To examine the role of local inhibition in controlling spiking reliability in the network, we used data from extracellular recordings in NCM collected as a part of previous experiments (Thompson et al., 2013 (link)). Briefly, commercial multibarreled glass pipettes containing a carbon fiber electrode (5 μm diameter; 400–1,200 kΩ impedance) and six attached barrels (~3 μm diameter) were used for drug microiontophoresis (Kation Scientific, Minneapolis, MN). Gabazine (SR95531, 3 mM, pH 3.2; Sigma Aldrich, St. Louis, MO), or a Gabazine/saclofen combination, was used to inhibit GABA-mediated inhibition locally in NCM around the recording site. The combined application of Gabazine and saclofen (n = 12 sites) did not elicit responses different from those during application of Gabazine alone (Thompson et al., 2013 (link)). Following Thompson et al (2013) (link) we considered drug delivery at a particular site successful if the average song-driven firing rate during iontophoresis was significantly different (either higher or lower) from the firing rate prior to iontophoresis.
Gabazine
Manufactured by Merck Group
Sourced in United States, China
Gabazine is a laboratory product manufactured by Merck Group. It is a GABA receptor antagonist used for research purposes.
Automatically generated - may contain errors
Lab products found in correlation
Strychnine, by Merck Group (11 mentions)
Picrotoxin, by Merck Group (5 mentions)
Muscimol, by Merck Group (4 mentions)
Bumetanide, by Merck Group (4 mentions)
Sr95531, by Merck Group (4 mentions)
6 cyano 7 nitroquinoxaline 2 3 dione cnqx, by Merck Group (3 mentions)
Tetrodotoxin ttx, by Merck Group (3 mentions)
Cgp55845, by Merck Group (3 mentions)
6 ohda hydrochloride, by Merck Group (2 mentions)
Cgp54626, by Bio-Techne (2 mentions)
Bicuculline, by Bio-Techne (2 mentions)
Baclofen muscimol, by Merck Group (2 mentions)
Cnqx ap5, by Merck Group (2 mentions)
Isoguvacine, by Merck Group (2 mentions)
4 aminopyridine 4 ap, by Merck Group (2 mentions)
Glycine, by Merck Group (2 mentions)
Baclofen, by Merck Group (2 mentions)
Saclofen, by Merck Group (2 mentions)
Bicuculline, by Merck Group (2 mentions)
Mecamylamine hydrochloride, by Merck Group (2 mentions)
57 protocols using gabazine
1
Inhibiting GABA-mediated Spiking Reliability
2
Blocking GABAergic and Glutamatergic Signaling
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Bicuculline, a GABAA receptor blocker, was applied at a final concentration of 50–100 μM. Gabazine (SR-95531, Sigma), a reversible GABAA receptor blocker, was applied at a final concentration of 300 nM. CNQX, a glutamatergic AMPA receptor blocker, was applied at a final concentration of 25 μM. Bicuculline, Gabazine, and CNQX were added to the external Ringer solution and the recording started after approximately 30 min. In some experiments the recording electrode was filled with QX- 314, an intracellular Na+ channel blocker, (100 mM dissolved in 2 M KAc) or QX-314 and CsCl, an intracellular K+ channel blocker (1 M CsCl, 100 mM QX-314 and 1 M KAc). Positive current pulses of 0.5–1.2 nA in amplitude, 50–100 ms in duration repeated at 1 Hz were used to deliver the drugs to the recorded cell.
3
Intravitreal Injection of Neurotoxins in Zebrafish
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Different neurotoxins were injected into the vitreous using a protocol adapted from previous studies (Rajaram et al., 2014a (link), Rajaram et al., 2014b (link), Thummel et al., 2008 (link)). The drugs included gabazine (Sigma S106), NBQX (Abcam ab210046), muscimol (Sigma M1523), and AMPA (Sigma A9111). In brief, zebrafish were anesthetized in 0.016% tricaine, an incision was made in the sclera with a sapphire knife, and a blunt-end 30-gauge needle inserted. Drugs (0.5 μL) were injected into one eye of adult zebrafish. Fish were immediately placed into a recovery tank; times indicated are hours of recovery.
4
Investigating GABAergic Inhibition in Neural Responses
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To assess the role of GABAergic inhibition in neural responses, the
GABAA receptor antagonist gabazine (SR95531, Sigma-Aldrich) was
locally injected during neural recordings. For drug injection experiments, a
cannula was implanted along with the tetrodes. gabazine was used at
1 μM because that concentration did not induce seizure-like neural
activity in our preliminary experiments. The single-unit activity was isolated,
and auditory responses to the song stimuli were recorded. Then, a bird was
placed in the stereotaxic apparatus, and gabazine (200 nl of a
1 μM solution in saline) was locally injected through the guide
cannula over a period of 5 min (KDS 310 Nano Pump, KD Scientific). The bird was
returned to the recording chamber, and auditory responses of the same single
unit were again recorded at 20 min (gabazine condition) and at 1 h after
injection (recovery condition). In control experiments, the same amount of
saline was injected, and electrophysiological recordings were performed before
and after injection.
GABAA receptor antagonist gabazine (SR95531, Sigma-Aldrich) was
locally injected during neural recordings. For drug injection experiments, a
cannula was implanted along with the tetrodes. gabazine was used at
1 μM because that concentration did not induce seizure-like neural
activity in our preliminary experiments. The single-unit activity was isolated,
and auditory responses to the song stimuli were recorded. Then, a bird was
placed in the stereotaxic apparatus, and gabazine (200 nl of a
1 μM solution in saline) was locally injected through the guide
cannula over a period of 5 min (KDS 310 Nano Pump, KD Scientific). The bird was
returned to the recording chamber, and auditory responses of the same single
unit were again recorded at 20 min (gabazine condition) and at 1 h after
injection (recovery condition). In control experiments, the same amount of
saline was injected, and electrophysiological recordings were performed before
and after injection.
5
6-OHDA Induced Neurodegeneration Protocol
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Isoflurane was purchased from RWD Life Science (Shenzhen, China). Pentobarbital and lidocaine were purchased from Chaohui Pharmaceutical. 6‐OHDA, AP‐5, DNQX, strychnine, muscimol, and gabazine were purchased from Sigma‐Aldrich. CY3 Donkey anti‐rabbit IgG, Mouse and Rabbit Specific HRP/DAB (ABC), Detection IHC kit, and rabbit antityrosine hydroxylase are products of Abcam Corp. 6‐OHDA hydrochloride (Sigma) was dissolved in 0.9% saline containing 0.2% ascorbic acid to avoid decomposition.
6
Colon Motor Activity Analysis in Mice
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Adult male NL3R451C and WT mice (25–35 g; 8–12 weeks of age) were euthanized by cervical dislocation. The entire colon (~5 cm in length) was dissected, flushed, and cannulated in an organ bath superfused with physiological saline (at 37°C, 95%O2/5% CO2, flow: 5 mL min−1). After a 30 min equilibration, video recordings were acquired using a Logitech Quickcam pro 9000 camera (30 fps, 640 × 840 resolution) in order to measure contractile motor activity for a duration of 3 hr. Motor activity was recorded at baseline (1 hr), with bath‐applied antagonist (1 hr), and during washout (1 hr). Pseudocolored spatiotemporal maps of motor patterns were created and motor pattern frequencies were quantified using in‐house software [Neal et al., 2009 ; Swaminathan et al., 2016 ] (Fig. S1 ). Bicuculline (10 μM, Tocris), gabazine (10 μM, Sigma), and CGP 54626 (100 nM, Tocris) were diluted in saline.
7
Neurotransmitter Receptor Antagonist Application
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Drugs were prepared as concentrated stock solutions and diluted in ACSF for bath application immediately prior to use. Gabazine, a selective GABAA receptor antagonist; (±)-2-amino-5-phosphonopentanoic acid (AP5, a selective NMDA receptor antagonist), 6-cyano-7nitroquinoxaline 2, 3-dione (CNQX, an AMPA-KA receptors antagonist), DL-Serine 2-(2,3,4-trihydroxybenzyl)hydrazide hydrochloride (benserazide), an inhibitor of peripheral dopa decarboxylase and α-methyl-L-tyrosine (levodopa) were all purchased from Sigma Aldrich (France).
8
Comprehensive Pharmacology Reagents Protocol
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We purchased common salts and 1-(3-Chlorophenyl) biguanide hydrochloride (mCPBG); gabazine; 1,2-bis(2-aminophenoxy) ethane-N,N,N,N-tetraacetic acid (BAPTA); SCH50911; 6,7-dinitroquinoxaline-2,3-dione (DNQX); tetrodotoxin (TTX); strychnine; SB200646 hydrochloride (SB200646); ritanserin (RIT); ondansetron hydrochloride dihydrate (OND); and 5-HT hydrochloride (5-HT) from Sigma-Aldrich Chemical Company (St Louis, MO, USA). Citalopram hydrobromide, WAY100635, and 1-(3-Chlorophenyl) piperazine hydrochloride (mCPP) from Tocris Bioscience (Ellisville, MO, USA).
9
Preparation of Neurotransmitter Solutions
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Diacetyl (Sigma-Aldrich) was diluted in water to reach the 1:1000 ratio. Glycerol (BP229-1, Fisher Scientific) was diluted to final concentrations in a buffer containing 100-mM NaCl, 10-mM KCl, and 30-mM Tris [PH 7.5]. ACh (AC159170050, ACROS Organics) and gabazine (S106, Sigma-Aldrich) were first dissolved in water to make aliquots of 10-mM frozen stocks, which were diluted to final concentrations using the bath solution before use.
10
Electrophysiology Reagent Preparation
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