3D visualisation and quantification by microcomputed tomography of late gestational changes in the arterial and venous feto-placental vasculature of the mouse.

This study evaluates microcomputed tomography (micro-CT) as a method to obtain quantitative three-dimensional (3D) information on the arterial and venous vasculature of the mouse placenta. Surface renderings at embryonic days (E) 13.5, 15.5, and 18.5 (full term) revealed that the arterial and venous vasculature branched within the chorionic plate whereas only the arterial vasculature deeply penetrated the placenta. Umbilical vessel diameters measured by micro-CT did not significantly differ from those measured non-invasively in vivo by ultrasound biomicroscopy. Variability in umbilical diameters, and surface area and volume measurements of arterial and venous vascular trees due to experimental error was low relative to biological variability, and significant inter-litter differences within gestational ages were detected. Furthermore, umbilical vessel diameter increased significantly and incrementally to an arterial diameter of 0.631+/-0.009 mm and a venous diameter of 0.690+/-0.018 mm at E18.5. Umbilical vein diameter was 3-9% greater than the artery, and both were significantly correlated with embryonic body weight (R> or =0.96). Surface area and volume were determined for vessels greater than the minimum resolvable diameter of 0.03 mm which therefore excluded capillaries. Arterial surface area and volume were unchanged from E13.5-15.5 but then more than doubled at E18.5 (to 170+/-13 mm(2) and 7.2+/-0.8mm(3), respectively). Venous surface areas and volumes changed similarly with development although surface areas were lower than their arterial counterparts. We conclude that micro-CT has sufficient accuracy and precision to quantify late gestational changes in the 3D structure of the arterial and venous vasculature of the mouse placenta.

Role of the L-arginine nitric oxide pathway in hypoxic fetoplacental vasoconstriction.

The role of nitric oxide in hypoxic fetoplacental vasoconstriction (HFPV) was investigated using dually perfused human placental cotyledons. Standard medium (Earle's salt solution with added dextran and L-arginine) was equilibrated with 95 per cent O2 and 5 per cent CO2 (maternal side) and 94 per cent N2 and 6 per cent CO2 (fetal side). Part 1 consisted of perfusion for 1 h, then maternal perfusate equilibrated with a 95 per cent N2 and 5 per cent CO2 for 20 min (hypoxia), and then the original perfusion conditions resumed for 40 min. In part 2, this sequence was repeated with standard medium alone (n = 6), or with added N-nitro-L-arginine methyl ester (L-NAME) (n = 6), or L-NAME and nitroglycerin (n = 6). When standard medium was used throughout, basal fetal perfusion pressure (30 +/- 2 mmHg) and the hypoxia-induced increase in perfusion pressure (18 +/- 1 mmHg) did not change significantly between parts 1 and 2. L-NAME increased basal perfusion pressure from 33 +/- 3 to 56 +/- 2 mmHg whereas perfusion pressure remained unchanged with L-NAME and nitroglycerin or nitroglycerin alone. The hypoxic vasoconstriction observed during part 1 in the L-NAME (14 +/- 3 mmHg) and the L-NAME with nitroglycerin groups (18 +/- 2 mmHg) was abolished during part 2 (to - 4 +/- 1 and 0.4 +/- 0.5 mmHg, respectively) whereas nitroglycerin alone significantly blunted the response (21 +/- 3 to 6 +/- 1 mmHg). Results suggest that a reduction in basal NO release mediates hypoxic fetoplacental vasoconstriction in the perfused human placental cotyledon in vitro.

Low dose indomethacin via fetal vein or cerebral ventricle stimulates breathing movements in fetal sheep.

Indomethacin, a prostaglandin synthesis inhibitor, transforms normally intermittent fetal breathing into nearly continuous breathing. If indomethacin acts in brain, we hypothesized that indomethacin would stimulate breathing at a lower dose when given via the lateral cerebral ventricle (i.c.v) than via the fetal jugular vein (i.v.). Chronically catheterized fetal sheep were studied at 0.88 of gestation (128 days). A 2-h control period was followed by an indomethacin infusion at 7 micrograms.kg-1 fetal body weight over the first 10 min of each hour for 3 h then at 28 micrograms.kg-1 over the first 10 min of each hour for the next 3 h, either i.c.v. (n = 7) or i.v. (n = 6). Plasma prostaglandin E2 concentrations decreased similarly by both routes, although the decrease was only significant for the i.c.v. route. Indomethacin at 7 micrograms.kg-1.h-1 did not change fetal breathing activity by either route within 3 h. During the 2nd and 3rd h at 28 micrograms.kg-1.h-1, breathing incidence increased (from approximately 35 to approximately 80% time), breath amplitude and rate increased, and arterial O2 content decreased slightly (all changes significant). ANOVA revealed no significant differences in responses evoked by the two routes. Fetal arterial pressure, heart rate, Pco2, and pH were unchanged (both doses, both routes). The indomethacin dose (105 micrograms.kg-1 over 6 h) was > or = 10-fold lower than that used in previous studies on breathing in fetal sheep but similar to that used clinically to treat patent ductus arteriosus. We conclude that site(s) mediating effects of indomethacin on breathing respond to a low dose and are equally accessible by i.c.v. and i.v. routes.

Endothelin-1 constricts fetoplacental microcirculation and decreases fetal O2 consumption in sheep.

Endothelin-1 produced by umbilicoplacental tissues may regulate fetal placental perfusion. To investigate its site of action, we measured segmental resistance in this bed in unanesthetized fetal sheep near term during fetal endothelin-1 infusion. A 15-min intravenous infusion of endothelin-1 at 1 micrograms/min significantly increased fetal blood pressure in the aorta (+33%), cotyledon artery and vein, and inferior vena cava, and endothelin-1 decreased fetal heart rate (-40%). Vascular resistance in the placental microcirculation increased significantly (+332%), but smaller increases in resistance of the umbilical artery and vein were not significant. Nevertheless, the stiffness of the umbilical arterial wall appeared to increase because vascular input impedance increased significantly both at the heart rate frequency (+85%) and when averaged > 2 Hz (characteristic impedance; +138%). Mean blood flow in the umbilical artery decreased by 64%, and the flow pulsatility index increased 137% (P < 0.05 for both). Despite the large decrease in placental perfusion, there was no significant change in descending aortic oxygen tension or oxygen content, because fetal oxygen consumption was reduced by 40%. We conclude that endothelin-1 is a potent constrictor of the placental microcirculation in sheep. Endothelin-1 also decreases fetal oxygen consumption by an unknown mechanism.

Pressure wave propagation and input impedance in thoracic aorta of conscious newborn sheep.

Aortic hemodynamics were examined in eight conscious newborn sheep. Flow and pressure in the thoracic aorta and pressure in the distal abdominal aorta were measured under control conditions and during pressure changes caused by bottle feeding or during intravenous infusions of nitroprusside, norepinephrine, or angiotensin II. Vasoconstriction affected aortic impedance, pressure wave amplification, and wave velocity similarly whether induced by feeding or by drugs. Central hemodynamics in the lamb were surprisingly similar to hemodynamics in the sheep fetus despite major changes in cardiovascular function at birth, largely because pressure-related increases in pulse wave velocity postpartum compensated for increased arterial lengths and increased heart rate. Wave reflection effects on pressure-flow relations were more prominent during vasoconstriction and less prominent during nitroprusside. Wave reflections in both lambs and fetal sheep return to the heart in early diastole; therefore, they do not add to ventricular afterload. Early diastolic return of reflected waves characterizes adults of many species, and demonstration of the phenomenon throughout the perinatal period reinforces arguments for its adaptive value.

Pulsatile pressure-flow relations and pulse-wave propagation in the umbilical circulation of fetal sheep.

The relations between pulsatile pressures and flows in the umbilico-placental circulation have been investigated using chronically instrumented fetal sheep. Under resting conditions, mean arterial pressure fell by 30 +/- 6%, from 44 +/- 2 to 31 +/- 2 mm Hg between the aortic termination and the arteries feeding the cotyledons, and pressure waves were substantially damped during propagation between the two recording sites. This high flow resistance and wave attenuation are attributed to the very thick walls and extreme length of the umbilical arteries. Unique relations between pulsatile components of pressure and flow, characterized as vascular impedance spectra, were also observed. At rest, impedance to pulsatile flow was only slightly below resistance to steady flow, and impedance phase was positive at low frequencies. Pulse-wave reflections had more modest effects in this bed than others. Thus, oscillations in impedance spectra and percent wave transmission with increasing frequency, which are widely accepted manifestations of wave reflections, were relatively small. Positive impedance phases at low frequencies indicated that novel mechanisms influence phase relations between pressure and flow. A significant vascular compliance residing in the peripheral vascular beds could account for this findings. The vasodilator nitroprusside enhanced wave-reflection effects, whereas the vasoconstrictor angiotensin II reduced these effects. These changes were opposite to the effects of vasoactive substances in other systems, probably because these drugs act predominantly on the supply (umbilical) arteries rather than on the peripheral placental vasculature. When peripheral vascular resistance was selectively elevated by infusing 50-microns microspheres, reflection effects were enhanced: the pressure pulse in the umbilical artery was transmitted without attenuation, or was amplified, and impedance spectra more closely resembled patterns typical of other vascular beds. Specifically, impedance modulus fell sharply with increasing frequency, and impedance phase was negative at low frequency. In addition, we observed coordinated oscillations in impedance modulus and phase that are characteristic of beds that exhibit wave-reflection effects. These findings indicate that the specialized anatomy and control mechanisms observed in the umbilical circulation result in unique hemodynamic function, in which wave-propagation effects exert influences not readily predictable from studies on other systems.

An electron microscope study of fibril: matrix arrangements in high- and low-crimp wool fibres.

The crimped structure of wool fibres is generally associated with a bilateral arrangement of ortho- and paracortical cells. The most obvious difference between these cell types is in the arrangement and relative proportions of microfibril and matrix proteins that constitute the fibre cortex. In the low-crimp fibres examined there is a poorer expression of bilateral cortical asymmetry compared with the high-crimp wools together with a higher proportion of intermediate-staining mesocortical cells. These mesocortical cells exhibit much more regular arrays of microfibrils than paracortical cells. It is suggested that the packing arrangement of microfibrils in all three cell types is basically hexagonal and the variation observed in mature cells is a function of the fibril:matrix ratio.

Influence of nutrition on the crimping rate of wool and the type and proportion of constituent proteins.

When the nutritional level of sheep is restricted, the staple crimp frequency of the resultant fleece increases substantially whereas the cystine and high-sulphur protein contents decrease. This is in marked contrast to the direct relationship between crimp frequency and cystine content among sheep. These observations can be reconciled by assuming that variations in crimp frequency are attributable solely to a combination of follicle shape and fibre length growth rate without recourse to the more generally accepted theories relating to the proportion and distribution of ortho- and paracortical cells in the firbre cortex. The major portion of the decrease in the cystine content of high-crimp wools is due to the decreased synthesis of a specific protein fraction (ultra-high-sulphur protein) as would be expected from the results of dietary supplementation experiments. Low-crimp wools do not appear to contain this protein fraction and in this respect they may differ from high-crimp wools.