We used optical stimulation to activate iMSNs selectively. Channelrhodopsin-2 (ChR2) and its reporter gene enhanced yellow fluorescent protein (EYFP) were inserted in a double-floxed inverted open reading frame viral vector and stereotaxically injected into the striatum of 1 month-old WT and R6/2 mice crossed with A2A-Cre mice (1.0 mm anterior and 2.0 mm lateral to Bregma, at a depth of 3.1 mm from the dura). After Cre recombination, ChR2-EYFP was selectively expressed in A2A-Cre neurons. To examine evoked GABA responses in GPe neurons the following protocol was used: a slow ramp voltage command (3 s duration) from a holding voltage of −70 mV to +10 mV was first applied. Then the cell was held at the new voltage for 2 s, after which the optical stimulation (470 nm, 1 mW, 0.5 ms) was delivered. After 2 additional s at +10 mV the membrane potential was brought back to the original holding potential. This protocol was repeated 3 times (interval between sweeps was 30 s) and the responses to light stimulation were averaged. SNAP5114 (GAT-3 antagonist, 20 μM), NO711 (GAT-1 antagonist, 10μM), or CGP 54626 (GABAB receptor antagonist, 1μM) (all from Tocris, Minneapolis, MN) were bath applied (10-20 min) to examine alterations in decay time (90%-10%) of the evoked GABA response measured with Clampfit v10.7 (Molecular Devices, Sunnyvale, CA; RRID:SCR_011323).
No 711
Manufactured by Bio-Techne
Sourced in United Kingdom
The NO-711 is a lab equipment product from Bio-Techne. It is designed for the detection and measurement of nitric oxide (NO) in various biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Snap 5114, by Bio-Techne (2 mentions)
Clampfit v10, by Molecular Devices (1 mentions)
Cgp54626, by Bio-Techne (1 mentions)
Axopatch 200b amplifier, by Molecular Devices (1 mentions)
Digidata 1322a, by Molecular Devices (1 mentions)
Bicuculline methiodide, by Merck Group (1 mentions)
Trkb fc, by R&D Systems (1 mentions)
1 3 butanediol, by Merck Group (1 mentions)
Clozapine n oxide, by Bio-Techne (1 mentions)
Bicuculline methiodide bmi, by Bio-Techne (1 mentions)
6 protocols using no 711
1
Optogenetic Manipulation of Striatal iMSN
2
Patch-clamp Recording of Hippocampal Neurons
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Patch-clamp recordings were obtained in acutely prepared coronal hippocampal slices from male rats, as described previously [6 (link)18 (link)]. Briefly, slices were perfused with artificial cerebrospinal fluid (aCSF; in mM: NaCl 126, KCl 2.5, MgSO4 1, NaHCO3 26, NaH2PO4 1.25, glucose 20, ascorbic acid 0.4, CaCl2 1, pyruvic acid 2; pH 7.3~7.4; saturated with 95%O2–5%CO2) at a ~3 ml/min flow. Recordings were obtained at 32℃ using an Axopatch 200B amplifier (Axon Instruments, Foster City, CA). The series resistance was motored throughout the experiments. Neurons localized in the outer half of the granule cell layer were selected to minimize the effects of neurogenesis [19 (link)]. Patch pipettes were filled with a high Cl− containing solution (in mM): KCl 140, HEPES 10, Mg2+ATP 5, MgCl2 0.9, and EGTA 10. Current output was filtered at 2 kHz and digitized at 10 kHz (Digidata 1322A, pClamp 9 software, Axon Instruments). Itonic was defined as the difference between the holding current (Iholding) before and after application of the GABAA receptor blocker bicuculline (20 µM). Drugs were added to the perfusing aCSF solution at known concentrations. All drugs except NO-711 (Tocris, UK) were purchased from Sigma-Aldrich (St. Louis, MO, USA).
3
Targeted Drug Delivery for Electrophysiology
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Frozen DMSO stocks of NO711 (Tocris) were diluted 1000-fold in extracellular solution on the day of recording. Drug was gravity fed through a 0.5 mm ID Teflon tube targeted at the cell being recorded, which was switched between control and drug solution using a two-way valve upstream of the bath (exchange time ~10 s). Background bath perfusion of control solution helped to minimize the drug effect on surrounding cells.
4
Pharmacological Modulation of GABA Transporters
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Bicuculline methiodide was purchased from Sigma-Aldrich (St. Louis, MO). The GAT1 blocker NO-711 [(1,2,5,6-tetrahydro-1-[2-[[(diphenylmethylene)amino]oxy]ethyl]-3-pyridinecarboxylic acid hydrochloride)] and GAT3 blocker SNAP-5114 [1-[2-[tris(4-methoxyphenyl)methoxy]ethyl]-(S)-3-piperidinecarboxylic acid] were purchased from Tocris Bioscience (Minneapolis, MN).
5
Intracerebral Infusion of Neuroprotective Agents
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Immediately after photothrombotic stroke was induced, 26-gauge, 3.5 mm guide cannulae (RWD Life Sience) were implanted into the core of the infarction (anteroposterior: 0 mm; mediolateral: -1.5 mm; dorsoventral: -1.0 mm; relative to the bregma). The cannulae were fixed to the skull using adhesive luting cement and acrylic dental cement. Following surgery, stainless steel obturators were inserted into the guide cannulae to avoid obstruction until infusions were made. For drug infusions, obturators were removed and 33-gauge, 4.0 mm injectors (RWD Life Science) were placed into the guide cannulae. Injector tips extended 0.5 mm beyond the guide cannulae. During drug injection, 10 μM NO-711 (Tocris Bioscience), or 1 mg/mL TrkB-Fc (R&D system) were slowly infused at a flow rate of 0.2 μL/min to a total volume of 2 μL. Following infusion, the injectors were left in place for an additional 5 min to allow for diffusion of the drug. Following microinjection, the stainless steel obturators were subsequently reinserted into the guide cannulae.
6
Electrophysiological Drug Compounds Protocol
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Drugs NO-711 (Tocris Bioscience; cat #1779), clozapine-N-oxide (CNO, Tocris Bioscience; cat #4936), bicuculline methiodide (BMI, Tocris Bioscience; cat #0109), 1,3-butanediol (Sigma-Aldrich; cat #309443), b-hydroxybutrate (b-HB, Sigma-Aldrich; cat #298360). All drugs used in electrophysiological experiments were supplied by Tocris Bioscience or Sigma-Aldrich.
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