Si mb4 tissue bath system

Bladders were pinned on a small Sylgard block, and muscle was dissected free of the mucosal tissue. BSM strips were then cut longitudinally (2–3 mm wide and 5–7 mm long). Full thickness stomach strips of 5 mm width were cut from the fundus region in the direction of the longitudinal muscle. Full thickness jejunum was cut into ~3 mm rings. Muscle strips were mounted in an SI-MB4 tissue bath system (World Precision Instruments, Sarasota, FL, USA). Force sensors were connected to a TBM 4 M transbridge (World Precision Instruments), and the signal amplified by PowerLab (AD Instruments, Colorado Springs, CO, USA) and monitored through Chart software (AD Instruments). BSM strips were gently prestretched to optimize contraction force, then pre-equilibrated for at least 1 h. All experiments were conducted at 37 °C in physiological saline solution (in mM: 120 NaCl, 5.9 KCl, 1.2 MgCl₂, 15.5 NaHCO₃, 1.2 NaH₂PO₄, 11.5 Glucose and 2.5 mM CaCl₂) with continuous bubbling of 95% O₂/5% CO₂. Contraction force was sampled at 2000/s using Chart software. BSM tissue was treated with agonists or antagonists, and/or subjected to electrical field stimulation (EFS).

Bladders from male wild type and P2X₄−/− mice were pinned on a small Sylgard block and bladder mucosa was dissected away carefully. BSM strips were then cut longitudinally (2–3 mm wide and 5–7 mm long) and mounted in an SI-MB4 tissue bath system (World Precision Instruments, FL, USA). Force sensors were connected to a TBM 4 M transbridge and the signal amplified by Powerlab and monitored through Chart software. Contraction force was monitored dynamically with a sampling rate of 2000/s. BSM strips were gently pre-stretched to get optimized force and equilibrated for at least 1 h before any experiments. All experiments were conducted at 37 °C in physiological saline solution (PSS in mM: Na, 136.9; K, 5.9; Ca, 2.5; Mg, 1.2; Cl, 133.6; HCO₃, 15.5; H₂PO₄, 1.2; glucose, 11.5; pH 7.4), with continuous bubbling of 95% O2 and 5% CO2.

Bladders were pinned on a small Sylgard block, and the muscle was dissected free of the mucosal tissue. BSM strips were then cut longitudinally (two mm wide and seven mm long). Muscle strips were mounted in an SI-MB4 tissue bath system (World Precision Instruments, FL, USA). Force sensors were connected to a TBM 4M transbridge (World Precision Instruments), and the signal was amplified by PowerLab (AD Instruments, CO, USA) and monitored through Chart software (AD Instruments). BSM strips were gently prestretched to optimize contraction force and then pre-equilibrated for at least 1 h. All experiments were conducted at 37 °C in physiological saline solution (in mM: 120 NaCl, 5.9 KCl, 1.2 MgCl₂, 15.5 NaHCO₃, 1.2 NaH₂PO₄, 11.5 glucose, and 2.5 mM CaCl₂) with continuous bubbling of 95% O₂/5% CO₂. Contraction force was sampled at 2000/s using Chart software. BSM tissue was treated with agonists or antagonists and/or subjected to electrical field stimulation (EFS). BSM strip EFS was carried out using a Grass S48 field stimulator (Grass Technologies, RI, USA) using previously described standard protocols (Rajandram et al., 2016 (link); Chen et al., 2020 (link)).