Superimposition of length fluctuations on contracted ASM have shown to reduce active force and stiffness. This effect is usually attributed to disruption of the actomyosin crossbridge cycle; however no direct experimental data is available to support this hypothesis. This in vitro study investigated the effect of the mechanical strains on 1) the ASM reactivity and 2) on the actin-myosin crossbridges. Experiments were carried out on maximally contracted bovine ASM subjected to length strains at various frequency in the range from 10 to 100Hz, superimposed on normal tidal stretches (frequency 0.33Hz, amplitude 4%). An organ bath system was used to apply strains and measure the force; immunofluorescence technique was performed to assess the crossbridges. The results show that superimposed length strains increase breathing relaxation effect with an optimal effect obtained at 50Hz. The cholinergic stimulation promotes actin-myosin connection, and length stretches promote the detachment of those crossbridges.
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