Journal: Molecular neurobiology

Article Title: The mGlu 5 Receptor Protomer-Mediated Dopamine D 2 Receptor Trans-Inhibition Is Dependent on the Adenosine A 2A Receptor Protomer: Implications for Parkinson's Disease.

doi: 10.1007/s12035-022-02946-9

Figure Lengend Snippet: Fig. 2 In situ PLA assessment of D2R-mGluR5 heteromer formation in the absence (A) or presence (B) of A2AR (see “Methods”). The in situ PLA-positive D2R-mGluR5 heteroreceptor complexes were shown as red blobs (arrows) and nuclei in blue (DAPI staining). A negative in situ PLA control (C) was included by incubating the cells in the absence of the primary anti-D2R antibody. D Quantification of D2R-mGluR5 complexes. The number of PLA blobs (red clusters) per positive cell (n = 4 × 50 cells) was assessed as described in Methods. Results were expressed as mean ± SEM (n = 4 independent experi- ments). ****p < 0.0001 and **p < 0.01, Student’s t-test

Article Snippet: The A2AR agonist 4-[2-[[6-Amino-9-(N-ethyl-β-Dribofuranuronamidosyl)-9H-purin-2-yl]amino]ethyl]benzenepropanoic acid hydrochloride (CGS-21680), the selective A2AR antagonist 4-(2-[7-Amino-2-(2-furyl)[1,2,4] triazolo[2,3-a][1,3,5]triazin-5-ylamino]ethyl)phenol (ZM241385), the mGluR5 agonist (RS)-2-Chloro-5-hydroxyphenylglycine sodium salt (CHPG), the mGluR5 antagonist 2-Methyl-6-(phenylethynyl)pyridine hydrochloride (MPEP) and the D2R antagonist 4-[4-(4-Chlorophenyl)-4-hydroxy-1piperidinyl]-1-(4-fluorophenyl)-1-butanone hydrochloride (haloperidol) were purchased from Tocris Bioscience (UK), and the mGluR5 negative allosteric modulator 2-[(3-Fluorophenyl)ethynyl]-4,6-dimethyl-3-pyridinamine hydrochloride (raseglurant) was purchased from Hello Bio (Republic of Ireland).

Techniques: In Situ, Staining, Control

Journal: Molecular neurobiology

Article Title: The mGlu 5 Receptor Protomer-Mediated Dopamine D 2 Receptor Trans-Inhibition Is Dependent on the Adenosine A 2A Receptor Protomer: Implications for Parkinson's Disease.

doi: 10.1007/s12035-022-02946-9

Figure Lengend Snippet: Fig. 3 Assessment of D2R-mGluR5 heteromer formation in mouse dorsal striatum by in situ PLA. Photomicrographs showing PLA recognition of D2R-mGluR5 heteromers in the dorsal striatum of wild type (A), A2AR–/– (B) and D2R–/– (C) mice. The in situ PLA- positive D2R-mGluR5 heteroreceptor complexes are shown as red blobs (arrows) and nuclei in blue (DAPI staining). D Quantification of D2R-mGluR5 complexes showing a highly significant reduction of D2R-mGluR5-positive red blobs in the absence of A2AR–/– or D2R–/.–. The number of PLA blobs (red clusters) per nucleus was assessed as described in Methods. Results were expressed as mean ± SEM (n = 5 animals). ***p < 0.001, one-way ANOVA followed by Dunnett’s post hoc test when compared with wild-type animals

Article Snippet: The A2AR agonist 4-[2-[[6-Amino-9-(N-ethyl-β-Dribofuranuronamidosyl)-9H-purin-2-yl]amino]ethyl]benzenepropanoic acid hydrochloride (CGS-21680), the selective A2AR antagonist 4-(2-[7-Amino-2-(2-furyl)[1,2,4] triazolo[2,3-a][1,3,5]triazin-5-ylamino]ethyl)phenol (ZM241385), the mGluR5 agonist (RS)-2-Chloro-5-hydroxyphenylglycine sodium salt (CHPG), the mGluR5 antagonist 2-Methyl-6-(phenylethynyl)pyridine hydrochloride (MPEP) and the D2R antagonist 4-[4-(4-Chlorophenyl)-4-hydroxy-1piperidinyl]-1-(4-fluorophenyl)-1-butanone hydrochloride (haloperidol) were purchased from Tocris Bioscience (UK), and the mGluR5 negative allosteric modulator 2-[(3-Fluorophenyl)ethynyl]-4,6-dimethyl-3-pyridinamine hydrochloride (raseglurant) was purchased from Hello Bio (Republic of Ireland).

Techniques: In Situ, Staining