Co-ordination of electrical activity in the myometrium of pregnant ewes.

The co-ordination of spontaneous and evoked electromyographic (EMG) activity was examined in the uterus of ewes from Day 100 of pregnancy and during labour. During pregnancy the onset of bursts of EMG activity was synchronous to within 2-3 min when recorded by electrodes at any recording site. Individual EMG spikes, whether occurring spontaneously or evoked by electrical stimulation of the myometrium, appeared to propagate between recording electrodes that were 1-3 cm apart, in the longitudinal or circular orientation of the uterus, but not when the electrodes were separated by more than 3 cm. Apparent conduction velocity in the longitudinal orientation was 7 cm sec-1 during pregnancy and increased to 13 cm sec-1 during labour. No consistent pattern could be distinguished in the sites at which EMG activity was first observed at any stage of pregnancy studied. The nature of propagation of EMG activity remains unresolved but it is clear that it is not initiated at a single site in the myometrium.

Influence of upper respiratory tract on liquid flow to and from fetal lungs.

The experiments were designed to determine the influence of the upper respiratory tract (URT) on liquid flow in the fetal trachea. This flow probably influences pulmonary distension, which is thought to be a major determinant of prenatal lung development. In six fetal sheep the URT could be bypassed by connecting the lower trachea, via an external flowmeter, to a cannula in the amniotic sac. In confirmation of our earlier findings, when the URT was in circuit, the mean rate of tracheal efflux was greater during episodes of fetal breathing movements (FBM) [mean 13.8 +/- 2.6 (SE) ml/h] than during apneic periods (mean 3.2 +/- 1.0 ml/h). When the URT was bypassed there was a reversal of net tracheal flow during FBM episodes (mean 19.6 +/- 5.6 ml/h toward the lungs); during apnea there was a much greater rate of efflux (mean 33.1 +/- 10.2 ml/h) than when the URT was in circuit. Nonlabor uterine contractions were associated with an increased rate of efflux during apnea only when the URT was bypassed. We conclude that during fetal life the URT imposes an essentially unidirectional flow of pulmonary liquid away from the lungs, preventing ingress of amniotic fluid and maintaining constancy of composition of liquid in the developing airways. By retarding outward flow during periods of apnea and thoracic compression and by preventing net influx during episodes of FBM, the URT has the probable effect of maintaining the volume and composition of liquid in the fetal airways within narrow limits.

Changes in the innervation and catecholamine concentrations in the myometrium of pregnant and non-pregnant sheep.

The innervation of the myometrium in virgin, pregnant and post-partum ewes was examined using the SPG histofluorescence method and a high-performance liquid chromatographic catecholamine assay. In non-pregnant ewes, fluorescent axons were visible in all regions of the uterus. At 50 days of pregnancy the innervation was unchanged, but at 100 days axons were scarce over the whole uterus with the exception of the tubal extremities of the uterine horns. Noradrenaline concentrations were also significantly lower in late pregnancy, and significant variations occurred in different regions of the uterus, with the greatest concentrations present in the tubal extremities. At 13-16 weeks post partum, the density of innervation was variable, although the noradrenaline concentration was only slightly less than in virgin animals. Dopamine was also present in substantial quantities, but the mean concentrations remained unchanged during pregnancy.

Upper airway resistances in fetal sheep: the influence of breathing activity.

Fetal breathing movements (FBM) and lung liquid volume are known to affect lung development, but little is known about mechanisms controlling movement of liquid through the upper respiratory tract (URT). Therefore we measured resistances of the URT in 8 unanesthetized fetal sheep during late gestation while FBM were monitored from pressures in the lower trachea or from electromyogram of respiratory muscles. URT resistance to liquid flow toward the amniotic sac increased from 3.5 +/- 1.9 Torr X ml-1 X min during episodes of FBM to 21.1 +/- 5.7 Torr X ml-1 X min during apnea. Laryngeal resistance during apnea was greater (P less than 0.001) than supralaryngeal resistance in each of six fetuses in which URT resistance was partitioned. Fetal paralysis abolished the increase in laryngeal resistance to efflux that was previously related to the high-voltage electrocortical state and apnea. We were unable to quantify URT resistance to fluid movement toward the lungs because the larynx acted as a valve, permitting flow toward the lungs only in the presence of FBM. The supralaryngeal portion of the URT also apparently acts as a valve, normally preventing the entry of amniotic fluid into the pharynx. These findings help to explain our earlier observations that efflux of liquid from the fetal lungs is greater during episodes of FBM than during apnea.

The regulation of flow of pulmonary fluid in fetal sheep.

The flow of fluid within the trachea of fetal sheep (121-135 days) has been measured using a newly developed flowmeter. The flowmeter, which is basically a miniature servo-controlled peristaltic pump, is connected in series with an extra-corporeal tracheal loop. Integrated tracheal flow was measured for periods of at least 8 h in 5 control fetuses during which we determined its relationship to fetal breathing movements detected by EMG's of inspiratory muscles or tracheal pressure fluctuations. The overall flow of fluid away from the lungs was 14.4 ml/h; on average outward flow was 5.3 times greater during episodes of breathing movements than during apnea. Interruption of the motor innervation of the larynx in 5 fetuses led to a 25% reduction in mean overall flow and, compared with control fetuses, there was a reduction in net flow associated with breathing movements and an increased flow during apnea. These findings suggest that tracheal flow is normally retarded by a laryngeal mechanism during apnea, giving rise to an elevated pressure within the trachea and probably resulting in increased pulmonary distension. Paralysis of the fetus with gallamine triethiodide reduced the flow of liquid from the lungs and abolished variations in flow rate related to fetal electrocortical states.

Ingestion in fetal sheep and its relation to sleep states and breathing movements.

Swallowing was recorded electromyographically in unanaesthetized fetal sheep in utero during the last third of gestation. In control fetuses bouts of swallowing were, on average, 2.0 min in duration and occurred at intervals of 2.31 h. Swallowing bouts were associated with a low-voltage electrocorticogram, eye movements and, in 82% of instances, with phasic activity in the dorsal neck muscles. These data indicate that fetuses are not in quiet (non-rapid eye movement; n.r.e.m.) sleep when swallowing and also suggest that they are not in r.e.m. sleep as conventionally defined; we consider that fetuses may be in a state of heightened activity resembling wakefulness. Swallowing bouts were always accompanied by fetal breathing movements. When a swallow coincided with an inspiratory effort the diaphragm was briefly inhibited. Evidence was obtained that bouts of fetal ingestive activity were not dependent upon outpourings of fluid from the lungs and were not significantly influenced by volume information from the gastric compartments.

Composition and volume of fluid swallowed by fetal sheep.

Oesophageal cannulations were performed in seven fetal sheep between 109 and 128 d of gestation. Fluid swallowed by the fetuses was collected externally and periodically returned to the fetal stomach by a peristaltic pump. The volume of fluid swallowed daily was calculated from the movement of water in a parallel pump circuit. Samples of oesophageal fluid were removed from the collection bag at the same time as tracheal and amniotic fluid samples. Oesophageal fluid was very viscous and was usually intermediate in colour between tracheal and amniotic fluids. After reduction of viscosity by dithiothreitol, the compositions of the three fluids were analysed. Mean concentrations of Na+, Cl- and protein, and mean osmolarity of oesophageal fluid, were intermediate between values obtained in tracheal and amniotic fluids. [K+] was often greater in oesophageal fluid than would be expected by the relative contribution of tracheal and amniotic fluids as indicated by [Na+] and [Cl-] in the three fluids. It is suggested that the high viscosity of oesophageal fluid and its relatively high [K+] may result from fetal salivary secretions. Ionic concentrations in individual sets of samples show that the proportions of tracheal and amniotic fluids swallowed by the fetus are variable from day to day. The mean daily volumes of fluid swallowed by the seven fetuses ranged from 98 to 577 ml, and are in agreement with previously published data.

Development of myometrial electrical activity during the first half of pregnancy in the sheep.

Uterine myoelectrical activity was recorded in seven pregnant sheep covering the period between 13 and 75 days post-coitum. Activity in the myometrium was present at day 13 and took the form of intermittent spikes of low amplitude. Bursts of spikes of irregular duration became noticeable between days 25 and 40 but most were not coordinated throughout the myometrium. Coordinated bursts of myoelectrical activity, which could be recorded at several sites simultaneously, first appeared between 40 and 65 days. These bursts had similar characteristics to the myoelectrical activity associated with uterine contractions during the last third of gestation. The myoelectrical activity showed a progressive increase in amplitude during the first half of gestation. There was no relationship between plasma progesterone levels and the increase in amplitude or appearance of coordinated bursts of uterine activity.

Relationship between electrical activity of the uterus and surgically isolated myometrium in the pregnant and nonpregnant ewe.

Rhythmic contractions occur in the sheep uterus at oestrus and during pregnancy from about 65 days to term (145 days). To define factors responsible for these contractions we have examined and quantified the degree of synchronization of electrical activity in the uterus and isolated segments of myometrium in 3 types of experiments. In the first, a segment of myometrium was totally separated from the uterus. After a period of 9-16 days the isolated tissue developed a typical pattern of uterine activity which showed no significant degree of synchronization with EMG bursts in the body of the uterus. During labour, the isolated tissue showed changes in activity similar to those observed in the uterus. In the 2nd experiment, the tubal end of one of the uterine horns was severed from the uterus, but a connection was retained with the uterus via the oviduct and ovarian blood vessels. Activity in the partly isolated segment remained in synchrony with the uterus. In a 3rd experiment, impulse propagation through nerves and smooth muscle to the tip of a horn was disrupted by severing 'the tip' from the uterus while its blood supply from the ovarian vessels was retained. The blood vessels were momentarily frozen, and denervation confirmed by monoamine histofluorescence. In 5 out of 6 animals the operated tissue showed activity that was not synchronous with the rest of the uterus. These data indicate that: (1) isolated uterine muscle in vivo has rhythmicity resembling that of intact myometrium and (2) systemic or local circulating factors are not responsible for synchronizing uterine activity before parturition, although circulating factors do play a major role in increasing the uterine activity which occurs at parturition and at oestrus, and (3) hydraulic continuity between different regions of the uterus is not essential for maintaining co-ordinated activity.