Kainate

Manufactured by Bio-Techne

Sourced in United Kingdom, United States

Kainate is a laboratory reagent used in neuroscience research. It is a potent agonist of the kainate subtype of ionotropic glutamate receptors, which play a role in excitatory neurotransmission in the central nervous system.

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Lab products found in correlation

Nbqx disodium salt, by Bio-Techne (3 mentions) Bicuculline, by Merck Group (2 mentions) Bicuculline, by Bio-Techne (2 mentions) Philanthotoxin, by Bio-Techne (2 mentions) Salts and general reagents, by Merck Group (2 mentions) Rp br camp, by Bio-Techne (2 mentions) Thapsigargin, by Bio-Techne (2 mentions) Forskolin, by Bio-Techne (2 mentions) Ryanodine, by Bio-Techne (2 mentions) Pentylenetetrazole, by Merck Group (2 mentions) Flurothyl, by Merck Group (2 mentions) Na3gtp, by Roche (2 mentions) R cpp, by Bio-Techne (2 mentions) Sch58261, by Bio-Techne (1 mentions) Picrotoxin, by Bio-Techne (1 mentions) Hamilton pipette, by Hamilton Company (1 mentions) Pertussis toxin, by Bio-Techne (1 mentions) Propidium iodide, by Thermo Fisher Scientific (1 mentions) Malachite green, by Merck Group (1 mentions) Ammonium molybdate, by Merck Group (1 mentions)

Spelling variants of this product

Kainate (KA; (3 citations) AMPA/Kainate receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX; (2 citations)

27 protocols using kainate

Kainate-Induced Seizure Modeling in Rodents

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In rats, kainate (Tocris) was injected intraperitoneally at a dose of 10 mg/kg, following our previous experience (Rebola et al., 2005a (link)). The selective A_2AR antagonist SCH58261 (Tocris) was used at an effective dose of 0.05 mg/kg (Lopes et al., 2004 (link)), administered intraperitoneally 30 min before kainate in most experiments, or 4 h after the extinction of convulsions in the last experimental protocol. A_2AR-KO and wild-type littermates or forebrain A_2AR-KO and floxed A_2AR-KO littermates (designated as wild-type), all with a C57BL6 background, were injected subcutaneously with either saline or kainate (35 mg/kg). The animals were observed for 3 h to score convulsions (Rebola et al., 2005a (link)) according to the original Racine scale for rats or the Racine scale adapted to mice and then maintained in groups of three to four per cage.

Canas P.M., Porciúncula L.O., Simões A.P., Augusto E., Silva H.B., Machado N.J., Gonçalves N., Alfaro T.M., Gonçalves F.Q., Araújo I.M., Real J.I., Coelho J.E., Andrade G.M., Almeida R.D., Chen J.F., Köfalvi A., Agostinho P, & Cunha R.A. (2018). Neuronal Adenosine A2A Receptors Are Critical Mediators of Neurodegeneration Triggered by Convulsions. eNeuro, 5(6), ENEURO.0385-18.2018.

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Diverse Pharmacological Interventions in Mice

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DMCM, NMDA, picrotoxin, AMPA, and Kainate were purchased by Tocris (Buckhurst Hill, United Kingdom), bicuculline, and PTZ from Sigma (St. Louis, MO, United States). For systemic injections, all compounds (picrotoxin, PTZ, DMCM, bicuculline) were given intraperitoneally (i.p.) at a volume of 0.1 mL/10 g of mouse body weight. For intracerebroventricular (i.c.v.) injections (NMDA, AMPA, or Kainate), mice were anesthetized with isoflurane, and injections were made in the left or right lateral ventricle (coordinates 1 mm posterior and 1 mm lateral to the bregma; depth 2.4 mm) using a 5-μL Hamilton microsyringe; the injections of the drug by this procedure led to a uniform distribution throughout the ventricular system within 10 min (De Sarro et al., 2004a (link)). All solutions were freshly prepared before the experiments.

Tallarico M., Leo A., Russo E., Citraro R., Palma E, & De Sarro G. (2023). Seizure susceptibility to various convulsant stimuli in the BTBR mouse model of autism spectrum disorders. Frontiers in Pharmacology, 14, 1155729.

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Intracortical Mouse Model for mTLE-HS

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We used the intracortical mouse model for mTLE-HS, described in detail by Bedner et al. (11 (link)), to elicit status epilepticus. In brief, the mice were anesthetized with a mixture of medetomidine (0.3 mg/kg, i.p.) and ketamine (40 mg/kg, i.p.) and kept on a heating blanket. For mice in the kainic acid group, kainate (70 nl, 20 mM, Tocris) was injected above the right hippocampus (= ipsilateral) by a Hamilton pipette (Hamilton Company, NV) at a depth of 1.7 mm at anteroposterior −2 mm, lateral +1.5 mm in relation to bregma. After the procedure, anesthesia was stopped with atipamezole (300 mg/kg, i.p.). Buprenorphine (0.1 mg/kg, s.c.) was applied at 4 and 12 h after the intervention. In order to ensure successful execution of technical procedures, only animals displaying convulsive seizures (Racine grade 5) within the first 4 h after termination of the procedures were included in further analysis. For sham animals, 0.9% NaCl was used instead of kainate for the intracortical injection.

Berger T.C., Vigeland M.D., Hjorthaug H.S., Nome C.G., Taubøll E., Selmer K.K, & Heuser K. (2020). Differential Glial Activation in Early Epileptogenesis—Insights From Cell-Specific Analysis of DNA Methylation and Gene Expression in the Contralateral Hippocampus. Frontiers in Neurology, 11, 573575.

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Pharmacology of Neurotransmitter Signaling

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Salts and general reagents were purchased from Sigma (St. Louis, MO, USA); GYKI 53655, D-AP5, NBQX, bicuculline, Rp-Br-cAMP, H-89, forskolin, philanthotoxin, ryanodine, thapsigargin, kainate, Pertussis toxin, CMZ and W-7 were obtained from Tocris (Bristol, UK).

Falcón-Moya R., Losada-Ruiz P, & Rodríguez-Moreno A. (2019). Kainate Receptor-Mediated Depression of Glutamate Release Involves Protein Kinase A in the Cerebellum. International Journal of Molecular Sciences, 20(17), 4124.

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Acute Slice Excitotoxicity Assay

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Acute slices prepared from P14 Reln^wt mice were allowed to recover after transfection, pharmacological blockers used during the transfection process were thoroughly washed out, and slices were kept under carbogene-gassed ACSF. To induce cell damage, we stimulated non-transfected acute slices with 100 μM AMPA or 25 μM kainate or 50 μM NMDA (all from Tocris, Wiesbaden,Germany) for 15 min. Subsequently, acute slices were incubated with 10 nM propidium iodide (PI; Molecular Probes, Eugene, OR,USA) for 5 min. PI is a non-cell permeant in healthy cells and is employed as membrane permeability marker which emits fluorescence upon binding to nuclear DNA in impaired cells. Afterwards, excess PI was washed out several times with ACSF, and acute slices were mounted on an inverted laser microscope for imaging. Images were acquired at 10 × magnification (512 × 512 pixel resolution). Cell counting was calculated with MacBiophotonis ImageJ software. The number of PI stained cells was counted in each ROI, divided by the average number of PI stained cells from untreated control (non-transfected) acute slices, and plotted as fold increase in death rate.

Hamad M.I., Daoud S., Petrova P., Rabaya O., Jbara A., Melliti N., Stichmann S., Reiss G., Herz J, & Förster E. (2020). Biolistic transfection and expression analysis of acute cortical slices. Journal of neuroscience methods, 337, 108666.

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Adenosine-Mediated Signaling Assay

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N6-cyclopentyladenosine (CPA), adenosine triphosphate (ATP) and malachite green were from Sigma-Aldrich (Madrid, Spain). Ammonium molybdate was from Merck (Madrid, Spain). CGS 21680, NMDA, AMPA and Kainate were from Tocris Bioscience (Madrid, Spain). All other reagents were of analytical grade and obtained from commercial sources.

Crespo M., León-Navarro D.A, & Martín M. (2022). Na+/K+- and Mg2+-ATPases and Their Interaction with AMPA, NMDA and D2 Dopamine Receptors in an Animal Model of Febrile Seizures. International Journal of Molecular Sciences, 23(23), 14638.

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Kainate-Induced Hippocampal Sclerosis Model

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Mice were allowed to recover from surgery for at least 1 week before induction of status
epilepticus, which leads to hippocampal sclerosis and chronic spontaneous seizures after
∼2 weeks.³⁰ (link) Mice were
anaesthetized with isoflurane and were injected with 1 ml of 5% dextrose saline, to
prevent dehydration during status epilepticus. Kainate (100 nl, 20 mM in saline, Tocris)
was infused into the left dorsal hippocampus targeting the molecular layer of the dentate
gyrus, via an injection cannula (internal cannula with 0.2 mm projection for a 1.6 mm
ventral from the brain surface, PlasticsOne), through the previously implanted guide
cannula resulting in status epilepticus. Chronic seizure manifestation was not confirmed
in mice to be utilized solely for anatomical analyses or in experiments to test
functionality of MSGN optical stimulation to entrain hippocampal-wide oscillations.
However, behavioural manifestations of status epilepticus upon Kainate injection and
hippocampal sclerosis had to be present for inclusion in the study.

Hristova K., Martinez-Gonzalez C., Watson T.C., Codadu N.K., Hashemi K., Kind P.C., Nolan M.F, & Gonzalez-Sulser A. (2021). Medial septal GABAergic neurons reduce seizure duration upon optogenetic closed-loop stimulation. Brain, 144(5), 1576-1589.

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Drug Preparation for Seizure Susceptibility

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For seizure susceptibility testing, flurothyl (Bis(2,2,2-trifluoroethyl ether) and pentylenetetrazole (PTZ) were obtained from Sigma-Aldrich, and kainate was obtained from Tocris. All drugs were prepared immediately prior to test injections at their final concentrations and shielded from light. For use in acute slice electrophysiology experiments, R-CPP and NBQX disodium salt were obtained from Tocris Bioscience. Na₃GTP and TTX were from Roche and Alomone Laboratories, respectively. Drugs were dissolved as stock solutions in either water or DMSO and aliquoted and frozen at −30 °C before use. Each drug was diluted to the appropriate concentrations by adding to the perfusate immediately before the experiment. The final concentration of DMSO in the perfusate was 0.1%.

Gertler T.S., Cherian S., DeKeyser J.M., Kearney J.A, & George AL J.r. (2022). KNa1.1 gain-of-function preferentially dampens excitability of murine parvalbumin-positive interneurons. Neurobiology of disease, 168, 105713.

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Glutamate Receptor Agonists and Antagonists Protocol

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RLZ (Merck, Milan, Italy) was dissolved in 10% DMSO and 90% sterile saline. NMDA, AMPA and kainate were purchased from Tocris (Buckhurst Hill, UK). The doses of glutamate receptor agonists (NMDA, AMPA) were dissolved in saline and delivered in a volume of 2 µL by a Hamilton syringe connected to a CMA/100 infusion pump. The competitive NMDA receptors antagonist, (±) 3-(2-carboxypiperazine-4-yl)-1-phosphonate) (CPP), was purchased from Merck and dissolved in normal saline solution to obtain the desired concentrations. The non-competitive AMPA receptor antagonist, N-acetyl-1-(4-chlorophenyl)-6,7-dimethoxy-1,2,3,4-tetrahydroisoquinoline (THIQ-10c), was synthesized by Prof. Alba Chimirri’s group (Department of Medicinal Chemistry, University of Messina, Messina, Italy). THIQ-10c was dissolved in a 40% 2-hydroxypropyl-γ-cyclodextrin with sonication and mild heating.

Citraro R., Bosco F., Di Gennaro G., Tallarico M., Guarnieri L., Gallelli L., Rania V., Siniscalchi A., De Sarro G, & Leo A. (2023). An In Vivo Electroencephalographic Analysis of the Effect of Riluzole against Limbic and Absence Seizure and Comparison with Glutamate Antagonists. Pharmaceutics, 15(7), 2006.

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Repeated, Low-Dose Kainate Epilepsy Model

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A description of the methods for and characteristics of the repeated, low‐dose kainate model have been published in detail elsewhere.9, 10, 37 Briefly, convulsive status epilepticus was induced in adult male Sprague–Dawley rats (180–200 g) with repeated (i.e., approximately every 30 min), low‐dose (7.5 mg/kg), intraperitoneal (IP) injections of kainic acid (Tocris Bioscience, Bristol, UK). During and after the kainate treatment, all rats were maintained in a climate‐controlled vivarium. All of the procedures conformed to the NIH guidelines (i.e., as described in The Guide for Care and Use of Laboratory Animals) and were approved by the University of Utah Animal Care and Use Committee (IACUC).

Berdichevsky Y., Saponjian Y., Park K., Roach B., Pouliot W., Lu K., Swiercz W., Dudek F.E, & Staley K.J. (2016). Staged anticonvulsant screening for chronic epilepsy. Annals of Clinical and Translational Neurology, 3(12), 908-923.

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