h ro15 4513  (Revvity)

Journal: bioRxiv

Article Title: Erbb4 deletion from fast-spiking interneurons causes psychosis-relevant neuroimaging phenotypes

doi: 10.1101/2022.03.07.483347

Figure Lengend Snippet: Representative hippocampal regions of interests. (A) Regions of interests sampled for [ 3 H]Ro15-4513 and [ 3 H]flumazenil. (B) Regions of interests sampled for [ 3 H]UCB-J. dHip CA1: dorsal hippocampus CA1; mHip CA3: middle hippocampus CA3; mHip CA1/2: middle hippocampus CA1/CA2; vHip CA3: ventral hippocampus CA3; dHip DG: dorsal hippocampus dentate gyrus; mHip DG: middle hippocampus dentate gyrus.

Article Snippet: To quantify density of α5GABA A R – sections were incubated for 60 minutes at room temperature in 2nM [ 3 H]Ro15-4513 (Perkin Elmer, NET925250UC), or in 2nM [ 3 H]Ro15-4513 with 10 µM bretazenil (Sigma, B6434) for nonspecific binding.

Techniques:

Journal: bioRxiv

Article Title: Erbb4 deletion from fast-spiking interneurons causes psychosis-relevant neuroimaging phenotypes

doi: 10.1101/2022.03.07.483347

Figure Lengend Snippet: [ 3 H]UCB-J, but not [ 3 H]Ro15-4513 or [ 3 H]flumazenil binding in Lhx6-Cre;Erbb4 F/F mice is decreased across the hippocampus. (A) [ 3 H]UCB-J showed a significant decrease in synaptic density in Lhx6-Cre;Erbb4 F/F mice (n=11) compared to control animals (n=9) across all hippocampal ROIs (mixed effects analysis; main effect of genotype: F (1,18) =7.27, p =0.02, ). (B) [ 3 H]Ro15-4513 (n=10-11/group) and (C) [ 3 H]flumazenil (n=9-10/group) binding did not differ by genotype. dHip CA1: dorsal hippocampus CA1; mHip CA3: middle hippocampus CA3; mHip CA1/2: middle hippocampus CA1/CA2; vHip CA3: ventral hippocampus CA3; dHip DG: dorsal hippocampus dentate gyrus; mHip DG: dentate gyrus.

Article Snippet: To quantify density of α5GABA A R – sections were incubated for 60 minutes at room temperature in 2nM [ 3 H]Ro15-4513 (Perkin Elmer, NET925250UC), or in 2nM [ 3 H]Ro15-4513 with 10 µM bretazenil (Sigma, B6434) for nonspecific binding.

Techniques: Binding Assay

Journal: The Journal of Neuroscience

Article Title: Loss of the Major GABA A Receptor Subtype in the Brain Is Not Lethal in Mice

doi: 10.1523/JNEUROSCI.21-10-03409.2001

Figure Lengend Snippet: Amount (fmol/mg protein) of GABA A receptors and benzodiazepine binding sites in WT, α1−/−, and β2−/− mice determined by nonlinear regression analysis of saturation data sets

Article Snippet: Radioligand binding assays with [ 3 H]Ro15–1788 (87 Ci/mmol; NEN Life Sciences), [ 3 H]Ro15–4513 (21.7 Ci/mmol; NEN Life Sciences) in the presence or absence of 10 μ m diazepam and [ 3 H]muscimol (19.1 Ci/mmol; NEN Life Sciences) were performed on membrane preparations as previously described ( Quirk et al., 1994 ; Sur et al., 1998 , 1999a ).

Techniques: Binding Assay

Journal: The Journal of Neuroscience

Article Title: Loss of the Major GABA A Receptor Subtype in the Brain Is Not Lethal in Mice

doi: 10.1523/JNEUROSCI.21-10-03409.2001

Figure Lengend Snippet: Pharmacological characterization of GABA A receptors in WT, α1 (−/−), and β2 (−/−) mice

Article Snippet: Radioligand binding assays with [ 3 H]Ro15–1788 (87 Ci/mmol; NEN Life Sciences), [ 3 H]Ro15–4513 (21.7 Ci/mmol; NEN Life Sciences) in the presence or absence of 10 μ m diazepam and [ 3 H]muscimol (19.1 Ci/mmol; NEN Life Sciences) were performed on membrane preparations as previously described ( Quirk et al., 1994 ; Sur et al., 1998 , 1999a ).

Techniques:

Journal: The Journal of Neuroscience

Article Title: Loss of the Major GABA A Receptor Subtype in the Brain Is Not Lethal in Mice

doi: 10.1523/JNEUROSCI.21-10-03409.2001

Figure Lengend Snippet: Generation-dependent upregulation of α2 and α3 subunits in cerebellum and forebrain of α1−/− mice.a, The loss of [3H]Ro15–1788 binding sites increased in cerebellum (left panel) and forebrain (right panel) of α1−/− mice from generation F2 to F5 to finally reach the proportion of [3H]Ro15–1788 sites immunoprecipitated by a selective α1 antibody from wild-type membrane (filled bars). ***p < 0.001; Student's t test. b, Evidence for an upregulation of α3 subunit in the cerebellum of α1−/− mice. Western blot showing an increase in α3 subunit expression in knock-out mice compared with wild-type (left panel). Quantitative immunoprecipitation with a α3 antibody demonstrated that α3 subunit-containing GABAAreceptors account for 14 ± 3% (mean ± SEM;n = 3) and 92 ± 8% (mean ± SEM;n = 4) of total [3H]Ro15–1788 binding sites in the cerebellum of wild-type and α1−/− mice, respectively (right panel). c, Quantitative immunoprecipitation demonstrated a 42% increase in the expression of α2 and α3 subunit-sensitive [3H]Ro15–1788 binding sites in the forebrain of α1−/− compared with wild-type mice.

Article Snippet: Radioligand binding assays with [ 3 H]Ro15–1788 (87 Ci/mmol; NEN Life Sciences), [ 3 H]Ro15–4513 (21.7 Ci/mmol; NEN Life Sciences) in the presence or absence of 10 μ m diazepam and [ 3 H]muscimol (19.1 Ci/mmol; NEN Life Sciences) were performed on membrane preparations as previously described ( Quirk et al., 1994 ; Sur et al., 1998 , 1999a ).

Techniques: Binding Assay, Immunoprecipitation, Western Blot, Expressing, Knock-Out

Journal:

Article Title: Mice devoid of ?-aminobutyrate type A receptor ?3 subunit have epilepsy, cleft palate, and hypersensitive behavior

doi:

Figure Lengend Snippet: Binding of [ 3 H]muscimol and [ 3 H]Ro15-4513 to whole brain homogenates prepared from postnatal day 1 (pups) or 8- to 12-week-old (adult) mice

Article Snippet: The benzodiazepine sites were labeled by [ 3 H]Ro15-4513 (10 nM; DuPont/NEN) ( 17 ) with and without zolpidem (Synthelabo Recherche, Bagneux, France) during a 60-min incubation at 0–4°C.

Techniques: Binding Assay

Journal:

Article Title: Mice devoid of ?-aminobutyrate type A receptor ?3 subunit have epilepsy, cleft palate, and hypersensitive behavior

doi:

Figure Lengend Snippet: Autoradiographic binding of a GABA site ligand ([3H]muscimol) and a benzodiazepine site ligand ([3H]Ro15-4513) to representative horizontal brain sections from newborn wild-type (+/+), heterozygous (+/−), and homozygous (−/−) mice. (A) Total binding of 5 nM [3H]Ro15-4513. (B) Binding of [3H]Ro15-4513 in the presence of 30 μM zolpidem. (C) Total binding of 20 nM [3H]muscimol. All images represent specific binding since [3H]Ro15-4513 and [3H]muscimol failed to label any sites in the presence of 10 μM flumazenil and 100 μM GABA, respectively.

Article Snippet: The benzodiazepine sites were labeled by [ 3 H]Ro15-4513 (10 nM; DuPont/NEN) ( 17 ) with and without zolpidem (Synthelabo Recherche, Bagneux, France) during a 60-min incubation at 0–4°C.

Techniques: Binding Assay

Journal:

Article Title: Mice devoid of ?-aminobutyrate type A receptor ?3 subunit have epilepsy, cleft palate, and hypersensitive behavior

doi:

Figure Lengend Snippet: Binding of [ 3 H]muscimol and [ 3 H]Ro15-4513 to postnatal day 1 mouse brain sections as revealed by autoradiography

Article Snippet: The benzodiazepine sites were labeled by [ 3 H]Ro15-4513 (10 nM; DuPont/NEN) ( 17 ) with and without zolpidem (Synthelabo Recherche, Bagneux, France) during a 60-min incubation at 0–4°C.

Techniques: Binding Assay