map; ImageJ Fiji software. A complete lack of stain for PLA A2A-D2 heteroreceptor complexes was obtained in the negative control experiments, performed avoiding the conjugation of a primary antibody with the Duolink Probes. In the figure the merges of the maximum intensity projections of two representative fields are shown: PLA for A2A-D2. heteroreceptor complexes without the primary anti-A2A antibody ( E ) or the primary anti-D2 antibody ( F ). Scale bar 25 µm or 10 µm
    Figure Legend Snippet: A2A-D2 heterodimers on rat striatal astrocytes: proximity ligation assay. Detection of in situ PLA A2A-D2 heteroreceptor complexes was carried out with primary antibodies (rabbit polyclonal anti-A2AR, mouse monoclonal anti-D2R and goat polyclonal anti-GFAP) in rat striatal slices. ( A ) The merge of the maximum intensity projections of a representative field (240x240 µm; z 10 µm) is shown; GFAP (red), DAPI (blue), PLA for A2A-D2 heteroreceptor complexes appears as yellow clusters. The boxed region (i) is shown at a higher magnification in ( B ). ( C ) Merged confocal images of a single z stack of the boxed region (ii) at a higher magnification: double immunolabeling for GFAP (red) and PLA A2A-D2 heteroreceptor complexes (green). ( D ) Colocalized map of the boxed region (ii); the colocalization analysis plugins—colocalization threshold with GFAP as ROI—was used to create the colocalized map; ImageJ Fiji software. A complete lack of stain for PLA A2A-D2 heteroreceptor complexes was obtained in the negative control experiments, performed avoiding the conjugation of a primary antibody with the Duolink Probes. In the figure the merges of the maximum intensity projections of two representative fields are shown: PLA for A2A-D2. heteroreceptor complexes without the primary anti-A2A antibody ( E ) or the primary anti-D2 antibody ( F ). Scale bar 25 µm or 10 µm

    Techniques Used: Proximity Ligation Assay, In Situ, Immunolabeling, Software, Staining, Negative Control, Conjugation Assay

    4) Product Images from "Activation of Dopamine D1-D2 Receptor Complex Attenuates Cocaine Reward and Reinstatement of Cocaine-Seeking through Inhibition of DARPP-32, ERK, and ΔFosB"

    Article Title: Activation of Dopamine D1-D2 Receptor Complex Attenuates Cocaine Reward and Reinstatement of Cocaine-Seeking through Inhibition of DARPP-32, ERK, and ΔFosB

    Journal: Frontiers in Pharmacology

    doi: 10.3389/fphar.2017.00924

    The effects of D1-D2 heteromer stimulation and inactivation on basal conditioned place preference. (A) Vehicle-conditioned rats did not exhibit a preference toward a particular chamber. D1-D2 heteromer stimulation by SKF 83959 (1.5 mg/kg, s.c.) induced conditioned place aversion (CPA) as the animals spent significantly less time in the drug paired chamber. (B) SKF 83959-induced CPA was abolished by pre-treatment by the D1-D2 heteromer selective disrupting peptide, TAT-D1, but not the control TAT-Sc peptide. (C) Inactivation of D1-D2 heteromer by TAT-D1 resulted in conditioned place preference (CPP) as the rats spent significantly more time in the drug paired chamber, not observed with the control TAT-Sc. (D) Representative western blots (inset) and histogram showing the amount of D1R co-immunoprecipitated with D2R from the NAc of rats treated with saline or SKF 83959. Pretreatment with TAT-D1 led to decreased co-immunoprecipitated receptors. An aliquot of each sample was used as a control for WB (input control). (E,F) The CPA induced by D1-D2 heteromer stimulation was abolished by Cdk5 inhibitor roscovitine pre-treatment (200 nmol, i.c.v, E ) or intra-accumbal injection (30 nmol, F ). (G) Representative western blot and histogram showing the density of Thr75-DARPP-32 phosphorylation (pT75) relative to GAPDH (as loading control). Data represent means ± SEM of n = 8–10 rats/group. ( * p
    Figure Legend Snippet: The effects of D1-D2 heteromer stimulation and inactivation on basal conditioned place preference. (A) Vehicle-conditioned rats did not exhibit a preference toward a particular chamber. D1-D2 heteromer stimulation by SKF 83959 (1.5 mg/kg, s.c.) induced conditioned place aversion (CPA) as the animals spent significantly less time in the drug paired chamber. (B) SKF 83959-induced CPA was abolished by pre-treatment by the D1-D2 heteromer selective disrupting peptide, TAT-D1, but not the control TAT-Sc peptide. (C) Inactivation of D1-D2 heteromer by TAT-D1 resulted in conditioned place preference (CPP) as the rats spent significantly more time in the drug paired chamber, not observed with the control TAT-Sc. (D) Representative western blots (inset) and histogram showing the amount of D1R co-immunoprecipitated with D2R from the NAc of rats treated with saline or SKF 83959. Pretreatment with TAT-D1 led to decreased co-immunoprecipitated receptors. An aliquot of each sample was used as a control for WB (input control). (E,F) The CPA induced by D1-D2 heteromer stimulation was abolished by Cdk5 inhibitor roscovitine pre-treatment (200 nmol, i.c.v, E ) or intra-accumbal injection (30 nmol, F ). (G) Representative western blot and histogram showing the density of Thr75-DARPP-32 phosphorylation (pT75) relative to GAPDH (as loading control). Data represent means ± SEM of n = 8–10 rats/group. ( * p

    Techniques Used: Conditioned Place Preference, Western Blot, Immunoprecipitation, Injection

    Evidence for the existence of dopamine D1-D2 receptor heteromer in rat NAc. (A) Proximity ligation assay (PLA) was used to visualize and detect D1R and D2R close proximity. (A1) A scheme depicts the PLA probes used in the present study. (A2–A4) Representative images of PLA signals (red dots) in neurons (nuclei stained by DAPI) in rat caudate putamen (CPu), nucleus accumbens core (NAc-core) and shell (NAc-shell) subregions. (A5) Graph representing the percent of neurons with a positive PLA signal. (A6) Representative image of PLA signals in neurons (nuclei stained by DAPI) in rat NAc-core using the second set of antibodies. (B) Representative images of immunohistochemistry using D1R antibody (D1R-Ab) or D2R antibody (D2R-Ab) directly conjugated to Alexa-488 or Alexa-568, respectively, in the NAc-shell. Direct confocal FRET analysis was performed, reflected by FRET efficiency (FRET E) and the distance between the dipoles, less than 10 nm (100 Å). (C) A representative close-up of a single MSN cell body from NAc showing D1R-D2R colocalization (left), D1-D2 heteromer FRET efficiency (center) and relative distance between receptors (right). (D,E) Histograms showing FRET E ratios (D) and distance (E) obtained from MSN cell bodies from NAc ( n = 24). Bars are 10 μm.
    Figure Legend Snippet: Evidence for the existence of dopamine D1-D2 receptor heteromer in rat NAc. (A) Proximity ligation assay (PLA) was used to visualize and detect D1R and D2R close proximity. (A1) A scheme depicts the PLA probes used in the present study. (A2–A4) Representative images of PLA signals (red dots) in neurons (nuclei stained by DAPI) in rat caudate putamen (CPu), nucleus accumbens core (NAc-core) and shell (NAc-shell) subregions. (A5) Graph representing the percent of neurons with a positive PLA signal. (A6) Representative image of PLA signals in neurons (nuclei stained by DAPI) in rat NAc-core using the second set of antibodies. (B) Representative images of immunohistochemistry using D1R antibody (D1R-Ab) or D2R antibody (D2R-Ab) directly conjugated to Alexa-488 or Alexa-568, respectively, in the NAc-shell. Direct confocal FRET analysis was performed, reflected by FRET efficiency (FRET E) and the distance between the dipoles, less than 10 nm (100 Å). (C) A representative close-up of a single MSN cell body from NAc showing D1R-D2R colocalization (left), D1-D2 heteromer FRET efficiency (center) and relative distance between receptors (right). (D,E) Histograms showing FRET E ratios (D) and distance (E) obtained from MSN cell bodies from NAc ( n = 24). Bars are 10 μm.

    Techniques Used: Proximity Ligation Assay, Staining, Immunohistochemistry

    5) Product Images from "FFAR1/GPR40 Contributes to the Regulation of Striatal Monoamine Releases and Facilitation of Cocaine-Induced Locomotor Activity in Mice"

    Article Title: FFAR1/GPR40 Contributes to the Regulation of Striatal Monoamine Releases and Facilitation of Cocaine-Induced Locomotor Activity in Mice

    Journal: Frontiers in Pharmacology

    doi: 10.3389/fphar.2021.699026

    Quantitative PCR and western blot analyses of DA and 5-HT transporters, and dopamine D1 and D2 receptors in the striatum. qPCR (A–D) and western blot (E–I) analyses showing that no significant differences in the expression levels of DA transporter (DAT: A,E,I ), 5-HT transporter (5HTT: B,F,I ), D1 receptor (D1R: C,G,I ), and D2 receptor (D2R: D,H,I ) were observed between FFAR1+/+ and −/− mice. n = 6 (A) and 5 (B-D) each. In (I) , n = 4 (+/+) and 6 (−/−) for DAT, n = 8.
    Figure Legend Snippet: Quantitative PCR and western blot analyses of DA and 5-HT transporters, and dopamine D1 and D2 receptors in the striatum. qPCR (A–D) and western blot (E–I) analyses showing that no significant differences in the expression levels of DA transporter (DAT: A,E,I ), 5-HT transporter (5HTT: B,F,I ), D1 receptor (D1R: C,G,I ), and D2 receptor (D2R: D,H,I ) were observed between FFAR1+/+ and −/− mice. n = 6 (A) and 5 (B-D) each. In (I) , n = 4 (+/+) and 6 (−/−) for DAT, n = 8.

    Techniques Used: Real-time Polymerase Chain Reaction, Western Blot, Expressing, Mouse Assay

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    Alomone Labs dopamine d2r for co ip
    The effects of D1-D2 heteromer stimulation and inactivation on basal conditioned place preference. (A) Vehicle-conditioned rats did not exhibit a preference toward a particular chamber. D1-D2 heteromer stimulation by SKF 83959 (1.5 mg/kg, s.c.) induced conditioned place aversion (CPA) as the animals spent significantly less time in the drug paired chamber. (B) SKF 83959-induced CPA was abolished by pre-treatment by the D1-D2 heteromer selective disrupting peptide, TAT-D1, but not the control TAT-Sc peptide. (C) Inactivation of D1-D2 heteromer by TAT-D1 resulted in conditioned place preference (CPP) as the rats spent significantly more time in the drug paired chamber, not observed with the control TAT-Sc. (D) Representative western blots (inset) and histogram showing the amount of D1R co-immunoprecipitated with <t>D2R</t> from the NAc of rats treated with saline or SKF 83959. Pretreatment with TAT-D1 led to decreased co-immunoprecipitated receptors. An aliquot of each sample was used as a control for WB (input control). (E,F) The CPA induced by D1-D2 heteromer stimulation was abolished by Cdk5 inhibitor roscovitine pre-treatment (200 nmol, i.c.v, E ) or intra-accumbal injection (30 nmol, F ). (G) Representative western blot and histogram showing the density of Thr75-DARPP-32 phosphorylation (pT75) relative to GAPDH (as loading control). Data represent means ± SEM of n = 8–10 rats/group. ( * p
    Dopamine D2r For Co Ip, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dopamine d2r for co ip/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    dopamine d2r for co ip - by Bioz Stars, 2022-07
    93/100 stars
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    The effects of D1-D2 heteromer stimulation and inactivation on basal conditioned place preference. (A) Vehicle-conditioned rats did not exhibit a preference toward a particular chamber. D1-D2 heteromer stimulation by SKF 83959 (1.5 mg/kg, s.c.) induced conditioned place aversion (CPA) as the animals spent significantly less time in the drug paired chamber. (B) SKF 83959-induced CPA was abolished by pre-treatment by the D1-D2 heteromer selective disrupting peptide, TAT-D1, but not the control TAT-Sc peptide. (C) Inactivation of D1-D2 heteromer by TAT-D1 resulted in conditioned place preference (CPP) as the rats spent significantly more time in the drug paired chamber, not observed with the control TAT-Sc. (D) Representative western blots (inset) and histogram showing the amount of D1R co-immunoprecipitated with D2R from the NAc of rats treated with saline or SKF 83959. Pretreatment with TAT-D1 led to decreased co-immunoprecipitated receptors. An aliquot of each sample was used as a control for WB (input control). (E,F) The CPA induced by D1-D2 heteromer stimulation was abolished by Cdk5 inhibitor roscovitine pre-treatment (200 nmol, i.c.v, E ) or intra-accumbal injection (30 nmol, F ). (G) Representative western blot and histogram showing the density of Thr75-DARPP-32 phosphorylation (pT75) relative to GAPDH (as loading control). Data represent means ± SEM of n = 8–10 rats/group. ( * p

    Journal: Frontiers in Pharmacology

    Article Title: Activation of Dopamine D1-D2 Receptor Complex Attenuates Cocaine Reward and Reinstatement of Cocaine-Seeking through Inhibition of DARPP-32, ERK, and ΔFosB

    doi: 10.3389/fphar.2017.00924

    Figure Lengend Snippet: The effects of D1-D2 heteromer stimulation and inactivation on basal conditioned place preference. (A) Vehicle-conditioned rats did not exhibit a preference toward a particular chamber. D1-D2 heteromer stimulation by SKF 83959 (1.5 mg/kg, s.c.) induced conditioned place aversion (CPA) as the animals spent significantly less time in the drug paired chamber. (B) SKF 83959-induced CPA was abolished by pre-treatment by the D1-D2 heteromer selective disrupting peptide, TAT-D1, but not the control TAT-Sc peptide. (C) Inactivation of D1-D2 heteromer by TAT-D1 resulted in conditioned place preference (CPP) as the rats spent significantly more time in the drug paired chamber, not observed with the control TAT-Sc. (D) Representative western blots (inset) and histogram showing the amount of D1R co-immunoprecipitated with D2R from the NAc of rats treated with saline or SKF 83959. Pretreatment with TAT-D1 led to decreased co-immunoprecipitated receptors. An aliquot of each sample was used as a control for WB (input control). (E,F) The CPA induced by D1-D2 heteromer stimulation was abolished by Cdk5 inhibitor roscovitine pre-treatment (200 nmol, i.c.v, E ) or intra-accumbal injection (30 nmol, F ). (G) Representative western blot and histogram showing the density of Thr75-DARPP-32 phosphorylation (pT75) relative to GAPDH (as loading control). Data represent means ± SEM of n = 8–10 rats/group. ( * p

    Article Snippet: AntibodiesThe primary antibodies used in the present study were raised against: Dopamine D1R (Sigma, D2944, rat); dopamine D2R for IHC and PLA (1st set) (Millipore; AB5084P; rabbit), dopamine D2R for co-IP (Alomone; Rabbit), dopamine D2R for second set PLA (Millipore, ABN 462), phospho-Thr75-DARPP-32 (Cell signaling, 2301s; rabbit), phospho-Thr34-DARPP-32 (Cell Signaling, 2304; rabbit), enkephalin (Millipore, MAB350, mouse), phospho-ERK1/2 (Sigma, E7028; rabbit), ΔFosB (Cell signaling, 14695S, rabbit), GAPDH (Millipore, rabbit).

    Techniques: Conditioned Place Preference, Western Blot, Immunoprecipitation, Injection

    Evidence for the existence of dopamine D1-D2 receptor heteromer in rat NAc. (A) Proximity ligation assay (PLA) was used to visualize and detect D1R and D2R close proximity. (A1) A scheme depicts the PLA probes used in the present study. (A2–A4) Representative images of PLA signals (red dots) in neurons (nuclei stained by DAPI) in rat caudate putamen (CPu), nucleus accumbens core (NAc-core) and shell (NAc-shell) subregions. (A5) Graph representing the percent of neurons with a positive PLA signal. (A6) Representative image of PLA signals in neurons (nuclei stained by DAPI) in rat NAc-core using the second set of antibodies. (B) Representative images of immunohistochemistry using D1R antibody (D1R-Ab) or D2R antibody (D2R-Ab) directly conjugated to Alexa-488 or Alexa-568, respectively, in the NAc-shell. Direct confocal FRET analysis was performed, reflected by FRET efficiency (FRET E) and the distance between the dipoles, less than 10 nm (100 Å). (C) A representative close-up of a single MSN cell body from NAc showing D1R-D2R colocalization (left), D1-D2 heteromer FRET efficiency (center) and relative distance between receptors (right). (D,E) Histograms showing FRET E ratios (D) and distance (E) obtained from MSN cell bodies from NAc ( n = 24). Bars are 10 μm.

    Journal: Frontiers in Pharmacology

    Article Title: Activation of Dopamine D1-D2 Receptor Complex Attenuates Cocaine Reward and Reinstatement of Cocaine-Seeking through Inhibition of DARPP-32, ERK, and ΔFosB

    doi: 10.3389/fphar.2017.00924

    Figure Lengend Snippet: Evidence for the existence of dopamine D1-D2 receptor heteromer in rat NAc. (A) Proximity ligation assay (PLA) was used to visualize and detect D1R and D2R close proximity. (A1) A scheme depicts the PLA probes used in the present study. (A2–A4) Representative images of PLA signals (red dots) in neurons (nuclei stained by DAPI) in rat caudate putamen (CPu), nucleus accumbens core (NAc-core) and shell (NAc-shell) subregions. (A5) Graph representing the percent of neurons with a positive PLA signal. (A6) Representative image of PLA signals in neurons (nuclei stained by DAPI) in rat NAc-core using the second set of antibodies. (B) Representative images of immunohistochemistry using D1R antibody (D1R-Ab) or D2R antibody (D2R-Ab) directly conjugated to Alexa-488 or Alexa-568, respectively, in the NAc-shell. Direct confocal FRET analysis was performed, reflected by FRET efficiency (FRET E) and the distance between the dipoles, less than 10 nm (100 Å). (C) A representative close-up of a single MSN cell body from NAc showing D1R-D2R colocalization (left), D1-D2 heteromer FRET efficiency (center) and relative distance between receptors (right). (D,E) Histograms showing FRET E ratios (D) and distance (E) obtained from MSN cell bodies from NAc ( n = 24). Bars are 10 μm.

    Article Snippet: AntibodiesThe primary antibodies used in the present study were raised against: Dopamine D1R (Sigma, D2944, rat); dopamine D2R for IHC and PLA (1st set) (Millipore; AB5084P; rabbit), dopamine D2R for co-IP (Alomone; Rabbit), dopamine D2R for second set PLA (Millipore, ABN 462), phospho-Thr75-DARPP-32 (Cell signaling, 2301s; rabbit), phospho-Thr34-DARPP-32 (Cell Signaling, 2304; rabbit), enkephalin (Millipore, MAB350, mouse), phospho-ERK1/2 (Sigma, E7028; rabbit), ΔFosB (Cell signaling, 14695S, rabbit), GAPDH (Millipore, rabbit).

    Techniques: Proximity Ligation Assay, Staining, Immunohistochemistry

    Western blot protein analysis of cells excised individually from striatal and hippocampal tissues. (A) The SDS-PAGE blots show the expression of dopamine D2 receptor (Drd2) and α4-nicotinic acetylcholine receptor (α4-nAChR) in the striatum (top) and hippocampus (bottom). (B) Results from the quantitative analysis of Drd2 and α4-nAChR expression (mean ± SEM; expressed as ratio to GAPDH) in the striatum (top) and hippocampus (bottom). Striatal and Drd2 and α4-nAChR protein expressions were significantly increased relative to sham control group. Significant increases in Drd2 and α4-nAChR protein expressions in the hippocampus also were found. * , ** , and *** indicate significant protein expression with P

    Journal: Frontiers in Neural Circuits

    Article Title: Central Thalamic Deep-Brain Stimulation Alters Striatal-Thalamic Connectivity in Cognitive Neural Behavior

    doi: 10.3389/fncir.2015.00087

    Figure Lengend Snippet: Western blot protein analysis of cells excised individually from striatal and hippocampal tissues. (A) The SDS-PAGE blots show the expression of dopamine D2 receptor (Drd2) and α4-nicotinic acetylcholine receptor (α4-nAChR) in the striatum (top) and hippocampus (bottom). (B) Results from the quantitative analysis of Drd2 and α4-nAChR expression (mean ± SEM; expressed as ratio to GAPDH) in the striatum (top) and hippocampus (bottom). Striatal and Drd2 and α4-nAChR protein expressions were significantly increased relative to sham control group. Significant increases in Drd2 and α4-nAChR protein expressions in the hippocampus also were found. * , ** , and *** indicate significant protein expression with P

    Article Snippet: The membranes were hybridized with anti-dopamine D2 receptor (Drd2; 1:1000; ADR-002-50UL, Alomone Labs, Jerusalem, Israel) or anti-nicotinic acetylcholine α4 receptor (α4-nAChR; 1:1000; ANC-004-50UL, Alomone Labs, Jerusalem, Israel) antibodies.

    Techniques: Western Blot, SDS Page, Expressing

    A2A-D2 heterodimers on rat striatal astrocytes: proximity ligation assay. Detection of in situ PLA A2A-D2 heteroreceptor complexes was carried out with primary antibodies (rabbit polyclonal anti-A2AR, mouse monoclonal anti-D2R and goat polyclonal anti-GFAP) in rat striatal slices. ( A ) The merge of the maximum intensity projections of a representative field (240x240 µm; z 10 µm) is shown; GFAP (red), DAPI (blue), PLA for A2A-D2 heteroreceptor complexes appears as yellow clusters. The boxed region (i) is shown at a higher magnification in ( B ). ( C ) Merged confocal images of a single z stack of the boxed region (ii) at a higher magnification: double immunolabeling for GFAP (red) and PLA A2A-D2 heteroreceptor complexes (green). ( D ) Colocalized map of the boxed region (ii); the colocalization analysis plugins—colocalization threshold with GFAP as ROI—was used to create the colocalized map; ImageJ Fiji software. A complete lack of stain for PLA A2A-D2 heteroreceptor complexes was obtained in the negative control experiments, performed avoiding the conjugation of a primary antibody with the Duolink Probes. In the figure the merges of the maximum intensity projections of two representative fields are shown: PLA for A2A-D2. heteroreceptor complexes without the primary anti-A2A antibody ( E ) or the primary anti-D2 antibody ( F ). Scale bar 25 µm or 10 µm

    Journal: International Journal of Molecular Sciences

    Article Title: A2A-D2 Heteromers on Striatal Astrocytes: Biochemical and Biophysical Evidence

    doi: 10.3390/ijms20102457

    Figure Lengend Snippet: A2A-D2 heterodimers on rat striatal astrocytes: proximity ligation assay. Detection of in situ PLA A2A-D2 heteroreceptor complexes was carried out with primary antibodies (rabbit polyclonal anti-A2AR, mouse monoclonal anti-D2R and goat polyclonal anti-GFAP) in rat striatal slices. ( A ) The merge of the maximum intensity projections of a representative field (240x240 µm; z 10 µm) is shown; GFAP (red), DAPI (blue), PLA for A2A-D2 heteroreceptor complexes appears as yellow clusters. The boxed region (i) is shown at a higher magnification in ( B ). ( C ) Merged confocal images of a single z stack of the boxed region (ii) at a higher magnification: double immunolabeling for GFAP (red) and PLA A2A-D2 heteroreceptor complexes (green). ( D ) Colocalized map of the boxed region (ii); the colocalization analysis plugins—colocalization threshold with GFAP as ROI—was used to create the colocalized map; ImageJ Fiji software. A complete lack of stain for PLA A2A-D2 heteroreceptor complexes was obtained in the negative control experiments, performed avoiding the conjugation of a primary antibody with the Duolink Probes. In the figure the merges of the maximum intensity projections of two representative fields are shown: PLA for A2A-D2. heteroreceptor complexes without the primary anti-A2A antibody ( E ) or the primary anti-D2 antibody ( F ). Scale bar 25 µm or 10 µm

    Article Snippet: In situ PLA was performed on 10µm rat striatal slices using the primary antibodies (mouse anti-A2A (1:200, Merck Millipore Corporation, 05-717, Burlington, MA, USA); rabbit anti-D2R (1:200, Alomone Labs, ADR-002, Jerusalem, Israel); goat anti-GFAP (1:500, Santa Cruz Biotechnoloy Inc, Dallas, TX, USA, sc-6170), the Duolink in situ PLA detection kit (DUO92014, Sigma-Aldrich, St Louis, MO, USA) and Alexa Fluor 546-conjugated donkey anti-goat (1:500; Molecular Probes, Eugene, OR, USA).

    Techniques: Proximity Ligation Assay, In Situ, Immunolabeling, Software, Staining, Negative Control, Conjugation Assay