Stock solutions of 5-CT, BaCl2, tertiapin-Q, SCH23390 [(R)-(+)-7-Chloro-8-hydroxy-3-methyl-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine hydrochloride] were prepared in water and those of SCH28080 (2-Methyl-8-(phenylmethoxy)imidazo[1,2-a]pyridine-3-acetonitrile) and U73343 (1-[6-[[(17β)-3-Methoxyestra-1,3,5(10)-trien-17-yl]amino]hexyl]-2,5-pyrrolidinedione) in DMSO. All stock solutions, which were at least a thousand times the highest experimental concentration, were aliquoted and stored at -20°C until use. The highest experimental concentration of DMSO was 0.05%. 5-CT, SCH23390 and U73343 were purchased from Tocris (Tocris Bioscience, Bristol, UK); SCH28080 from HelloBio (Bristol, UK); CGP-55845; D -AP5, SR-95531, NBQX from Abcam (Cambridge, U.K.); tertiapin-Q from Abcam and Tocris; Isoflurane from Baxter S.p.A. (Rome, Italy); HEPES, ATP and DMSO from Fluka (St. Gallen, Switzerland). All other substances were obtained from Sigma-Aldrich (Milano, Italy)
Sr 95531
Manufactured by Abcam
Sourced in United Kingdom
SR-95531 is a GABA(A) receptor antagonist that selectively binds to and blocks GABA(A) receptors. It is used in research applications to study the function and pharmacology of GABA(A) receptors.
Automatically generated - may contain errors
Lab products found in correlation
D ap5, by Abcam (2 mentions)
Cgp55845, by Abcam (2 mentions)
U73343, by Bio-Techne (1 mentions)
Sch23390, by Bio-Techne (1 mentions)
Tertiapin q, by Abcam (1 mentions)
Gibco fbs, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Merck Group (1 mentions)
Nicl2, by Merck Group (1 mentions)
Bacl2, by Merck Group (1 mentions)
Xe991, by Abcam (1 mentions)
Zd7288, by Abcam (1 mentions)
Mefloquine hydrochloride, by Merck Group (1 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (1 mentions)
Qx 314 chloride, by Abcam (1 mentions)
Dmso dimethyl sulfoxide, by Merck Group (1 mentions)
2 3 dioxo 6 nitro 1 2 3 4 tetrahydrobenzo f quinoxaline 7 sulfonamide nbqx, by Bio-Techne (1 mentions)
Minocycline, by Abcam (1 mentions)
Cgp55845, by Bio-Techne (1 mentions)
Dcg 4, by Bio-Techne (1 mentions)
Naspm, by Bio-Techne (1 mentions)
6 protocols using sr 95531
1
Preparation of Pharmacological Reagents
2
Cell Culture and GABA Treatments
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The following chemicals and reagents were obtained from the indicated sources: fetal bovine serum (Gibco FBS; Invitrogen, Carlsbad, CA, USA); Dulbecco's modified Eagle's medium (DMEM), penicillin–streptomycin, and GABA (Sigma‐Aldrich Company, St. Louis, MO, USA); muscimol and GABAA antagonist (SR95531; Abcam Inc., Cambridge, MA, USA).
3
Pharmacological Isolation of Neuronal Signals
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For all experiments, external AFSF saline included 20 µM 6,7 dinitroquinoxaline-2,3-dione (DNQX), 25 µM D-22-amino-5 phosphonovaleric acid (D-APV), which were obtained from Alomone Labs, and 2 µM SR-95531 (gabazine; Abcam). In some experiments, 10 µM ZD7288, 10 µM XE991, 2 µM CGP-55845, or 0.5 µM TTX was included in the ACSF (Abcam). In addition, for some experiments, 2 mM NiCl2 or 50 µM BaCl2 (Sigma-Aldrich) was included in the ACSF. ZD7288 was only introduced through the bath transiently, for 3–4 min. This prevented a nonspecific depolarization that occurs with continuous bath application, yet provided a stable block for ∼30 min (Kim and Johnston, 2015 (link)).
4
Pharmacological Agents for Neuronal Studies
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D-AP5 (D-(-)−2-Amino-5-phosphopentanoic acid), SR 95531 (2–3-Carboxyprobyl)−3-amino-6(4-methoxyphenyl pyridazinium bromide), and QX-314 chloride were purchased from Abcam, Cambridge, UK. NBQX (2,3-Dioxo-6nitro-1,2,3,4-tetrahydrobenzo[f]quinoxaline-7-sulfonamide) was purchased from Tocris Bioscience, Bristol, UK. Alexa Fluor 488 and 594 were purchased from Life Technologies, Carlsbad, California, USA. Mefloquine hydrochloride and DMSO (Dimethyl sulfoxide) were bought from Sigma-Aldrich, St Louis, Missouri, USA.
5
Neurotransmitter modulation in cellular experiments
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DCG-IV (10 μM), 3,5-Dihydroxyphenylglycine (DHPG) (50 μM), and L-AP4 (200 μM) were puffed onto cells. LY341495 was both bath-applied (0.25–1 μM) and injected intraperitoneally (1–4 mg/kg) into mice for in vivo experiments. All other drugs were bath-applied: NBQX (10 μM), D-AP5 (50 μM), CGP55845 (1 μM), CGS15943 (0.5 μM), TBOA (100 μM), NASPM (50 μM), SR95531 (2–5 μM), minocycline (50 nM), Zn2+ (a free concentration of 300 nM was obtained by adding 60 μM Zn2+ and 10 mM tricine; Vergnano et al., 2014 (link)), Ro25-6981 (1 μM), and CIQ (20 μM). NBQX, APV, SR95531, and minocycline were from Abcam (UK). DCG-IV, DHPG, CGP55845, CGS15943, TBOA, NASPM, L-AP4, and LY341495 were from Tocris Bioscience (UK). All other drugs were from Sigma-Aldrich (France). Ro25-6981 was a kind gift from P. Paoletti (IBENS, Paris).
6
Optogenetic Dissection of OT-VP/LH Circuits
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Male A 2A R-Cre mice were anesthetized with pentobarbital (50 mg/kg, i.p.) four weeks after being infused with AAV-hSyn-DIO-ChR2-mCherry. Brain slices containing the OT, ventral pallidum (VP) and lateral hypothalamus (LH) were obtained as descried above.
We first examined the electrophysiological property of the recording neurons in the OT in current-clamp by injection of step currents. The photostimulation (5-ms 473-nm light pulses) to activate ChR2 was delivered at the OT via LED through the 40X objective lens.
Evoked postsynaptic currents from VP or LH neurons were triggered by photostimulation
(5-ms 473-nm light pulses) delivered at 10 Hz at the terminals of OT A 2A R neurons.
ACSF with the GABA A receptor antagonist SR-95531 (5 µM, abcam Biochemicals, UK)
or the AMPA/NMDA receptor antagonists NBQX (5 µM, Tocris Bioscience, UK) and D-APV (25 µM, Tocris Bioscience, UK) were applied to the VP or LH slice to clarify the receptor type of the postsynaptic currents.
We first examined the electrophysiological property of the recording neurons in the OT in current-clamp by injection of step currents. The photostimulation (5-ms 473-nm light pulses) to activate ChR2 was delivered at the OT via LED through the 40X objective lens.
Evoked postsynaptic currents from VP or LH neurons were triggered by photostimulation
(5-ms 473-nm light pulses) delivered at 10 Hz at the terminals of OT A 2A R neurons.
ACSF with the GABA A receptor antagonist SR-95531 (5 µM, abcam Biochemicals, UK)
or the AMPA/NMDA receptor antagonists NBQX (5 µM, Tocris Bioscience, UK) and D-APV (25 µM, Tocris Bioscience, UK) were applied to the VP or LH slice to clarify the receptor type of the postsynaptic currents.
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