Cyclopiazonic acid

Manufactured by Bio-Techne

Sourced in United Kingdom, United States

Cyclopiazonic acid is a laboratory reagent used for research purposes. It is a mycotoxin produced by several species of fungi. Cyclopiazonic acid functions as a potent inhibitor of calcium-ATPase, an enzyme responsible for the removal of calcium ions from the cytoplasm of cells.

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Lab products found in correlation

Gsk2193874, by Bio-Techne (6 mentions) Gsk1016790a, by Bio-Techne (6 mentions) Fluo 4 am, by Thermo Fisher Scientific (5 mentions) Apyrase, by Merck Group (5 mentions) Ns309, by Bio-Techne (4 mentions) Fura 2 am, by Thermo Fisher Scientific (4 mentions) Tamoxifen, by Merck Group (4 mentions) Ar c 118925xx, by Bio-Techne (4 mentions) Apamin, by Bio-Techne (3 mentions) U73122, by Bio-Techne (3 mentions) Mrs2179, by Bio-Techne (3 mentions) U46619, by Cayman Chemical (3 mentions) Ryanodine, by Bio-Techne (2 mentions) Caffeine, by Merck Group (2 mentions) H1009, by Merck Group (2 mentions) Thapsigargin, by Bio-Techne (2 mentions) Tram 34, by Bio-Techne (2 mentions) Verapamil, by Bio-Techne (2 mentions) Glibenclamide, by Bio-Techne (2 mentions) Ruthenium red, by Bio-Techne (2 mentions)

Spelling variants of this product

Cyclopiazonic acid (CPA) (13 citations)

18 protocols using cyclopiazonic acid

Evaluating PCSK9 and CD36 Regulation in Liver Cell Lines

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HuH7 (kind gift from Dr. Nabil G. Seidah) and HepG2 (ATCC; HB-8065) cells were routinely grown in complete Dulbecco’s Modified Eagle’s Medium (Gibco, Thermofisher Scientific) supplemented with 10% fetal bovine serum (Sigma-Aldrich) and 100 U/ml of penicillin and streptomycin (Sigma-Aldrich). CF, ryanodine, 2 APB, CDN, theobromine, paraxanthine, 8-cyclopentyl-1,3-dimethylxanthine (8CD), 8-(3-Chlorostyryl) CF (8CC), PSB603, cyclopiazonic acid and U18666A were purchased from Tocris Bioscience. All cell treatments were carried out for 24 h unless otherwise stated. Cells were transfected with a cocktail consisting of plasmid DNA (1 µg), X-tremeGENE HP (3 µl; Thermo Fisher Scientific), and opti-MEM (100 µl; Thermo Fisher Scientific) per 1 ml complete medium containing plated cells. Human PCSK9 was overexpressed using the bicistronic pIRES-EGFP plasmid; calnexin using the mPA-GFP-N1 plasmid. To attenuate the expression of GRP78 and CD36, siGENOME smartpool siRNA was purchased from GE Dharmacon (M-008198-02 and L-010206-00-0005 respectively) and transfected using lipofectamine RNAiMAX as per the manufacturer’s protocol.

Lebeau P.F., Byun J.H., Platko K., Saliba P., Sguazzin M., MacDonald M.E., Paré G., Steinberg G.R., Janssen L.J., Igdoura S.A., Tarnopolsky M.A., Wayne Chen S.R., Seidah N.G., Magolan J, & Austin R.C. (2022). Caffeine blocks SREBP2-induced hepatic PCSK9 expression to enhance LDLR-mediated cholesterol clearance. Nature Communications, 13, 770.

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Calcium Signaling Pathway Modulation

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Ionomycin was purchased from Adipogen; Cyclopiazonic Acid (CPA) was obtained from Tocris; Pluronic-F12 from Biotium and Life Technologies; Fluo4-AM from Life Technologies; BTP-2 from EMD-Millipore; 4,4'-diisothiocyano-2,2'-stilbenedisulfonic acid (DIDS) from Sigma.

Sharma A., Ramena G., Yin Y., Premkumar L, & Elble R.C. (2018). CLCA2 is a positive regulator of store-operated calcium entry and TMEM16A. PLoS ONE, 13(5), e0196512.

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Neurochemical Compounds for Functional Assays

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Apamin, iberiotoxin, cyclopiazonic acid, DHPG, DNQX, picrotoxin, quinpirole, SKF81297, SCH23390, CGS21680, and SCH58261 were obtained from Tocris Biosciences. TTX was obtained from Alomone Labs. Fluo-4FF and Alexa Fluor 594 were purchased from Life Technologies. Caged IP₃ was a generous gift from Dr. Kamran Khodakhah at Albert Einstein College of Medicine. All other chemicals were from Sigma-RBI.

Swapna I., Bondy B, & Morikawa H. (2016). Differential Dopamine Regulation of Ca2+ Signaling and Its Timing Dependence in the Nucleus Accumbens. Cell reports, 15(3), 563-573.

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Patch-Clamp Evaluation of Cardiac Ion Channels

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R_N, RMP, RS, and AP output were assessed using whole-cell patch-clamp physiology before and after bath-applied drugs, which involved perfusion through the recording chamber for 10 min followed by wash out for 15 min. These same parameters were examined using drugs dissolved in the internal pipette solution, at time = 0′ and time = 25′. Bath applied drugs were the RyR blocker dantrolene (10 μM; Tocris), the SERCA pump inhibitor cyclopiazonic acid (20 μM; Tocris), the RyR agonist caffeine (10 mM; Sigma-Aldrich), the RyR blocker and beta and alpha adrenergic blocker carvedilol (10 μM; M. Fill), and the partial RyR blocker ryanodol (10 μM). Drugs applied intracellularly via the patch pipette were the IP₃R blocker heparin (1mg/ml) and the SERCA pump inhibitor thapsigargin (2 mM; Calbiochem).

Musial T.F., Molina-Campos E., Bean L.A., Ybarra N., Borenstein R., Russo M.L., Buss E.W., Justus D., Neuman K.M., Ayala G.D., Mullen S.A., Voskobiynyk Y., Tulisiak C.T., Fels J.A., Corbett N.J., Carballo G., Kennedy C.D., Popovic J., Ramos-Franco J., Fill M., Pergande M.R., Borgia J.A., Corbett G.T., Pahan K., Han Y., Chetkovich D.M., Vassar R.J., Byrne R.W., Oh M.M., Stoub T.R., Remy S., Disterhoft J.F, & Nicholson D.A. (2018). Store depletion-induced h-channel plasticity rescues a channelopathy linked to Alzheimer’s disease. Neurobiology of learning and memory, 154, 141-157.

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Oxidative Stress Assay Reagent Preparation

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H₂O₂ working solutions were freshly prepared before every experiment from H₂O₂ solution 30 % (w/w) in H₂O (Sigma-Aldrich, H1009). Stock solution of Menadione (VENDOR, CAT) was prepared in 100% DMSO at 50mM. Stock solution of Cyclopiazonic Acid (Tocris, 1235) was prepared in 100% DMSO at 20mM. Chemicals specific to other method sections can be found in their respective sections.

Berndt A., Lee J., Won W., Kimball K., Neiswanger C., Schattauer S., Wang Y., Yeboah F., Ruiz M., Evitts K., Rappleye M., Bremner S., Chun C., Smith N., Mack D., Young J., Lee C.J, & Chavkin C. (2024). Ultra-fast genetically encoded sensor for precise real-time monitoring of physiological and pathophysiological peroxide dynamics. Research Square.

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Purinergic Autoreceptor Modulation in Astrocytes

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Drugs were purchased from either Sigma (apyrase, thapsigargin, cyclopiazonic acid – CPA) or Tocris Bioscience (MRS 2179, SQ22536 and U73122). Two approaches were used to investigate involvement of purinergic autoreceptors on astrocytes. MRS 2179 is widely considered to be a selective antagonist of metabotropic P2Y1 receptors, but may also affect P2X1 and P2X3 receptors [21] (link). apyrase is an ATP-degrading enzyme which can be used to non-discriminately block the actions of extracellular ATP. apyrase was applied for 30 min before light-stimulation of astrocytes.

Figueiredo M., Lane S., Stout RF J.r., Liu B., Parpura V., Teschemacher A.G, & Kasparov S. (2014). Comparative analysis of optogenetic actuators in cultured astrocytes. Cell Calcium, 56(3), 208-214.

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Comprehensive Calcium Signaling Assay

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Apamin, cyclopiazonic acid (CPA), GSK2193874, GSK1016790A, HC067047, NS309, 1400W, Nω‐propyl‐l‐arginine hydrochloride, ODQ, RN1747, Rp‐8‐Br‐PET‐cGMPS, Suramin, and Tram 34 were purchased from Tocris Bioscience (Minneapolis, MN). ATP and N^ω‐nitro‐l‐arginine were obtained from Sigma‐Aldrich (St. Louis, MO). DAF‐FM diacetate, Fluo‐4AM (Ca²⁺ indicator), and EGTA‐AM (Ca²⁺ chelator) were purchased from Fischer Scientific (Pittsburgh, PA). Spermine NONOate and U46619 were purchased from Cayman Chemical (Ann Arbor, MI). All other chemicals were obtained from Sigma‐Aldrich (St. Louis, MO).

Marziano C., Hong K., Cope E.L., Kotlikoff M.I., Isakson B.E, & Sonkusare S.K. (2017). Nitric Oxide–Dependent Feedback Loop Regulates Transient Receptor Potential Vanilloid 4 (TRPV4) Channel Cooperativity and Endothelial Function in Small Pulmonary Arteries. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 6(12), e007157.

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Vasodilation Mechanisms Investigated

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Sodium pentobarbital was obtained from Sanofi (Brussels, Belgium), indomethacin from CERTA (Belgium), N^Ω-nitro-L-arginine methyl ester (L-NAME), N^Ω-nitro-L-arginine (L-NNA) and nifedipine from Sigma (Bornem, Belgium), Fura 2-AM from Molecular Probes (Invitrogen, Merelbeke, Belgium), 2-aminoethoxydiphenyl borate (2-APB), BAY K8644, levcromakalim, glibenclamide, cyclopiazonic acid (CPA), diltiazem, verapamil from TOCRIS (Bristol, United Kingdom).

Fransen P., Van Hove C.E., Leloup A.J., Martinet W., De Meyer G.R., Lemmens K., Bult H, & Schrijvers D.M. (2015). Dissecting out the Complex Ca2+-Mediated Phenylephrine-Induced Contractions of Mouse Aortic Segments. PLoS ONE, 10(3), e0121634.

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Calcium Signaling Imaging with Fura-2

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Fura-2 pentapotassium salt, Fura-2/AM, and Pluronic F-127 (20% solution in DMSO) were purchased from Life Technologies (Grand Island, NY). Verapamil and cyclopiazonic acid (CPA) were obtained from Tocris Bioscience (Minneapolis, MN). Other chemicals were from Sigma-Aldrich (St. Louis, MO). During experiments, cells were continually perfused at 0.5 ml/min (or 10 ml/min for faster drug delivery) with a physiological solution containing (in mM): 140 NaCl, 5.4 KCl, 1.5 MgCl₂, 2 CaCl₂, 10 glucose, and 10 HEPES (pH 7.3, 300–310 mOsm/kg). For Ca²⁺-free conditions, CaCl₂ was replaced with 2 mM Na-EGTA.

Semenov I., Zemlin C., Pakhomova O.N., Xiao S, & Pakhomov A.G. (2015). Diffuse, non-polar electropermeabilization and reduced propidium uptake distinguish the effect of nanosecond electric pulses. Biochimica et biophysica acta, 1848(10 0 0), 2118-2125.

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Assessing Calcium Signaling Pathways

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The complete list of antibodies used is provided in Table S1. N-benzyl-p-toluene sulfonamide (BTS), thapsigargin, cyclopiazonic acid (CPA), and colchicine were purchased from Tocris Biosciences (Minneapolis, Mn, USA). Proteostat aggresome detection kit (ENZ-51035-K100) and MG132 were purchased from Enzo Life Sciences (Farmingdale, NY, USA). The Ca²⁺ indicators Mag Fluo-4 AM, Fura-2 AM, Halt protease and phosphatase inhibitor cocktail, ER-Tracker Blue-White DPX, TRIzol reagent and laminin were purchased from Thermo Fisher (Waltham, MA, USA). iScript cDNA Synthesis Kit, iQ SYBR Green Supermix, precast and acrylamide gel kits containing Stain Free reagent, molecular weight markers and Starbright secondary antibodies were purchased from Bio-Rad (Hercules, CA, USA). All other chemicals were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Hanna A.D., Lee C.S., Babcock L., Wang H., Recio J, & Hamilton S.L. (2021). Pathological mechanisms of vacuolar aggregate myopathy arising from a Casq1 mutation. FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 35(5), e21349.

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