In all experiments, recording aCSF contained atropine (400 nM; Sigma-Aldrich) to block the actions of muscarinic ACh receptors when ACh was applied, or in case of nicotine application, to prevent muscarinic receptor activation by nicotine-induced endogenous ACh release68 (link). This ensured only the actions of nAChRs were measured and provided standardized background experimental conditions for both nicotine and ACh experiments. Other bath-applied drugs were similarly dissolved in aCSF at the desired concentration (nicotine (300 nM in tLTP experiments, 10 μM in two-photon imaging experiments; Sigma-Aldrich), ACh (1 mM; Sigma-Aldrich), Galanthamine (0.1 or 1 μM; Tocris Bioscience), DHβE (10 μM; Tocris Bioscience)). All experiments were performed in the absence of synaptic blockers, except the experiments with light-evoked endogenous ACh release on L2/3 interneurons (Fig. 2a , top traces), the DHβE pharmacology on L6 pyramidal neurons (Fig. 2b ), the experiments investigating dendritic expression of nAChRs (Fig. 4 ) and the DHβE pharmacology experiments on human L6 pyramidal neurons (Fig. 6d ), where GABAzine (10 μM; Tocris Bioscience) and DNQX (10 μM; Tocris Bioscience) were included in the aCSF. In the BAPTA-tLTP experiments (Fig. 5a,b ), BAPTA (1 mM; Sigma-Aldrich) was added to the intracellular solution.
Bapta
Manufactured by Merck Group
Sourced in United States, Germany, Italy, China
BAPTA is a laboratory reagent used as a calcium chelator. It has a high affinity for calcium ions and is commonly used in various research and experimental applications to regulate or manipulate calcium levels in biological systems.
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Lab products found in correlation
Ethylene glycol tetraacetic acid (egta), by Merck Group (8 mentions)
Picrotoxin, by Merck Group (6 mentions)
2 apb, by Merck Group (5 mentions)
Bapta am, by Merck Group (4 mentions)
D ap5, by Bio-Techne (4 mentions)
Latrunculin a, by Merck Group (3 mentions)
Na3gtp, by Merck Group (3 mentions)
Na2atp, by Merck Group (3 mentions)
Bicuculline, by Merck Group (3 mentions)
U73122, by Merck Group (3 mentions)
Thapsigargin, by Bio-Techne (2 mentions)
U73122, by Bio-Techne (2 mentions)
Kn 62, by Merck Group (2 mentions)
Sucrose, by Merck Group (2 mentions)
Carvacrol, by Merck Group (2 mentions)
Bapta am pf 127, by Merck Group (2 mentions)
Anti ha antibody, by Roche (2 mentions)
Nifedipine, by Merck Group (2 mentions)
L name, by Merck Group (2 mentions)
ω conotoxin mviic, by Bio-Techne (2 mentions)
75 protocols using bapta
1
Standardized nAChR Activation Experiments
2
Pharmacological Modulation of Neuronal Signaling
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Tetrodotoxin citrate (TTX), (−)-Bicuculline methiodide (BIC), 4-Aminopyridine (4-AP), Strychnine hydrochloride, Glycine (all Abcam), (2 R)-amino-5-phosphonovaleric acid; (2 R)-amino-5-phosphonopentanoate (D-APV) (R&D Systems), Phorbol-12-myristat-13-acetat (PMA), 1,2-bis(o-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid) (BAPTA) (Sigma).
3
Electrophysiological Recordings with Diverse Solutions
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Normal aCSF was used in most cases for in vitro electrophysiological recordings. Ca2+-free solution was made by removing Ca2+ chloride from the recording solution and replacing it with an equimolar concentration of magnesium chloride. Low-Na+ solution was made by substituting equimolar concentrations of Na+ chloride by choline chloride. All solutions were oxygenated with 95% O2/5% CO2. All salt compounds, TEA (#T2265); TPPO (#T84603), L.A. (#L1376), BAPTA (#A9801); Caffeine (#C0750), NMA (#M2137), and 5-HT (#S2805) were obtained from Sigma-Aldrich. TTX (#1078), Chelerythrin (#1330), U73122 (#1268), Xestospongin-C (#1280), 9-Phenanthrol (#4999), Dantrolene (#0507), and Thapsigargin (#1138) were obtained from Tocris Bioscience. Dantrolene, Thapsigargin, Xestospongin-C, U73122, and 9-Phenantrol were dissolved in dimethylsulfoxide (DMSO) and added to the aCSF (final concentration of DMSO: 0.05–0.1%). L.A. was dissolved in ethanol and added to aCSF (final concentration of ethanol: 0.05–0.1%). Control experiments showed no effects of the vehicle. The other drugs were dissolved in water and added to the aCSF.
4
Inhibition Assay for Cell Motility
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Latrunculin A (Sigma–Aldrich), 5‐(N‐Ethyl‐N‐isopropyl) amiloride (EIPA, Sigma–Aldrich), Ouabain (Sigma–Aldrich), Bumetanide (Ro 10–6338, Santa Cruz Biotechnology), Rab7 inhibitior CID 1067700 (Sigma–Aldrich), MyoVin‐1 (Sigma–Aldrich), BAPTA (Sigma–Aldrich), BAPTA‐AM (Sigma–Aldrich), FTY720, 2‐APB and CAI (Tocris Biosciences), Calpain inhibitor I (Millipore Sigma) in dimethylsulfoxide and Gadolinium (III) Chloride (Sigma–Aldrich) in water were used. For all inhibition experiments, cells were incubated in cell culture media with drugs for a duration of 1 h (except Ouabain ≈2 h) and then, high viscosity media was added. Cells were then imaged over a span of 5 h for speed measurements or were incubated over a span of ≈4 h for immunostaining.
5
Pharmacological Dissection of Synaptic Plasticity Mechanisms
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The following reagents (final concentration) were included in this study. DHβE (1 μM), MLA (10 nM), APV(50 μM), IEM-1460(50 μM), Ro318220(10 μM), Bicuculine (10 μM), TTX(1 μM) were from TOCRIS. (−)-nicotine (N3876, covered with dark paper when in use), U0126 (10 μM), FK506 (10 μM), KN62 (15 μM), H89 (10 μM), BAPTA (10 mM) and spermine (100 μM) were from Sigma. Lipid solvent were made in stock solutions in DMSO (1:1000–2000), while the same dilution of DMSO were used in control solutions. Peptides that were designed to block the phosphorylation of GluA1 at the PKA (S845), CAMKII (S831) and PKC (S816/818) sites were synthesized and purified by Pepmic Co., Ltd (Suzhou, China). The sequences of PKA S845 was TLPRNSGAG (-SGAG), scrambled control was NRPGGTLSA (-TLSA). That of CaMKII S831 was QSINEAIRTSTLPRN (-LPRN), scrambled control was NLIITEQRPSSNART (-NART). And that of PKCS816/818 was EFCYKSRSESKRMKGFC (-KGFC), scrambled control was RGMKESSKKCCRSFFYE (-FFYE). They were all diluted in internal solution with a final concentration of 200 μM.
6
Barium Current Recording Protocol
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The bath solution used to record Ba2+ currents contained (in mM) 10 BaCl2, 150 NaCl, 1 MgCl2, 10 HEPES, and 8 glucose (adjusted to pH 7.4 with NaOH). The pipette solution contained (in mM) 175 CsCl2, 5 MgCl2, 5 HEPES, 0.1 1,2-bis(2-aminophenocy)ethane N,N,N′,N′-tetraacetic acid (BAPTA), 3 Na2 ATP, and 0.1 Na3GTP (adjusted to pH 7.4 with CsOH). The external solution for confocal imaging contained (in mM) 160 NaCl, 2.5 KCl, 2 CaCl2 H2O, 1 MgCl2, 10 HEPES, and 8 glucose (adjusted to pH 7.4 with NaOH). The bath solutions were stored in a refrigerator at 4°C. The pipette solution was stored in a freezer at −20°C. BAPTA, Na2ATP, Na3GTP, CsOH, and BaCl2 reagents were obtained from Sigma-Aldrich (USA), HEPES was from Calbiochem (USA), and other chemicals were obtained from Merck (Germany).
7
Calcium Imaging of Cells with BAPTA
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Cells were grown in 35 mm glass-bottom dishes and were pre-treated with 50 μM of BAPTA (Sigma, St. Louis, MO) or PBS control for 30 min. Cells were then washed with OPTI-MEM followed by incubating with 5 μM Fluo-4/AM (Invitrogen, Carlsbad, CA) in the dark for 30 min in OPTI-MEM. After washing, the cellular fluorescence was imaged by a time lapse assay using confocal microscopy (LSM 510 meta; Carl Zeiss, Thornwood, NY).
8
Calcium Signaling in Drosophila Oocyte Activation
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BAPTA (Sigma-Aldrich) was used at a final concentration of 10 µM; BAPTA-AM + PF-127 (Sigma-Aldrich) was used at a final concentration of 30 µM; carvacrol (Sigma-Aldrich) used at 300–700 µm; copper sulfate (Sigma-Aldrich) was used at a final concentration of 2 mm. Standard preparation protocols were used as according to Sigma-Aldrich.
AB, 260 mOsm, containing 3.3 mm NaH2PO4, 16.6 mm KH2PO4, 10 mm NaCl, 50 mm KCl, 5% polyethylene glycol 8000, 2 mm CaCl2, brought to pH 6.4 with a 1 : 5 ratio of NaOH : KOH [15 (link)]; Gibco Schneider's Drosophila Medium, approximately 360 mOsm, a standard Drosophila culture medium that does not initiate egg activation (Thermo Fisher); Series95 halocarbon oil (KMZ Chemicals); EZ-Squeeze tube 125 µm (Cooper Surgical). For osmolarity experiments, sucrose (Sigma-Aldrich) was directly dissolved into distilled water and the osmolarity was measured using an osmometer (Löser).
AB, 260 mOsm, containing 3.3 mm NaH2PO4, 16.6 mm KH2PO4, 10 mm NaCl, 50 mm KCl, 5% polyethylene glycol 8000, 2 mm CaCl2, brought to pH 6.4 with a 1 : 5 ratio of NaOH : KOH [15 (link)]; Gibco Schneider's Drosophila Medium, approximately 360 mOsm, a standard Drosophila culture medium that does not initiate egg activation (Thermo Fisher); Series95 halocarbon oil (KMZ Chemicals); EZ-Squeeze tube 125 µm (Cooper Surgical). For osmolarity experiments, sucrose (Sigma-Aldrich) was directly dissolved into distilled water and the osmolarity was measured using an osmometer (Löser).
9
Pharmacological Modulation of Synaptic Responses
10
Ethanol Effects on Cytotrophoblast Cells
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