L-Glutamate, D-glutamate, ivermectin and picrotoxinin (PTX) were obtained from Sigma-Aldrich (UK). Fipronil was a gift from Dr. Lance Hammerland (Merial Ltd). Kainic acid (referred to as kainate throughout this paper), N-methyl-D-aspartic acid (NMDA), quisqualic acid (referred to as quisqualate throughout this paper), L-aspartatic acid (referred to as aspartate throughout this paper), α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) were obtained from Tocris (UK), whereas ibotenic acid (referred to as ibotenate throughout this paper) was obtained from Wako Pure Chemical Industries (Osaka, Japan). Okaramine B was isolated from fermentation products of P. simplicissimum according to the original paper (Hayashi et al., 1989 ).
N methyl d aspartic acid
Manufactured by Bio-Techne
Sourced in United Kingdom
N-methyl-D-aspartic acid is a chemical compound commonly used in laboratory research. It is a selective agonist for the N-methyl-D-aspartate (NMDA) receptor, a type of glutamate receptor in the central nervous system. This product can be utilized in various experimental settings to study the role of NMDA receptors in cellular signaling and neurological processes.
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Lab products found in correlation
Kainic acid, by Bio-Techne (2 mentions)
L glutamate, by Merck Group (1 mentions)
Ivermectin, by Merck Group (1 mentions)
Picrotoxinin, by Merck Group (1 mentions)
D glutamate, by Merck Group (1 mentions)
Quisqualic acid, by Bio-Techne (1 mentions)
Ibotenic acid, by Fujifilm (1 mentions)
Glycine, by Bio-Techne (1 mentions)
Cnqx disodium salt, by Bio-Techne (1 mentions)
L glutamic acid, by Merck Group (1 mentions)
Kainic acid, by Fujifilm (1 mentions)
Y 27632, by Fujifilm (1 mentions)
Catalase, by Merck Group (1 mentions)
Anti glun2b, by Abcam (1 mentions)
Anti lamp1, by Abcam (1 mentions)
Anti actin, by Abcam (1 mentions)
Anti α synuclein, by BD (1 mentions)
Anti dapk1, by Merck Group (1 mentions)
Anti lamp2a, by Abcam (1 mentions)
Pepstatin a, by Merck Group (1 mentions)
5 protocols using n methyl d aspartic acid
1
Neurotransmitter Receptor Ligand Sources
2
Extracellular Solutions for Neuronal Recordings
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Extracellular solution (used for slicing and perfusion of slices): 125 mmol/L NaCl, 2.5 mmol/L KCl, 2 mmol/L CaCl2, 1 mmol/L MgCl2, 1.25 mmol/L NaH2PO4, 25 mmol/L NaHCO3, 25 mmol/L glucose, 10 μmol/L glycine (NMDAR coagonist, see Traynelis et al. 2010 ), bubbled with carbogen gas (95% O2, 5% CO2), pH 7.4 (c.f. Sakmann and Neher 1995 , p. 200; Spruston et al. 1995 ; Vargas‐Caballero and Robinson 2004 ).
Extracellular pipette solution: 145 mmol/L NaCl, 2.5 mmol/L KCl, 2 mmol/L CaCl2, 10 μmol/L glycine, 10 μmol/L CNQX (AMPA and kainate receptor blocker; CNQX disodium salt was obtained from Tocris), 10 mmol/L HEPES, pH was adjusted to 7.4 with about 4.2 mmol/L NaOH. (Mg‐free to prevent Mg‐block of NMDARs, see Cull‐Candy,2007 .)
L‐Glutamic acid was obtained from Sigma‐Aldrich. N‐Methyl‐D‐aspartic acid was obtained from Tocris.
Extracellular pipette solution: 145 mmol/L NaCl, 2.5 mmol/L KCl, 2 mmol/L CaCl2, 10 μmol/L glycine, 10 μmol/L CNQX (AMPA and kainate receptor blocker; CNQX disodium salt was obtained from Tocris), 10 mmol/L HEPES, pH was adjusted to 7.4 with about 4.2 mmol/L NaOH. (Mg‐free to prevent Mg‐block of NMDARs, see Cull‐Candy,
L‐Glutamic acid was obtained from Sigma‐Aldrich. N‐Methyl‐D‐aspartic acid was obtained from Tocris.
3
Excitotoxic Injury Model of Cochlear Tissue
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Damaged cochlear tissue were created as described elsewhere (Wang and Green, 2011 (link)). Following the original method, N-methyl-D -aspartic acid 0.5 mM (NMDA; Tocris Bioscience, Ellisville, MO, United States) 0.5 mM and kainic acid (Tocris Bioscience; NK treatment) were reacted in a culture solution for 2 h.
The following three groups were compared to investigate the effects of a ROCK inhibitor, Y-27632 (257-00511, Wako) in a model of excitotoxic injury of cochlear tissue: a control group; an NK group (NMDA 0.5 mM + kainic acid 0.5 mM for 2 h, followed by washing and culture in normal culture medium); and a ROCK inhibition group (washed after treatment with NK with addition of 10 μM Y-27632 in normal culture medium). Immunohistochemical evaluations were performed at 24 and 72 h after culture.
The following three groups were compared to investigate the effects of a ROCK inhibitor, Y-27632 (257-00511, Wako) in a model of excitotoxic injury of cochlear tissue: a control group; an NK group (NMDA 0.5 mM + kainic acid 0.5 mM for 2 h, followed by washing and culture in normal culture medium); and a ROCK inhibition group (washed after treatment with NK with addition of 10 μM Y-27632 in normal culture medium). Immunohistochemical evaluations were performed at 24 and 72 h after culture.
4
Immunoblotting and Immunoprecipitation Protocols
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Anti-GFP (Clonetech, 632381), anti-LAMP-1(Abcam, ab13523), anti-GAPDH (Abcam, ab9485), anti-actin(Abcam, ab8227), anti-α-synuclein (BD Transduction Laboratories, 610786), anti-HA (Roche applied science, 11867431001), monoclonal anti-FLAG M2 antibody (Sigma-Aldrich, F1804-200UG), anti-DAPK1 (Sigma, D1319-200UL), monoclonal anti-phospho-DAPK1 (pSer308, Sigma, D4941), anti-GluN2B (lab generated), anti-LAMP-2A (Abcam, ab18528), anti-Labmin B1(Abcam, ab16048), anti-HSP90 (BD Transduction Laboratories, 610418), anti-VDAC1 (Porin) (MitoSciences,MSA03). Antibodies were validated for their intended purpose (immunoblotting, immunocytochemistry, immunohistochemistry and co-immunoprecipitation) as outlined in the product sheet or in lab. Ammonium chloride (Sigma, A0171), 3-methyladenine (Sigma, M9281), MG132 (Sigma, C2211), Pepstatin A (Sigma), N-Methyl-D-aspartic acid (NMDA, Tocris Asc-052), H2O2 (Sigma, 7722-84-1) Catalase (Sigma, C1345), (2R)-amino-5-phosphon-opentanoic acid (APV, Ascent Scientific, Asc-003). GluN2B-CTM and TAT-GluN2B were synthesized by Brain Research Centre peptide synthesis facility at UBC. All other synthetic peptides used were synthesized by GL Biochem.
5
LFS-Induced Synaptic Plasticity Modulation
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For drug application, the aCSF solution was switched to the drug solution for at least 15 min before the induction of LFS. D-2-amino-5-phosphonovalerate (D-AP5), (S)-3,5-Dihydroxyphenylglycine ((S)-3,5-DHPG)), DL-threo-β-benzyloxyaspartate (DL-TBOA), (S)-(+)-α-Amino-4-carboxy-2-methylbenzeneacetic acid (LY367385), (+)-5-methyl-10,11-dihydroxy-5H-dibenzo(a,d)cyclohepten-5,10-imine[(+)-MK-801],N-[3-[[4-[(3-Aminopropyl)amino]butyl]amino]propyl]-1-naphthaleneacetamide (NASPM) trihydrochloride, N-Methyl-D-aspartic acid (NMDA), (αR,βS)-α-(4-Hydroxyphenyl)-β-methyl-4-(phenylmethyl)-1-piperidinepropanol (Ro 25-6981) maleate, and (1R*,2S*)-erythro-2-(4-Benzylpiperidino)-1-(4-hydroxyphenyl)-1-propanol (Ifenprodil) hemitartrate were obtained from Tocris Cookson (Bristol, UK). Aminoacetic acid, Aminoethanoic acid (Glycine) and [[[(1S)-1-(4-Bromophenyl)ethyl]amino](1,2,3,4-tetrahydro-2,3-dioxo-5-quinoxalinyl)methyl] phosphonic acid (PEAQX, NVP-AAM 077) tetrasodium hydrate were purchased from Sigma Aldrich (St. Louis, MO).
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