Ryanodine

Manufactured by Bio-Techne

Sourced in United Kingdom, United States

Ryanodine is a chemical compound used in research and laboratory settings. It acts as a selective agonist for ryanodine receptors, which are responsible for the release of calcium from intracellular stores. Ryanodine binds to these receptors, modulating their activity and affecting cellular calcium signaling.

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Lab products found in correlation

33 protocols using ryanodine

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 Dynamics in Regenerating Tadpole Tissues

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Regenerating tissues of amputated tadpoles were dissociated with 1.5
mg/ml Proteinase K. Cells from the regenerating tissues were collected from
amputated tadpoles at various time points during 2–73 hours post
amputation (hpa) and plated in saline (in mM: 117 NaCl, 0.7 KCl, 1.3
MgSO₄, 2 CaCl₂, 4.6 Tris, pH 7.8). After 30 min cells
were loaded with 1 μM Fluo4-AM (Life Technologies, Inc.) and imaged with
a Nikon swept-field confocal microscope at an acquisition rate of 0.2 Hz for 1 h
[26 (link), 27 (link)].
To assess the mechanisms mediating Ca²⁺ transients in
regenerating tissues, cells were Ca²⁺-imaged in the presence
of either saline only, vehicle (0.5% DMSO or 0.5% ethanol),
Ca²⁺-free saline or 5–50 μM ryanodine
(Tocris, pre-incubated for 30 min). We performed a paired comparison of
Ca²⁺ activity before and after addition of any agent.

Tu M.K, & Borodinsky L.N. (2014). Spontaneous calcium transients manifest in the regenerating muscle and are necessary for skeletal muscle replenishment. Cell calcium, 56(1), 34-41.

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Preparation of Pharmacological Compounds

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Tetraethylammonium, caffeine, estrogen and progesterone were purchased from MilliporeSigma (St. Louis, MO), and iberiotoxin and ryanodine from Tocris (Minneapolis, MN). Stocks of tetraethylammonium, caffeine and iberiotoxin were prepared in deionized H₂O, whereas stocks of estrogen, progesterone, and ryanodine were prepared in DMSO. Stocks were then diluted to the appropriate concentration in corresponding experimental solutions and used immediately.

Hu X.Q., Song R., Romero M., Dasgupta C., Huang X., Holguin M.A., Williams V., Xiao D., Wilson S.M, & Zhang L. (2019). Pregnancy increases Ca2+ sparks/STOCs and reduces uterine arterial myogenic tone. Hypertension (Dallas, Tex. : 1979), 73(3), 691-702.

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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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Perfusion of Tissues with Oxygenated KRB Solution

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All tissues were perfused continuously with KRB solution containing (mmol/L): NaCl, 5.9; NaHCO₃, 120.35; KCl, 1.2; MgCl₂, 15.5; NaH₂PO₄,1.2; CaCl₂, 2.5; and glucose, 11.5. The KRB solution was warmed to a physiological temperature of 37 ± 0.3°C and bubbled with a mixture of 97% O₂ – 3% CO₂. For experiments utilizing external solutions with 0 [Ca²⁺]_o, CaCl₂ was omitted and 0.5 mM ethylene glycol-bis (β-aminoethyl ether)-N, N, N’, N’–tetraacetic acid (EGTA) was added to the solution. NNC 55–0396 and TTA-A2 were purchased from Alomone Labs (Jerusalem, Israel). 2-aminoethyl-diphenylborinate (2-APB), tetracaine, nicardipine, pinacidil was purchased from Millipore-Sigma (St. Louis, Missouri, USA). Thapsigargin, isradipine, Z-944, CPA and ryanodine were purchased from Tocris Bioscience (Ellisville, Missouri, USA). GSK 7975A was purchased from Aobious (Aobious INC, MA, USA), and xestospongin C (XeC) was purchased from Cayman Chemical (Michigan, USA).

Baker S.A., Leigh W.A., Del Valle G., De Yturriaga I.F., Ward S.M., Cobine C.A., Drumm B.T, & Sanders K.M. (2021). Ca2+ signaling driving pacemaker activity in submucosal interstitial cells of Cajal in the murine colon. eLife, 10, e64099.

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Pharmacological Modulation of Cellular Pathways

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TUDCA (sodium salt), N-acetylcysteine, estrogen, and progesterone were purchased from MilliporeSigma, GSK2606414 and ryanodine from Tocris (Minneapolis, MN), and EUK-134 from Cayman Chemical (Ann Arbor, MI). Stocks of TUDCA and N-acetylcysteine were prepared in deionized H₂O, whereas stocks of estrogen, progesterone, GSK2606414, EUK-134 and ryanodine were prepared in DMSO. Stocks were then diluted to the appropriate concentration in corresponding experimental solutions and used immediately. An equal volume of vehicle was also used to serve as a control.

Hu X.Q., Song R., Romero M., Dasgupta C., Min J., Hatcher D., Xiao D., Blood A., Wilson S.M., Zhang L, & Longo L.D. (2020). Gestational hypoxia inhibits pregnancy-induced upregulation of Ca2+ sparks and spontaneous transient outward currents in uterine arteries via heightened endoplasmic reticulum/oxidative stress. Hypertension (Dallas, Tex. : 1979), 76(3), 930-942.

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Neuromodulators and Calcium Signaling in Retinal Ganglion Cells

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Ex-4, nimodipine, Ex-9-39 and BayK-8644 were purchased from MedChemExpress (Monmouth Junction, NJ, USA), while TTX, Ryanodine, W-7 and U-73122 were from Tocris Bioscience (Ellisville, MO, USA). NF-449 was purchased from Cayman Chemical (Ann Arbor, MI, USA) and C2 Ceramide from Santa Cruz (Dallas, TX, USA). The others were purchased from Sigma-Aldrich. nimodipine, Bis-IV, U-73122, Ryanodine, W-7, FK-506, OA, and C2 Ceramide were initially dissolved in DMSO for stock. The final concentration of DMSO was less than 0.1%, with no effects on the calcium currents of RGCs. All other drug solutions were prepared in ion-free water, stored at −20°C and freshly diluted to the final concentrations using extracellular or intracellular solutions. For topical ocular administration, Ex-4 and Ex-9-39 were dissolved in saline, and BayK-8644 was dissolved in 50% PEG-400 diluted with saline.

Wang Y.C., Wang L., Shao Y.Q., Weng S.J., Yang X.L, & Zhong Y.M. (2023). Exendin-4 promotes retinal ganglion cell survival and function by inhibiting calcium channels in experimental diabetes. iScience, 26(9), 107680.

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Modulating Intracellular Calcium Signaling

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Pertussis toxin (PTX, 200 ng/mL; Sigma), an ADP ribosylating agent, was used to specifically inactivate G_i/o class G proteins. WU‐07047 (20 μmol/L; Washington University) was used to inhibit G_q class G proteins. Thapsigargin (1 μmol/L; Tocris Biosciences) was used to block sarcoplasmic reticulum (SR) Ca²⁺ uptake via SERCA pump. Ryanodine (10 μmol/L; Tocris Biosciences) was used to target Ryanodine receptors (RyRs). N,N,N′,N′‐tetrakis(2‐pyridylmethyl)ethane‐1,2‐diamine (TPEN, 50 μmol/L; Sigma) was used to sequester Ca²⁺ in intracellular stores. Ionomycin (5 μmol/L) was used to stimulate Ca²⁺ release from internal stores.

Jie L., Owens E.A., Plante L.A., Fang Z., Rensing D.T., Moeller K.D, & Osei‐Owusu P. (2016). RGS2 squelches vascular Gi/o and Gq signaling to modulate myogenic tone and promote uterine blood flow. Physiological Reports, 4(2), e12692.

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Standardized Pharmacological Stimulation for Tissue and Organoid Imaging

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Stimulant stock solutions were prepared according to
manufacturer’s instructions. For mucus measurement and tissue imaging
experiments 10 μM Cch (Sigma Aldrich) was provided basolaterally.
Stimulants were added to tissue culture medium for organoid imaging experiment
at the following concentrations: acetylcholine (20 μM; Sigma Aldrich),
Cch (20 μM; Sigma Aldrich), forskolin (100 nM; Cayman Chemical) and
nicotine (10 μM; Sigma Aldrich).
Inhibitor stock solutions were prepared according to
manufacturer’s instructions. For mucus measurement and tissue imaging
experiments 4-DAMP (100 nM; Cayman Chemical) was supplied basolaterally. For
organoid imaging experiments inhibitors were added to tissue culture medium and
incubated with cells for 10 minutes prior to stimulation with Cch. Inhibitors
were used at the following concentrations; atropine (50 μM; Sigma
Aldrich), physostigmine salicylate (10 μM; Sigma Aldrich), tetraisopropyl
pyrophosphoramide (10 μM; Sigma Aldrich), 4-DAMP (100 nM),
(−)-Xestospongin C (10 μM; Tocris), SKF 96365 (10 μM;
Tocris), carbenoxolone (50 μM; Sigma Aldrich), bumetanide (50μM;
Sigma Aldrich), clotrimazole (30 μM; Sigma Aldrich), Chromanol 293B (10
μM; Sigma Aldrich) and Ryanodine (100 nM; Tocris).

Dolan B., Ermund A., Martinez-Abad B., Johansson M.E, & Hansson G.C. (2022). Clearance of small intestinal crypts involves goblet cell mucus secretion by intracellular granule rupture and enterocyte ion transport. Science signaling, 15(752), eabl5848.

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Pharmacological Agents for Neuroscience Research

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(RS)-3,5-DHPG, LY367385, MPEP, APV, picrotoxin, ryanodine, riluzole were purchased from Tocris Bioscience. Choline chloride was from Junsei Chemical (Tokyo, Japan). 8-NH₂-cADPR was from Molecular Probes. All other drugs were purchased from Sigma-Aldrich (St Louis, MO, USA). Stock solutions of drugs were made by dissolving in deionized water or DMSO according to manufacturer’s specifications and were stored at − 20 °C. On the day of the experiments, one aliquot was thawed and used. The concentration of DMSO in solutions was maintained at 0.1%.

Yu W., Sohn J.W., Kwon J., Lee S.H., Kim S, & Ho W.K. (2018). Enhancement of dendritic persistent Na+ currents by mGluR5 leads to an advancement of spike timing with an increase in temporal precision. Molecular Brain, 11, 67.

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