Temporal and spatial variations in spontaneous Ca events and mechanical activity in pregnant rat myometrium

Abstract

Objectives The aim of this study was to investigate the temporal and spatial characteristics of spontaneous Ca signals in pregnant rat myometrium. Study design Confocal imaging of longitudinal strips of 21-day pregnant rats loaded with the Ca sensitive indicator Fluo-4, was combined with measurements of mechanical activity in uterine smooth muscle cells, in situ and freshly isolated. Results Our results show that the Ca transients in pregnant uterine tissue are composed of Ca spikes, which are associated with the spike-like action potentials. There is large variation in the pattern of spontaneous activity in myometrium, ranging from non-propagating Ca spikes confined to individual smooth muscle cells, through to regional and global propagating Ca spikes. Irrespective of the pattern of activity displayed, the Ca signals were always in the form of Ca spikes, singularly or in bursts. These Ca spikes did not show fixed initiations sites, propagated in longitudinal and transverse directions from the initiation regions, and had a variable pattern of propagation in preparations which were not synchronously active. In preparations which showed synchronous activity, Ca spikes singularly or bursts propagated mainly in the transverse direction from the initiation regions. The amplitude of force generated by single spikes was dependent on the number of bundles recruited by the propagating Ca spike within the strip, and was about 30–40% of the maximal force produced by carbachol or high-K stimulation. If Ca spikes appeared in the form of bursts they generated longer lasting fused contractions, the amplitudes of which were dependent on the number and the frequency of Ca spikes in the burst. Conclusions Longitudinal myometrium from pregnant rats generates spontaneous Ca spikes which vary in their initiation sites, spatial spread and frequency and are associated with the spike-like action potentials. They are sensitive to the L-type Ca channel blocker, nifedipine. Contractile activity was dependent on the spatial spread of individual Ca spikes and when fully synchronized, produced single submaximal phasic contraction. The number and frequency of bursts of Ca spikes controlled the amplitude and duration of contraction.