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Endothelin 1

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Endothelin-1 is a laboratory product manufactured by Merck Group. It is a peptide that functions as a potent vasoconstrictor, playing a role in the regulation of vascular tone and blood pressure.

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Lab products found in correlation

Phenylephrine, by Merck Group (5 mentions) Ns309, by Merck Group (4 mentions) Bq123, by Merck Group (4 mentions) Angiotensin 2, by Merck Group (3 mentions) Histamine, by Merck Group (3 mentions) Bradykinin, by Merck Group (3 mentions) L name, by Merck Group (3 mentions) Indomethacin, by Merck Group (3 mentions) Acetylcholine, by Merck Group (3 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions) Chloroquine, by Merck Group (2 mentions) Platelet activator factor, by Merck Group (2 mentions) Rantes, by Merck Group (2 mentions) Bovine serum albumin (bsa), by Roche (2 mentions) Apamin, by Merck Group (2 mentions) Substance p, by Merck Group (2 mentions) Tram 34, by Merck Group (2 mentions) Adenosine diphosphate (adp), by Merck Group (2 mentions) Norepinephrine, by Merck Group (2 mentions) Apocynin, by Merck Group (2 mentions)

52 protocols using endothelin 1

1

Endothelin-1 Induced Scratching Behavior

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50 μl of vehicle (0.9% saline, Fresenius Kabi) or endothelin-1 (20 pmol, Sigma-Aldrich) were injected intradermally in the back of the neck. Where the ETAR antagonist BQ-123 was used, 10 pmol of endothelin-1 were injected, with or without 10 nmol of BQ-123 (Sigma-Aldrich) in one intradermal injection. Each mouse was then placed in a cage with wooden chips and recorded for 1 h using a digital video camera. Afterwards, AniTracker® v1.2 was used to score the videos for the duration and frequency of grooming and scratching behavior. A scratching episode was defined as a bout of scratching by either hind paw; from the time point it was lifted until placed back on the ground. The videos were scored by observers blinded to the genotype and treatment given. The results for each group are expressed as the mean frequency of scratching episodes/60 min, the mean duration of scratching episodes/60 min, and, where relevant, as the mean length of scratching episodes (total duration/total frequency) and as the mean frequency of scratching episodes per each 10 min interval. Results are presented as mean ± SEM.
Magnúsdóttir E.I., Grujic M., Bergman J., Pejler G, & Lagerström M.C. (2020). Mouse connective tissue mast cell proteases tryptase and carboxypeptidase A3 play protective roles in itch induced by endothelin-1. Journal of Neuroinflammation, 17, 123.
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2

Intradermal Endothelin-1 and Intravenous Oxaliplatin/Octoxynol-9 Protocol

Cited 3 times
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The following drugs and reagents were used in this study: endothelin-1, oxaliplatin, and octoxynol-9, all obtained from Sigma-Aldrich (St. Louis, MO, USA). endothelin-1 (100 ng) was dissolved in saline and administered intradermally on the dorsum of the hind paw, in a volume of 5 μl, using a 30-gauge hypodermic needle attached to a microsyringe (10 μl; Hamilton, Reno, NV, USA) by a short length of polyethylene tubing (PE-10). oxaliplatin (dissolved in saline to a concentration of 2 mg/kg), used to induce painful peripheral neuropathy [16 (link), 28 (link), 32 (link)], and octoxynol-9 (0.5 % solution in saline, at a volume of 1 ml/kg), used to evaluate the contribution of the endothelium to nociceptive mechanisms [4 (link), 13 (link), 14 (link)], were administered intravenously via tail vein injection, followed by a bolus injection of an equal volume of saline before removal of the injection needle. Drug doses were selected based on the results of our previous studies [3 (link), 4 (link), 16 (link), 33 (link)].
Ferrari L.F., Bogen O., Green P, & Levine J.D. (2015). Contribution of Piezo2 to endothelium-dependent pain. Molecular Pain, 11, 65.
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3

Hypertrophic Cardiomyocyte Characterization

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HL-1 cells were grown in Claycomb medium without norepinephrine for 14 days to induce their responsibility to hypertrophic agents according to.23 (link) 3 × 104 HL-1 cells were seeded in 24-well plates 24 hours prior to the experiments and exposed to samples and positive controls (100 nM endothelin 1 and 10 µM norepinephrine, Sigma Aldrich) for 18 hours. Medium-exposed cells served as negative control. CellMask deep red plasma membrane stain (Life Techologies, Poggersdorf, Austria) and Hoechst 33342 were added for 10 minutes at 37°C, and cells subsequently fixed with 4% paraformaldehyde for 10 minutes. Staining was performed according to the manual of the manufacturer. Analysis was performed with Nikon NIS Elements software.29 (link)Cells were viewed at confocal high content screening system (Nikon CEE GmbH, Austria) with 395 nm/414 to 450 nm for Hoechst 33342 and 640 nm/660 to 850 nm for CellMask. Images were analyzed using NIS Elements software with JOBS module. Experiments were performed in triplicates, and 4 fields of view (= 1 827 038 µm2) were analyzed per sample. Cellular area/cell was measured.
Panov V., Minigalieva I., Bushueva T., Fröhlich E., Meindl C., Absenger-Novak M., Shur V., Shishkina E., Gurvich V., Privalova L, & Katsnelson B.A. (2020). Some Peculiarities in the Dose Dependence of Separate and Combined In Vitro Cardiotoxicity Effects Induced by CdS and PbS Nanoparticles With Special Attention to Hormesis Manifestations. Dose-Response, 18(1), 1559325820914180.
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4

Endothelin-1 Signaling Pathway Analysis

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Chemicals that were used in this study: Endothelin-1, BQ123, Tetramethylrhodamine B isothiocyanate-conjugated Phalloidin, and Latrunculin B were purchased from Sigma-Aldrich. GSK429286 and Y27632 were obtained from Selleck-chem. C3 was purchased from Cytoskeleton Inc. Phos-tag-conjugated acrylamide was purchased from Wako Chemicals. DAPI was purchased from Invitrogen.
Wang Z., Liu P., Zhou X., Wang T., Feng X., Sun Y.P., Xiong Y., Yuan H.X, & Guan K.L. (2017). Endothelin Promotes Colorectal Tumorigenesis by Activating YAP/TAZ. Cancer research, 77(9), 2413-2423.
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5

NFAT-luciferase Reporter Cardiomyocyte Assay

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NFAT-luciferase mice _ENREF_99 (link), kindly provided by Dr. Jeffery D. Molkentin (Cincinnati Children’s Hospital Medical Center, Cincinnati, OH), carry nine copies of NFAT-binding sites from the interleukin-4 promoter, upstream of the luciferase gene. Primary ventricular cardiomyocyte cultures from neonatal (one-three days old) NFAT-luciferase reporter mice were prepared as described54 (link). Cell cultures from three separate isolations were used for experiments. Cardiomyocytes were maintained in serum-free medium 24 hours prior to treatment with recombinant Wnt5a, sFRP3 (R&D Systems, Minneapolis, MN, USA) or endothelin-1 (used as positive control, Sigma-Aldrich, St. Louis, MO, USA) for 24 hours, washed twice with PBS and harvested for luciferase activity quantification according to the luciferase assay protocol (Promega, Madison, WI, USA). Luminescence from duplicates was quantified on a Victor 3 1420 Multilabel Counter (PerkinElmer, MA). Cell culture medium was collected for cell viability analyses (ToxiLight, Lonza Group Ltd, Basel, Switzerland) and measurements of sFRP3 release measured by EIAs (R&D Systems; Stillwater, MN, USA).
Abraityte A., Lunde I.G., Askevold E.T., Michelsen A.E., Christensen G., Aukrust P., Yndestad A., Fiane A., Andreassen A., Aakhus S., Dahl C.P., Gullestad L., Broch K, & Ueland T. (2017). Wnt5a is associated with right ventricular dysfunction and adverse outcome in dilated cardiomyopathy. Scientific Reports, 7, 3490.
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6

Vasoactive Contraction of Variable-Modulus Tissue Films

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The construct was placed in 37°C, 1X, pH 7.4 Tyrode’s buffer and cut eight times parallel to tissue alignment and two orthogonal to the initial eight (Hald et al., 2015 ), leaving two rows of four, semi-detached, curved variable-modulus vMTFs with the VSMCs on the concave surface (Fig. 2B). After 15min of equilibration, the variable-modulus vMTF VSMCs were serially stimulated with increasing concentrations of endothelin-1 (Sigma Aldrich) every 10min; 500pM, 5nM, 50nM, and 500nM. Finally, 100μM rho-kinase inhibitor HA-1077 (Sigma Aldrich) was added for 30min. Fluorescent and bright field images were acquired every 30s using a Lumar V12 stereomicroscope (Zeiss, Oberkochen, Germany). These images were used to determine the 2D projection length of the film, which was used to evaluate its radius of curvature (Win et al., 2014 (link)). Data was analyzed using ANOVA and pairwise comparisons were made using the Tukey test in MATLAB.
Steucke K.E., Tracy P.V., Hald E.S., Hall J.L, & Alford P.W. (2015). Vascular smooth muscle cell functional contractility depends on extracellular mechanical properties. Journal of biomechanics, 48(12), 3044-3051.
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7

Endothelin-1 and NS309 Protocols

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Endothelin-1 and NS309 were purchased from Sigma-Aldrich (St. Louis, MO).
Liu Y., Cole V., Lawandy I., Ehsan A., Sellke F.W, & Feng J. (2018). Decreased coronary arteriolar response to KCa channel opener after cardioplegic arrest in diabetic patients. Molecular and Cellular Biochemistry, 445(1), 187-194.
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8

Molecular Signaling Pathways Exploration

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Phenylephrine (Phe, α receptor agonist), acetylcholine, endothelin 1 (ET-1, endothelin receptor agonist), adenosine 5′-triphosphate disodium salt (ATP-Na2), 2-aminoethyl diphenylborinate (2APB, inositol triphosphate receptor inhibitor) were purchased from Sigma and dissolved in the distilled water. Thapsigargin (TG) was obtained from Calbiochem and dissolved in dimethyl sulfoxide (DMSO). The primary goat (sc-10,377) and rabbit (sc-25,749) antibodies against TRPP2, the primary rabbit antibody against IP3 receptor and the primary rabbit antibody against Orai1 were purchased from Santa Cruz Biotechnology. The primary rabbit antibody against STIM1 was obtained from ProSci. Fluo-4 fluorescence dye, TRPP2 small interfering RNA (siRNA), STIM1 siRNA, RNAiMax reagent, lipofectamine 2000, goat anti-rabbit IgG conjugated to Alexa Fluor 488 were purchased from Invitrogen. Protein A magnetic bead was obtained from Millipore.
Guo J., Zhao R., Zhou M., Li J., Yao X., Du J., Chen J, & Shen B. (2020). TRPP2 and STIM1 form a microdomain to regulate store-operated Ca2+ entry and blood vessel tone. Cell Communication and Signaling : CCS, 18, 138.
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9

Immunofluorescence Staining of Pluripotency and Cardiac Markers

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Immunofluorescence staining for pluripotency of iPSCs was performed by using the following primary antibodies: anti‐NANOG (Abcam), anti‐OCT3/4 (Santa Cruz), anti–SSEA 3 (Millipore), anti–SSEA 4 (Millipore), anti–Tra‐1‐60 (Millipore), and anti–Tra‐1‐81 (Millipore). In immunofluorescence staining for cardiac markers, monoclonal anti–α‐actinin (Sigma), monoclonal anti‐cTnT (Thermo Scientific), polyclonal anti‐cTnT (Santa Cruz), monoclonal anti–myosin light chain (MLC)2a (Synaptic Systems), polyclonal anti‐MLC2v (ProteinTech Group), anti‐ANP (Santa Cruz), anti–cMyBP‐C (Santa Cruz), polyclonal anti–cMyBP‐C motif (supplied by C. Witt University of Heidelberg, Heidelberg, Germany), and anti–nuclear factor of activated T cells (NFAT)c4 (Santa Cruz) were used. The isotype‐specific secondary antibodies, Alexa Fluor 488 chicken anti‐rabbit IgG, Alexa Fluor 594 goat anti‐mouse IgG1, Alexa Fluor 488 goat anti‐rat IgM, Alexa Fluor 594 goat anti‐mouse IgM, Alexa Fluor 488 goat anti‐mouse IgG, Alexa Fluor 594 goat anti‐mouse IgG2b, Alexa Fluor 594 chicken anti‐goat IgG, and Alexa Fluor 555 goat anti‐rabbit IgG, were all obtained from Invitrogen. The tested drugs included endothelin‐1 (1.0, 10, 100, or 1000 nmol/L), angiotensin II (100 nmol/L), insulin‐like growth factor 1 (100 nmol/L), phenylephrine (0.05 mmol/L), BQ‐123 (250 nmol/L), and BQ‐788 (100 nmol/L) (all from Sigma).
Tanaka A., Yuasa S., Mearini G., Egashira T., Seki T., Kodaira M., Kusumoto D., Kuroda Y., Okata S., Suzuki T., Inohara T., Arimura T., Makino S., Kimura K., Kimura A., Furukawa T., Carrier L., Node K, & Fukuda K. (2014). Endothelin‐1 Induces Myofibrillar Disarray and Contractile Vector Variability in Hypertrophic Cardiomyopathy–Induced Pluripotent Stem Cell–Derived Cardiomyocytes. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 3(6), e001263.
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10

Pharmacological Agents Preparation

Cited 2 times
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Histamine, compound 48/80, endothelin-1, chloroquine, and GSK1016790A (GSK101) were purchased from Sigma. GSK2193874 (GSK219) was obtained from Tocris, U0126 was from Selleckchem, and GSK205 was synthesized (26 (link), 33 (link)). All were dissolved in sterile normal saline except that GSK101 and GSK205 were dissolved in DMSO (20 mm in stock) and further diluted until use.
Chen Y., Fang Q., Wang Z., Zhang J.Y., MacLeod A.S., Hall R.P, & Liedtke W.B. (2016). Transient Receptor Potential Vanilloid 4 Ion Channel Functions as a Pruriceptor in Epidermal Keratinocytes to Evoke Histaminergic Itch. The Journal of Biological Chemistry, 291(19), 10252-10262.
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