Cgp35348
Manufactured by Bio-Techne
Sourced in United Kingdom
CGP35348 is a chemical compound used in laboratory research. It serves as a GABAB receptor antagonist, which means it blocks the activity of GABAB receptors. The compound is typically used in in vitro and in vivo studies to investigate the role of GABAB receptors in various biological processes.
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Lab products found in correlation
Baclofen, by Bio-Techne (3 mentions)
Ro 25 6981, by LC Laboratories (2 mentions)
Ro25 6981, by Bio-Techne (2 mentions)
Biocytin, by Merck Group (1 mentions)
Gabazine, by Merck Group (1 mentions)
Carbenoxolone, by Merck Group (1 mentions)
Gabazine, by Bio-Techne (1 mentions)
D ap5, by Bio-Techne (1 mentions)
Baclofen, by Merck Group (1 mentions)
Bicuculline methiodide, by Bio-Techne (1 mentions)
Rapamycin, by Bio-Techne (1 mentions)
Rapamycin, by LC Laboratories (1 mentions)
Beveled quartz micropipette, by Sutter Instruments (1 mentions)
Sr 95531 hydrobromide, by Bio-Techne (1 mentions)
Lidocaine, by Merck Group (1 mentions)
Cnqx disodium salt, by Bio-Techne (1 mentions)
R 5000, by LC Laboratories (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Muscimol, by Bio-Techne (1 mentions)
Picrotoxin, by Bio-Techne (1 mentions)
9 protocols using cgp35348
1
Electrophysiology Reagent Preparation
2
Baclofen Dosage Regimen Post-TBI
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NS and Baclofen were administered by intraperitoneal injection (i.p.) for 3 consecutive weeks a day after TBI. Baclofen was purchased from Sigma Aldrich, UK (B5399) and dissolved in NS at a concentration of 0.05 mg/kg, 0.2 mg/kg, and 1 mg/kg. CGP35348 was obtained from Tocris, UK (1245) and dissolved in the same solvent. Baclofen concentrations were determined with reference to the clinical dosage. Mice were administered Baclofen a day after TBI induction for 3 consecutive weeks. After injections were completed, the drug was withdrawn from the mice, and they underwent additional observation periods for 3 weeks. They were then sacrificed at 6 weeks after CCI modeling.
3
Optophysiology with GABA Antagonists
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Drug manipulations to the slice during optophysiological experiments were administered via ACSF perfusion. Antagonists included the GABA-A blocker bicuculline methiodide (BCC, 3 μM, Tocris) by itself or, in a subset of neurons, BCC in combination with the GABA-B blocker CGP 35348 (CGP, 1 μM; Tocris). Antagonist was pre-applied for 5 minutes before and co-applied during the relevant optophysiological recording. To examine the recovery of the optophysiological effect upon partial washout, additional recordings were performed in a subset of neurons ∼5-10 mins after BCC application ended.
4
Baclofen's GABA-B Receptor Mechanism
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To verify that baclofen was acting through GABAB receptor mechanisms, an antagonist was administered IT prior to administration of baclofen. Specifically, these rats received repeated UBDs (60 mm Hg, 20 s 3 min ITI). After VMRs to UBD were stable (± 20%), responses to graded UBD (10–60 mm Hg, 20 s) were obtained. Rats then received a single IT injection of either CGP35348 (30 µg; Tocris Biosciences, Minneapolis, MN, USA; Cat. No. 1245) or an equal volume of normal saline (10 µl) followed by a 10 µl normal saline flush. Fifteen minutes after the injection repeat graded UBD stimulus–response measures were obtained. Rats then received an IT injection of baclofen (40 ng) in normal saline or an equal volume of normal saline (10 µl) followed by a 10 µl normal saline flush. Fifteen minutes after this injection, repeat graded UBD stimulus–response measures were obtained. These dosing combinations resulted in three separate measures in four separate groups.
5
Pharmacological Modulation of Synaptic Plasticity
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For behavioral and in vivo experiments, animals received intraperitoneal (i.p.) injections of Ro-25-6981 (10 mg/kg, Tocris, 1594) with or without CGP35348 (100 mg/kg; Tocris, 1245), or 0.9% physiological saline vehicle (0.01 ml/g, LabChem). Animals in the RNA immunoprecipitation (RIP)-Seq experiment received either Ro-25-6981 (10 mg/kg) with or without rapamycin (6 mg/kg; LC Laboratories, #R-5000) or 0.9% physiological saline (200 μl). For in vitro experiments, Ro-25-6981 (10 µM), rapamycin (200 nM), and baclofen (50 µM; Tocris, #0796) were used. Vehicle controls cells were treated with water (vehicle for Ro-25-6981 and baclofen) and/or 0.1% DMSO (vehicle for rapamycin).
6
Pharmacological Manipulation of Purkinje Neuron Physiology
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A micropipette inserted through the chronic cranial window access port was used for pharmacological manipulation. In this case, the micropipette: a beveled quartz micropipette (0.7 mm ID, Sutter Instruments) was placed ~50 μm below the PN soma for drug injections.
Lidocaine (2%) (Sigma) and CNQX disodium salt (100 μM) (Tocris) were used to block Na + channels and AMPA receptors (excitatory synaptic input) respectively. CGP 35348 (100 μM)
(Tocris) was used to block GABA B receptors. GABA B receptors are predominantly localized to the imaging region (PF-PN synapses in the spiny dendrites) (Kulik et al., 2002) , whereas GABA A receptors are expressed abundantly throughout the cerebellum, notably at basket cell terminals on the PN soma (Fritschy and Panzanelli, 2006) . Blocking GABA A receptors with SR 95531 hydrobromide (100 μM) (Tocris) led to over-excitability of the PN dendrites and irregular spontaneous spiking behavior. AIDA (100 μM) (Tocris) was used to block mGluR1 activity. All drugs were dissolved in saline and applied by pressure injection (<0.5 psi) for 10 min prior to imaging and reduced to <0.1 psi while dendritic voltage and calcium recordings were repeated in the awake mice. Dendritic recordings were occasionally repeated ~24h after drug application (guaranteeing drug washout) to confirm that the labelled PN was not physically damaged by the drug application.
Lidocaine (2%) (Sigma) and CNQX disodium salt (100 μM) (Tocris) were used to block Na + channels and AMPA receptors (excitatory synaptic input) respectively. CGP 35348 (100 μM)
(Tocris) was used to block GABA B receptors. GABA B receptors are predominantly localized to the imaging region (PF-PN synapses in the spiny dendrites) (Kulik et al., 2002) , whereas GABA A receptors are expressed abundantly throughout the cerebellum, notably at basket cell terminals on the PN soma (Fritschy and Panzanelli, 2006) . Blocking GABA A receptors with SR 95531 hydrobromide (100 μM) (Tocris) led to over-excitability of the PN dendrites and irregular spontaneous spiking behavior. AIDA (100 μM) (Tocris) was used to block mGluR1 activity. All drugs were dissolved in saline and applied by pressure injection (<0.5 psi) for 10 min prior to imaging and reduced to <0.1 psi while dendritic voltage and calcium recordings were repeated in the awake mice. Dendritic recordings were occasionally repeated ~24h after drug application (guaranteeing drug washout) to confirm that the labelled PN was not physically damaged by the drug application.
7
In Vivo and In Vitro Pharmacological Assays
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For behavioral and in vivo experiments, animals received intraperitoneal (i.p.) injections of Ro-25-6981 (10 mg/kg, Tocris 1594) with or without CGP35348 (100 mg/kg; Tocris, 1245), or 0.9% physiological saline vehicle (0.01 ml/g, LabChem). Animals in the RIP-Seq experiment received either Ro-25-6981 (10 mg/kg) with or without rapamycin (6 mg/kg; LC Laboratories, #R-5000), or 0.9% physiological saline (200 μl). For in vitro experiments, Ro-25-6981 (10 µM), rapamycin (200 nM), and baclofen (50 µM; Tocris, #0796) were used. Vehicle controls cells were treated with water (vehicle for Ro-25-6981 and baclofen) and/or 0.1% DMSO (vehicle for rapamycin).
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprint this version posted November 18, 2020. ; https://doi.org/10.1101/2020.08.25.266833 doi: bioRxiv preprint
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprint this version posted November 18, 2020. ; https://doi.org/10.1101/2020.08.25.266833 doi: bioRxiv preprint
8
Injection Procedure for Bee Neurotransmitter Receptors
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Muscimol, baclofen, picrotoxin and CGP35348 of >99.5% purity were obtained from Tocris Bioscience. The compounds were dissolved in 1 M dimethyl sulfoxide (DMSO; Sigma Aldrich) to make the working concentrations of 1.0, 0.10 and 0.01 mM used for injection. These concentrations of ligands were used as this range of agonists and antagonists were effective for other neurotransmitter receptors (Fussnecker et al., 2006; Mustard et al., 2010) . Individual bees were collected from the box into a plastic vial and cold anaesthetised prior to the injection procedure. Each bee was restrained in a metal harness, then a hole was made in the dorsal side of the thorax using a 19 gauge needle. A 10 µl Hamilton syringe was used to inject 1 µl of treatment solution. Control animals were injected with either 1 µl of double distilled water or 1 M DMSO; data from the water and DMSO injected animals was pooled for subsequent analyses, as there was no significant difference between the behaviors of bees in these groups.
9
Neurochemical Stock Solution Preparation
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Drugs were made up as stock solutions (1-10 mM) in distilled water and then diluted in aCSF.
Adenosine, 8-cyclopentyltheophylline (8CPT) were obtained from Sigma (Dorset, UK).
Tertiapin-Q and CGP35348 were obtained from (Tocris, Abingdon, UK).
Adenosine, 8-cyclopentyltheophylline (8CPT) were obtained from Sigma (Dorset, UK).
Tertiapin-Q and CGP35348 were obtained from (Tocris, Abingdon, UK).
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