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8 pcpt cgmp

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8-pCPT-cGMP is a synthetic analogue of the cellular signaling molecule cyclic guanosine monophosphate (cGMP). It functions as a selective agonist for cGMP-dependent protein kinases.

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Lab products found in correlation

Tubocurarine, by Merck Group (1 mentions) Deta no, by Merck Group (1 mentions) 18β ga, by Merck Group (1 mentions) Nω nitro l arginine methyl ester hydrochloride l name, by Merck Group (1 mentions) Carbachol, by Merck Group (1 mentions) 1h 1 2 4 oxadiazolo 4 3 a quinoxalin 1 one, by Merck Group (1 mentions) Fluo 4 acetoxymethyl ester, by Thermo Fisher Scientific (1 mentions) S nitroso n acetyl dl penicillamine snap, by Merck Group (1 mentions) Carbachol, by Thermo Fisher Scientific (1 mentions) Caffeine, by Merck Group (1 mentions) Zaprinast, by Merck Group (1 mentions) Incucyte, by Sartorius (1 mentions) Mineral oil, by Merck Group (1 mentions) 8 br camp, by Merck Group (1 mentions) Transgenic dough diet, by Bio-Serv (1 mentions) Forskolin, by Merck Group (1 mentions)

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PCPT-cGMP (2 citations)

5 protocols using 8 pcpt cgmp

1

Pharmacological Modulation of Signaling Pathways

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During this study the following drugs were used: diethylenetriamine/nitric oxide adduct (DETA-NO, 250–500 µM, Sigma), Nω-Nitro-L-arginine methyl ester hydrochloride (L-NAME, 500 µM-1 mM, Sigma), 8-(4-Chlorophenylthio)-guanosine 3′, 5′-cyclic monophosphate sodium salt (8-pCPT-cGMP, 500–750 µM, Sigma), 1 H-[1] (link), [2] (link), [4] (link)Oxadiazolo[4,3-a]quinoxalin-1-one (ODQ, 500 µM, Ascent Scientific), tetrodotoxin (TTX, 1 µM, Ascent Scientific), (+)-tubocurarine hydrochloride (tubocurarine, 3 µM, Sigma), formamide (2 M, Sigma) and 18-β-glycyrrhetinic acid (18βGA, 100 µM, Sigma).
Jay M., Bradley S, & McDearmid J.R. (2014). Effects of Nitric Oxide on Neuromuscular Properties of Developing Zebrafish Embryos. PLoS ONE, 9(1), e86930.
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2

Vasopressin-Induced Ca2+ Signaling Regulation

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Physiological saline of the following composition was used (mM): NaCl 120.4, KCl 5.9, MgSO4 1.2, CaCl2 2, glucose 11.5, and HEPES 11. The Ca2+-free solutions contained 2 mM EGTA. [Arg8]-Vasopressin acetate salt (AVP), Carbachol (CCh), Caffeine, Zaprinast (ZAP), 1H-[1,2,4]Oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), Rp-8-pCPT-cGMPS sodium salt, 8pCPT-cGMP, S-nitroso-N-acetyl-DL-penicillamine (SNAP) were from Sigma. Fluo-4 acetoxymethyl ester was from Molecular Probes, Life Technologies, UK.
[Arg8]-Vasopressin acetate salt, Carbachol, Caffeine, Rp-8-pCPT-cGMPS sodium salt, 8pCPT-cGMP were dissolved in water; S-nitroso-N-acetyl-DL-penicillamine (SNAP), 1H-[1,2,4]Oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), Zaprinast in DMSO.
Borysova L, & Burdyga T. (2015). Evidence that NO/cGMP/PKG signalling cascade mediates endothelium dependent inhibition of IP3R mediated Ca2+ oscillations in myocytes and pericytes of ureteric microvascular network in situ. Cell Calcium, 58(6), 535-540.
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3

Apoptosis Monitoring in Renal Proximal Tubular Cells

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Human primary renal proximal tubular epithelial cells (HRPTC, ScienCell Research Laboratories, Carlsbad, CA) were seeded in 96-well plates and cultured for 1 day until 70–80% confluency before experiments. Passages 3–5 were used. Caspase 3/7 green apoptosis assay agent (Essen BioScience, Ann Arbor, MI) was added to culture medium for the detection of apoptosis. Plates were then transferred to a time-lapsed, live imaging system IncuCyte (Essen BioScience, Ann Arbor, MI) for apoptosis monitoring for a total of 24 hours. Hypoxia/reoxygenation (H/R) in cells was mimicked with mineral oil (Sigma, St Louis, MO) immersion 25 (link). Cells were immersed with oil for 2 hours, washed twice and replaced with fresh medium including vehicle (H/R group) or drugs. ANP, BNP, CRRL269 at concentrations of 10−8, 10−7 or 10−6 M were used. Protein kinase G (PKG, important cGMP binding protein) activating analogue 8-pCPT-cGMP (specific for PKG and low affinity to phosphodiesterases, Sigma, St Louis, MO) at concentrations of 10−7, 10−6 or 10−5 M were used. Cells were treated with 10−6, 5X10−6 or 1.5X10−5 M PKG inhibitor KT-5823 (Abcam, Cambridge, MA) for 30 min before 10−6 M CRRL269 was added. Cells without H/R stimulation were labeled as negative control (Neg Ctrl) group. Apoptosis signals (e.g. apoptosis object counts) were analyzed with the software provided by IncuCyte.
Chen Y., Harty G.J., Zheng Y., Iyer S.R., Sugihara S., Sangaralingham S.J., Ichiki T., Grande J.P., Lee H.C., Wang X.L, & Burnett JC J.r. (2019). CRRL269: A Novel Particulate Guanylyl Cyclase A Receptor Peptide Activator For Acute Kidney Injury. Circulation research, 124(10), 1462-1472.
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4

Selective Dual TRPC3/TRPC6 Inhibitor

Cited 1 time
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A new selective dual TRPC3/TRPC6 small molecule inhibitor GSK2833503A (GSK503A, also denoted as Example 19) 25 (link), 26 was provided by GlaxoSmithKline Pharmaceuticals. GSK503A has an IC50 = 21 nM for TRPC3 and 3 nM for TRPC6. Corresponding IC50 for Cav1.2, hERG, Nav1.5, TRPV1, and TRPV4 are 10,000, >50,000, 3300, 6,300, and 12,500 respectively. Cell permeable cGMP-analog (8-pCPT-cGMP) and cAMP-analog (8-Br-cAMP) were obtained from Sigma. For in vivo studies, sildenafil citrate (Revatio, Pfizer) was compressed into soft rodent chow (Transgenic Dough Diet, Bio-Serv) and provided at a dose of 200mg/kg/day for 2 months as described previously 27 (link). This dose yields a free plasma concentration in the range of 30–50 nM, well within the selective range for PDE5A.
Seo K., Rainer P.P., Lee D.I., Hao S., Bedja D., Birnbaumer L., Cingolani O.H, & Kass D.A. (2014). Hyperactive Adverse Mechanical-Stress Responses in Dystrophic Heart are Coupled to TRPC6 and Blocked by cGMP-PKG Modulation. Circulation research, 114(5), 823-832.
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5

Murine Intestinal Tissue and Ussing Chamber Assay

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Collection of murine intestinal tissue and Ussing chamber assays were performed as described elsewhere (41 (link)). Tissue was incubated in modified Meyler solution (128 mmol/liter NaCl, 4.7 mmol/liter KCl, 1.3 mmol/liter CaCl2, 1.0 mmol/liter MgCl2, 0.3 mmol/liter Na2HPO4, 0.4 mmol/liter NaH2PO4, 20 mmol/liter NaHCO3, 10 mmol/liter HEPES), supplemented with glucose (10 mmol/liter; added solely to the serosal bathing solution) in 95% O2, 5% CO2, pH 7.3, at 37 °C. CFTR-dependent anion secretion was stimulated by addition of the adenylyl cyclase activator forskolin (Sigma-Aldrich), STa (added to the luminal bathing solution only; Bachem), or the membrane-permeable cGMP analog 8-pCPT-cGMP (Sigma-Aldrich). Data shown represent Isc measurements, obtained by clamping of the transepithelial potential difference at 0 mV.
Bijvelds M.J., Tresadern G., Hellemans A., Smans K., Nieuwenhuijze N.D., Meijsen K.F., Bongartz J.P., Ver Donck L., de Jonge H.R., Schuurkes J.A, & De Maeyer J.H. (2018). Selective inhibition of intestinal guanosine 3′,5′-cyclic monophosphate signaling by small-molecule protein kinase inhibitors. The Journal of Biological Chemistry, 293(21), 8173-8181.
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