Wire myograph

Manufactured by Danish Myo Technology

Sourced in Denmark, United States

The Wire Myograph is a lab equipment used for the study of blood vessel reactivity. It measures the contractile and relaxation properties of small arterial and venous tissue samples. The device uses wire supports to mount and monitor the changes in the diameter of the vessel segment under various experimental conditions.

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42 protocols using wire myograph

Endothelium-Dependent Relaxation in Mesenteric Arteries

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Male 5-month-old Wistar Kyoto rats were killed with an overdose of a mixture of the anesthetics ketamine (140 mg/kg intramuscularly) and xylazine (40 mg/kg intramuscularly). Isolated third branches of mesenteric arteries were mounted on a wire myograph (Danish Myo Technology, Aarhus, Denmark) to measure isometric tension. Arteries were superfused with warm (37°C) physiological saline solution of the following composition (mM): 119 NaCl, 4.7 KCl, 1.18 KH₂PO₄, 1.17 MgSO₄, 25 NaHCO₃, 5.5 glucose, and 1.6 CaCl₂, pH 7.4, adjusted with NaOH. After the normalization procedure, arteries were left to equilibrate for 1 hour at 37°C before subsequent evaluation. Prior to each experiment the integrity of the endothelium was confirmed by contracting arteries with 4 μm phenylephrine followed by 1 μm endothelium-dependent vasodilator acetylcholine (Ach). Vessels that failed to produce at least 80% relaxations were eliminated.
To evaluate the effect of SiNPs on endothelium-dependent relaxation, arteries were contracted with 4 μm phenylephrine and then relaxed with Ach in a dose-response manner (0.1 nm–10 μm) in the absence of and after incubation with SiNPs (2 μg/mL, 10 μg/mL, and 50 μg/mL).

Nemmar A., Albarwani S., Beegam S., Yuvaraju P., Yasin J., Attoub S, & Ali B.H. (2014). Amorphous silica nanoparticles impair vascular homeostasis and induce systemic inflammation. International Journal of Nanomedicine, 9, 2779-2789.

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Vascular Reactivity Assay: Mesenteric and Aortic Rings

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Mesenteric and aortic ring vascular reactivity was studied on a wire myograph (Danish Myo Technology, Arhus, Denmark) as previously described [33 (link)]. Arteries were bathed in a physiological salt solution (PSS: 119 mM NaCl, 4.7 mM KCl, 14.9 mM NaHCO₃, 1.2 mM MgSO₄.7H₂O, 2.5 mM CaCl₂, 1.18 mM KH₂PO₄, and 5.5 mM glucose under a pH 7.4, PO₂ 160 mm Hg, PCO₂ 37 mmHg) continuously bubbled with 95% O₂ and 5% CO₂. After an equilibration period of at least 20 min. under the optimal passive tension, two successive contractions in response to the combination of KCl (100 mM) depolarization and serotonin (5-HT; 10 μM) (Sigma-Aldrich) were used to test the maximal contractile capacity of the vessels. After washing, the endothelial function was assessed by testing the relaxing effect of acetylcholine (Ach 1 μM) after precontraction by 1 μM 5-HT. Following a 20-min. washout period, concentration-response curves to 5-HT were elicited by a cumulative administration of the vasoconstrictor agonist (1 nM to 100 μM) to vessels with their endothelium submitted or not to a specific iNOS inhibitor, N-([3-(Aminomethyl)phenyl]methyl) ethanimidamide dihydrochloride (1400W, 100 μM; Sigma-Aldrich), or a specific COX-2 inhibitor, N-(2-cyclohexyloxy-4-itrophenyl)methanesulfonamide (NS-398, 10 μM; Sigma-Aldrich). The inhibitor was added in the bath 30 min. before the addition of 5-HT.

Boisramé-Helms J., Meyer G., Degirmenci S.E., Burban M., Schini-Kerth V., Cynober L., De Bandt J.P., Hasselmann M, & Meziani F. (2016). “Immunonutrition” Has Failed to Improve Peritonitis-Induced Septic Shock in Rodents. PLoS ONE, 11(1), e0147644.

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Vascular Function Assessment in Murine Femoral Arteries

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After mice were euthanized by CO₂ inhalation, femoral arteries were removed and placed in oxygenated ice-cold Krebs solution that contained (mmol/L) 119 NaCl, 4.7 KCl, 2.5 CaCl₂, 1 MgCl₂, 25 NaHCO₃, 1.2 KH₂PO₄, and 11 D-glucose. Changes in isometric tone of the femoral arteries were recorded in wire myograph (Danish Myo Technology, Aarhus, Denmark). The vascular segments were stretched to an optimal baseline tension of 0.8-1 mN and then allowed to equilibrate for 1 h before the experiment commenced. Segments were first contracted with 60 mmol/L KCl and rinsed in Krebs solution. After several washouts, phenylephrine (10 μmol/L) was used to produce a steady contraction, acetylcholine (10 nmol/L to 30 μmol/L) was added cumulatively to induce endothelium-dependent vasodilatation on different segments. Endothelium-independent vasodilatation to SNP was performed in the presence of nitric oxide synthase inhibitor L-NAME (0.1 mmol/L), indomethacin (1 μmol/L), and 20 mmol/L KCl. Statistical significance was calculated either using the area under curve for each segment or indicated on the individual data points.

Wu Y., Tang X., Lee S., Hong H., Cao X., Lau C.W., Liu B., Chawla A., Ma R.C., Huang Y., Lui K.O, & Tian X.Y. (2022). Endothelial PPARδ facilitates the post-ischemic vascular repair through interaction with HIF1α. Theranostics, 12(4), 1855-1869.

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Vascular Reactivity Assessment in Mouse Aorta and Mesenteric Arteries

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The C57BL/6 mice were sacrificed by CO₂ anesthesia, and the mouse aortas or mesenteric resistance arteries were dissected out and placed in oxygenated ice-cold Krebs solution (NaCl: 119 mM, KCl: 4.7 mM, CaCl₂: 2.5 mM, MgCl₂: 1 mM, NaHCO₃: 25 mM, KH₂PO₄: 1.2 mM, and D-glucose: 11 mM). Ring segments of the thoracic aortas or mesenteric resistance arteries (~2 mm in length) were suspended between two tungsten wires (40 µm in diameter) in organ chambers (Multi Myograph System) filled with 5 mL oxygenated Krebs solution at 37 ℃. The changes in the isometric tone of the rings were recorded by wire myograph (Danish Myo Technology, Aarhus, Denmark). The rings were stretched to an optimal baseline tension of 3 mN (thoracic aortas) and 2 mN (mesenteric resistance arteries) and kept for equilibration for 1 h. The rings were contracted with a 60 mM KCl solution and rinsed in Krebs solution. Endothelium-dependent relaxations induced by acetylcholine (Ach, 10 nM to 10 µM, Sigma-Aldrich, St. Louis, MO, USA) or insulin (0.1 µM to 5 µM, Sigma-Aldrich, St. Louis, MO, USA) were determined in phenylephrine (Phe, 3 µM; Sigma-Aldrich, St. Louis, MO, USA) -precontracted rings. The relaxation was presented as a percentage reduction of phenylephrine-induced contraction [39 (link),40 (link)].

Kang L., Yi J., Lau C.W., He L., Chen Q., Xu S., Li J., Xia Y., Zhang Y., Huang Y, & Wang L. (2023). AMPK-Dependent YAP Inhibition Mediates the Protective Effect of Metformin against Obesity-Associated Endothelial Dysfunction and Inflammation. Antioxidants, 12(9), 1681.

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Assessing Vascular Reactivity ex vivo

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Saphenous arteries, muscular resistance arteries with a diameter of ∼250 µm that are well suited to assess vasomotor responses ex vivo[22] (link), [26] (link), were used to study endothelium-dependent vasodilatation. Animals were euthanized by CO₂/O₂ inhalation. The arteries were dissected free from surrounding fat and connective tissue, and were mounted in a wire myograph (Danish MyoTechnology, Aarhus, DK). Arterial segments (2 mm long) were distended to the diameter at which maximal contractile responses to 10 µM NA (noradrenaline) were obtained [22] (link), [27] (link). Optimal diameters (D_opt) and maximal contractile responses to NA for male mice are summarized in Table S2.

Chennupati R., Meens M.J., Marion V., Janssen B.J., Lamers W.H., De Mey J.G, & Köhler S.E. (2014). Endothelial Arginine Resynthesis Contributes to the Maintenance of Vasomotor Function in Male Diabetic Mice. PLoS ONE, 9(7), e102264.

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Vasoreactivity Assessment in Mouse Arteries

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Vasoreactivity was measured in wire myograph as described previously (12 (link), 13 (link)). The aorta or femoral arteries of Cdh5-Cre;Agrn^fl/fl mice were removed and dissected in oxygenated ice-cold Krebs solution that contained (in mmol/L): 119 NaCl, 4.7 KCl, 2.5 CaCl₂, 1 MgCl₂, 25 NaHCO₃, 1.2 KH₂PO₄, and 11 _D-glucose. Measurements of isometric tension were recorded in wire myograph (Danish Myo Technology). The arterial segments of aorta and femoral artery were respectively stretched to optimal baseline tension at 3 and 1 mN. After that, they were washed in Krebs solution three times and allowed to equilibrate for 1 h before being contracted with 60 mmol/L KCl to test viability. Endothelium-dependent relaxation (EDR) was measured by testing concentration responses to the cumulative addition of acetylcholine (ACh) in phenylephrine (Phe, 3 μmol/L) pre-contracted segments. Some arteries were incubated with NO synthase inhibitor L-NAME (100 μmol/L, 30 min) before testing vasodilation. Vasoconstriction induced by cumulative concentration of Phe was tested before and after L-NAME, which reflects basal NO production.

Ye P., Fu Z., Chung J.Y., Cao X., Ko H., Tian X.Y., Tang P.M, & Lui K.O. (2022). Endothelial Agrin Is Dispensable for Normal and Tumor Angiogenesis. Frontiers in Cardiovascular Medicine, 8, 810477.

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Vascular Reactivity Measurement Protocol

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Segments of the thoracic aorta and of the second‐order mesenteric arteries were mounted in a wire myograph (Danish Myo Technology, Aarhus, Denmark) as previously described.20 Briefly, 2 tungsten (for the mesenteric artery) or steel (for the aorta) wires were inserted into 2‐mm‐long arterial segments; 1 was fixed to a force transducer and 1 to a micrometer. The arterial segments were bathed in a physiological salt solution of the following composition: 130 mmol/L NaCl; 15 mmol/L NaHCO₃; 3.7 mmol/L KCl; 1.2 mmol/L KH₂PO₄; 1.2 mmol/L MgSO₄; 11 mmol/L glucose; 1.6 mmol/L CaCl₂; 5 mmol/L HEPES, pH 7.4; 160 mm Hg po₂; 37 mm Hg pco₂. Wall tension was applied as described previously.21 Arterial segment viability was tested using a potassium‐rich solution (KCl, 80 mmol/L). Endothelial function was then tested using acetylcholine (10⁻⁶ mol/L) after precontraction with phenylephrine (10⁻⁶ mol/L). After washout, a cumulative concentration‐response curve to acetylcholine (10⁻⁹ to 10⁻⁵ mol/L) and then to sodium nitroprusside (10⁻⁹ to 10⁻⁵ mol/L) was performed.

Guivarc'h E., Favre J., Guihot A., Vessières E., Grimaud L., Proux C., Rivron J., Barbelivien A., Fassot C., Briet M., Lenfant F., Fontaine C., Loufrani L., Arnal J, & Henrion D. (2020). Nuclear Activation Function 2 Estrogen Receptor α Attenuates Arterial and Renal Alterations Due to Aging and Hypertension in Female Mice. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 9(5), e013895.

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Contractile Function of Mouse Pulmonary Arteries

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Mice (25–30 g) were sacrificed by cervical dislocation, and small intrapulmonary arteries (IPA) of third order (100–150 μm in diameter) were isolated from the lung, dissected free of connective tissue, and placed in Krebs’ bicarbonate buffer solution (pH 7.4) containing the following (in millimolars): 118 NaCl, 4.7 KCl, 1.5 CaCl₂ × 2H₂O, 25 NaHCO₃, 1.1 MgSO₄, 1.2 KH₂PO₄, 5.6 glucose, and 10 HEPES. Then, the vessels were mounted on a wire myograph (Danish Myo Technology A/S, Aarhus, Denmark) and bathed in Krebs’ buffer solution at 37°C and an optimal passive tension of 3 mN. After 30 minutes of incubation, the arterial viability and equilibration were assessed by the stimulation of the vessels with repeated 10-minute exposures to KCl (60 mM; 60 K). For registration of vascular ring contractile activity and its following analysis, Chart 5.5.4 and LabChart Reader 8.1.9 (ADInstruments, Inc.) software were used. Vascular tension is presented as a percentage of the maximum steady-state contraction level obtained to the exposure to 60 K.

Kitagawa A., Jacob C., Jordan A., Waddell I., McMurtry I.F, & Gupte S.A. (2021). Inhibition of Glucose-6-Phosphate Dehydrogenase Activity Attenuates Right Ventricle Pressure and Hypertrophy Elicited by VEGFR Inhibitor + Hypoxia. The Journal of Pharmacology and Experimental Therapeutics, 377(2), 284-292.

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Isometric Tension Recording of Renal Arteries

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Segments of renal arteries (≈2 mm) were mounted in a wire myograph (Danish Myo Technology, Aarhus, Denmark) for isometric tension recording.^{28 (link)} Concentration-effect curves for methoxamine (30 nmol/L–30 µmol/L) or KCl (10–120 mmol/L) were constructed to evaluate arterial contractility after the experimental treatments. Data were recorded and analyzed using LabChart 7 (ADInstruments, Dunedin, New Zealand).

Barrese V., Stott J.B., Baldwin S.N., Mondejar-Parreño G, & Greenwood I.A. (2020). SMIT (Sodium-Myo-Inositol Transporter) 1 Regulates Arterial Contractility Through the Modulation of Vascular Kv7 Channels. Arteriosclerosis, Thrombosis, and Vascular Biology, 40(10), 2468-2480.

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Vascular Reactivity to Insulin and IGF-1

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Two-millimetre segments of first-order mesenteric arteries were harvested from LT HF and LF mice and mounted in a wire myograph (Danish Myo Technology A/S, Aarhus, Denmark) containing physiological buffer (mM): KCl 7.4, NaCl 118, NaHCO₃ 15, KH₂PO₄ 1.2, MgSO₄ 1.2, glucose 11, CaCl₂ 2.5, EDTA 0.023 at 37°C, 5% CO₂ and 95% O₂. Vessels were equilibrated at a resting lumen diameter of 0.9 × L100 (L100 represents vessel diameter under passive transmural pressure of 100 mmHg) in buffer for 30 min. Three potassium-induced constrictions were performed using high potassium buffer and vessels constricting less than 1 mN were excluded from the study. Vessels were pre-constricted with phenylephrine, at a dose yielding approximately 40% constriction obtained with high potassium buffer, and left to stabilise for 10 min. Relaxation to cumulative addition of either insulin (0.001 nM/1 pM to 1 µM) or IGF-1 (0.001 nM to 10 nM) was assessed in pre-constricted vessels. A time-matched control recording was also performed following the same protocol, without the addition of insulin or IGF-1. The contractile force of a vessel segment was recorded using PowerLab 4/25–LabChart7 acquisition system (ADInstruments, Oxford, UK).

Mughal R.S., Bridge K., Buza I., Slaaby R., Worm J., Klitgaard-Povlsen G., Hvid H., Schiødt M., Cubbon R., Yuldasheva N., Skromna A., Makava N., Skytte-Olsen G, & Kearney M.T. (2018). Effects of obesity on insulin: insulin-like growth factor 1 hybrid receptor expression and Akt phosphorylation in conduit and resistance arteries. Diabetes & Vascular Disease Research, 16(2), 160-170.

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