Fort 10g

The porcine coronary arterial rings were mounted using a wireless triangular support (Radnoti, Monrovia, CA, USA) in conjunction with a rigid L-shaped support. The triangular support was attached to a force-displacement transducer (FORT 10g, World Precision Instruments Inc., Sarasota, FL, USA) connected to an amplifier and a computer recording system (BIOPAC systems, Santa Barbara, CA, USA). Next, the coronary arterial rings were suspended in an organ bath containing 5 mL Krebs–Henseleit buffer and gassed with 95% O₂•5% CO₂. The final pH at 37°C was 7.40 ± 0.05.
The coronary arterial ring preparations were equilibrated at a resting tension of 2.0 g. After initial equilibration, the preparations were exposed to 60 mM KCl for 4 min. The experiments commenced after a 45-min equilibration period. For the relaxation measurements in the contracted arterial rings, the DRLE was added in a non-cumulative manner to U46619 (100 nM)-contracted muscle strips 15 min after the addition of U46619. The relaxation responses were represented as a percentage of U46619-stimulated contraction. Only one single dose response was studied with each preparation.

The mice were anesthetized with urethane (1.3 g/kg, S.C.) and their anesthesia state was confirmed through a gentle toe pinch. Subsequently, a laparotomy was meticulously performed to expose the bladder, following the same procedure used for viral vector injection. Then, a polyethylene catheter (KN-392, Natsume Seisakusho Co. Ltd, Tokyo, Japan) connected to a pressure sensor (BLPR2, World Precision Instruments, LLC, Sarasota, FL, USA) and a syringe pump (PHD 2000, Harvard Apparatus, Holliston, MA, USA) was inserted into the dome region of the bladder. After 2 h of recovery time post-catheterization, PBS or PBS diluted with 0.1% Acetic acid was infused into the bladder at a rate of 3 mL/h. To measure the voiding volume of the mice, a force transducer (FORT10G, World Precision Instruments, LLC, Sarasota, FL, USA) with a plastic cup was employed. Real-time data collection of intravesical pressure of the bladder and the voiding volume was conducted at a sampling rate of 500 Hz using a data acquisition system (Digidata 1440a, Molecular Devices, San Jose, CA, USA).

An electronic force transducer (FORT 10 g, World Precision Instruments) excited with ± 5 V produced by a BK Precision DC power supply was used to measure swelling of a single kernel held in a semi-micro acrylic spectrophotometer cuvette (Fig. 1a). A thin plastic cap on top of the kernel formed a flat surface to push a plastic rod with a conical tip up against the measuring leaf of the transducer after the addition of distilled water caused the kernel to swell (Fig. 1). The unfiltered output of the transducer, proportional to applied force, was digitized at a frequency of 1 Hz for 24 h after addition of water by a computer equipped with an A/D converter and custom software written in the LABVIEW (National Instruments) programming language. The 86,400 point time series was smoothed by the Sovitzky-Golay method with a window of 500 points using Origin (Microcal) analysis software to produce the traces shown in Fig. 1b. The transducer output increased linearly with known test masses used to convert output voltage to force units.