Mice were euthanized by CO2 narcosis, and the thoracic aorta was cut into 2-mm aortic rings, mounted in organ baths of a Mulvany-Halpern myograph at a resting tension of 13 kPa in Krebs with or without l -arginine (100 μmol/L; Sigma, UK) and contracted with of endothelin (ET)-1 (30 pmol/L to 30 nmol/L; Sigma, UK) or the thromboxane mimetic U46619 (1 nmol/L to 1 μmol/L). After washing, tissues were contracted with an EC80 concentration of U46619 (30–100 nmol/L) followed by acetylcholine (1 nmol/L to 10 μmol/L) or sodium nitroprusside (SNP; 1 pmol/L to 10 μmol/L). For ET-1 studies, vessels were preincubated with the NOS inhibitor NG-nitro-l -arginine methyl ester (100 μmol/L; Sigma, UK) to exclude any effect of changes in the NO/ADMA pathway. Force was recorded via a PowerLab/800 (AD Instruments Ltd., UK) and analyzed by using Chart 6.0 acquisition system (AD Instruments Ltd, UK).
Chart 6.0 acquisition system
Manufactured by ADInstruments
Sourced in United Kingdom
The Chart 6.0 acquisition system is a data acquisition software solution developed by ADInstruments. It is designed to capture, display, and analyze experimental data from various types of laboratory equipment. The core function of the Chart 6.0 acquisition system is to provide a platform for users to record, monitor, and manipulate data collected during experiments.
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2 protocols using chart 6.0 acquisition system
1
Aortic Ring Contractility Assay
2
Vascular Function Optimization Protocol
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C57 Black 4 mice (6–10 weeks) were killed by CO2 narcosis and aorta removed, cleaned of connective tissue and cut into 1.5 mm rings before being mounted in Mulvany-Halpern myograph organ baths containing a physiological salt solution (PSS), as we have described previously30 (link). In order to optimize the stability of vascular function over the 8-h time course, diclofenac (1 µM) and cycloheximide (1 µM) were added to the PSS to block vasoactive prostanoids and induction of vasoactive genes (e.g., NO synthase) respectively. Vessels were contracted with an EC80 concentration of U46619 (10 nM). Once a stable baseline was obtained, sildenafil (10 µM), nanoMIL-89 (10 µg/ml), or Sil@nanoMIL-89 (10 µg/ml) was added to the PSS, and vascular tone monitored for 8 h. Responses in vessels incubated in PSS served as controls. Force was recorded via a PowerLab/800 (AD Instruments Ltd., UK) and analysed using Chart 6.0 acquisition system (AD Instruments Ltd., UK). All studies using animals or animal tissues/organs were conducted in accordance with UK Home Office Animals (Scientific Procedures) Act 1986.
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