The effects of repeated endotoxin exposure on placental structure in sheep.

Intrauterine infection has been associated with fetal brain injury and preterm birth. We have recently shown that repeated exposure to bacterial endotoxin leads to hypoxia and brain injury in the preterm ovine fetus and we considered it possible that endotoxin could also damage the placenta. Our aim therefore was to assess placental structure following repeated exposure to endotoxin. Endotoxin was administered on 3-5 occasions (1 microg/kg, i.v.) over 5 days from 95-99 days of gestation (term approximately 147 days) to 6 fetal sheep and placental structure assessed at 105 days. In LPS-exposed animals there was a 17 per cent reduction (P<0.05) in placental weight and the average cross-sectional area of placentomes was reduced (P<0.05) by 20 per cent. In addition, all LPS-exposed placentae showed significant injury as evidenced by calcium deposits associated with areas of infarcted tissue. We conclude that repeated endotoxin exposure results in damage to the placenta which could lead to persistent alterations in placental exchange function.

Prolonged reductions in placental blood flow and cerebral oxygen delivery in preterm fetal sheep exposed to endotoxin: possible factors in white matter injury after acute infection.

OBJECTIVE: Endotoxin causes hypoxemia and white matter injury in the preterm ovine fetus. Because cerebral hypoxia could contribute to brain injury, our objective was to determine the effects of endotoxin on regional cerebral oxygen (O(2)) delivery. To investigate causes of fetal hypoxemia, we also measured placental blood flow. METHODS: We administered endotoxin (lipopolysaccharide, LPS) at 1 microgram/kg (intravenously) to 11 catheterized fetal sheep at approximately 0.7 of term; controls (n = 7) received saline. We measured fetal cerebral blood flow (CBF) and placental blood flow using microspheres, arterial blood gases, arterial pressure, and heart rate. RESULTS: Seven fetuses survived LPS administration (LPS-S) and four died. LPS-S fetuses were hypoxemic at 4-8 hours after LPS. Fetal hemoglobin concentration and hematocrit increased by about 14% at 4 hours after LPS exposure, and mean arterial pressure decreased significantly from 4-8 hours. After LPS, CBF did not change significantly, but total cerebral O(2) delivery decreased by 35.7% at 4 hours and by 28.3% at 8 hours. O(2) delivery to cerebral white matter decreased below pre-LPS values at 4 hours (-35.9%) and 8 hours (-28.6%) after LPS. Relative to pre-LPS values, placental blood flow decreased by 53.3% at 4 hours and 43.0% at 8 hours after LPS. CONCLUSIONS: Immature fetal sheep exposed to LPS had profound reductions in placental blood flow and cerebral O(2) delivery, which could contribute to fetal brain injury. Reduced O(2) delivery to white matter was similar to that in other brain regions. Mechanisms that enable fetal CBF to increase in hypoxemic conditions were apparently ineffective in the presence of LPS.

Stimulation of lung growth in fetuses with lung hypoplasia leads to altered postnatal lung structure in sheep.

Increased lung expansion in the fetus stimulates lung growth and is being trialed clinically to reverse severe fetal lung hypoplasia. Our aim was to examine the effects of increased fetal lung expansion in the presence of lung hypoplasia on lung structure in sheep at term and 8 weeks after birth. Lung hypoplasia was induced in 15 fetal sheep by continuous drainage of tracheal fluid, commencing at approximately 113 days of gestation (term, approximately 148 days). In 10 of these fetuses, tracheal obstruction (TO) was performed from 137-147 days of gestation (treated lung hypoplasia, TLH), while lung liquid drainage continued until term in the remaining 5 fetuses (untreated lung hypoplasia, ULH). Lung tissues were obtained from 5 TLH, 5 ULH, and 5 control lambs at birth, and from 5 TLH and 5 control lambs at 8 weeks after birth. At birth, alveolar number, surface area, and interalveolar wall thickness were not different between TLH and control lambs, whereas airspace diameter was greater in TLH lambs (72.7 +/- 3.0 microm) than in controls (58.4 +/- 4.3 microm). Diameters of airspaces were not different between ULH and control lambs; however, alveolar numbers and surface area were reduced, while interalveolar wall thickness was increased in ULH lambs compared to controls. At 8 weeks after birth, alveolar number (928.0 +/- 66.1 x 10(6)) and surface area (30.3 +/- 2.2 m(2)) in TLH lambs were lower, whereas interalveolar wall thickness (83.0 +/- 3.1 microm) was greater than in control lambs (2,263.6 +/- 261.6 x 10(6), 46.7 +/- 4.8 m(2), and 68.6 +/- 2.1 microm, respectively). Our data show that TO restores most aspects of lung structure to normal in fetuses with lung hypoplasia but leads to altered alveolar development. The presence of fewer, larger alveoli in postnatal TLH animals may predispose these animals to respiratory complications during later life.

Effects of fetal growth restriction on lung development before and after birth: a morphometric analysis.

Our aim was to determine the effects of fetal growth restriction (FGR) during late gestation on the structure of the lungs in the fetus near term and at 8 weeks after birth. The studies were performed using two groups of pregnant sheep and their offspring. In both groups, FGR was induced by umbilico-placental embolisation (UPE); for fetal studies, UPE was performed from 120 days of gestation until 140 days (term, approximately 146 days), when fetuses were killed for tissue analysis. For postnatal studies, UPE continued from 120 days until delivery at term; postnatal lambs were killed at 8 weeks after birth for tissue analysis. UPE led to a thicker pulmonary blood-air barrier at 140 days of gestation and this difference, which was due to a thickened basement membrane, was still present at 8 weeks after birth. At 8 weeks, we also observed a smaller number of alveoli per respiratory unit, thicker interalveolar septa, and a greater volume density of lung tissue in FGR lambs compared to controls. These changes would be expected to impair gas exchange and alter the mechanical properties of the lungs. Our data show that structural alterations in the lungs induced by placental insufficiency were more evident at 8 weeks of postnatal age than near term, indicating that the effects of FGR on the lung may become more serious with age and may affect respiratory health later in life.

Assessment of learning ability and behaviour in low birthweight lambs following intrauterine growth restriction.

The present study used behavioural tasks to assess learning ability and behaviour in postnatal lambs, and to examine the effects of low birthweight (LBW) and age on subsequent performance. It was hypothesized that intrauterine growth restriction (IUGR) and LBW lead to learning and behavioural deficits in the early postnatal period. IUGR and LBW were induced by umbilico-placental embolization from 120 days of gestational age (g.a.) to the onset of labour. Behavioural studies were performed on 6 LBW and 6 control lambs between 2 and 6 weeks after birth. LBW lambs were born at 139+/-1 days g.a. (2.4+/-0.2 kg) and control lambs were born at 149+/-1 days g.a. (4.5+/-0.4 kg). Three tasks were used to assess the learning ability and behaviour of the lambs: a simple maze, an obstacle course, and a T-maze. LBW lambs took longer to complete the simple maze at all ages, and made a greater number of errors at Week 1 of testing compared to control lambs; the total trial duration and number of errors decreased with age for both groups. In the obstacle course, the times taken to complete the first and third trials were used for analysis; a decrease in trial time and the number of errors from Trial 1 to Trial 3 were indications of the lamb's ability to learn how to negotiate the objects within the course. LBW lambs recorded longer trial durations for the first trial at Week 5 of testing, and for the third trial at Week 4. LBW lambs made more errors for the first trial at Week 5 of testing than control lambs. In the T-maze, there was no significant effect of treatment or age. It was concluded that differences between the groups may have been the result of LBW lambs being prematurely born. The value of these tasks in the assessment of learning ability and behaviour in young lambs is discussed.

Postnatal development of arterial pressure: influence of the intrauterine environment.

A substantial number of epidemiological studies have shown that small size at birth is associated with an increased risk of developing hypertension and metabolic dysfunction later in life; however these associations have not been found in all studies. In animals, several models have been used to investigate the effects of perturbations to the fetal environment on later arterial pressure, with differing effects on size at birth and arterial pressure. Ovine models include maternal dietary manipulations, antenatal glucocorticoid exposure, and restriction of placental size and function. In our laboratory, we have induced late gestational placental insufficiency and growth restriction in sheep by umbilico-placental embolisation; during the early postnatal period the growth restricted lambs remained small and were hypotensive relative to controls. More recent long-term studies indicate that these growth restricted animals were able to catch up in body weight within the first postnatal year; however, their arterial pressure remained lower than that of controls throughout the first 2 postnatal years (deltaMAP, -4.2 +/- 1.4 mmHg). This relative hypotension may be due to altered vascular or cardiac development resulting from increased vascular resistance or nutrient restriction during fetal life. As late gestational placental insufficiency led to a persistent reduction in arterial pressure from birth to adulthood, our findings do not support the hypothesis that restricted fetal growth per se leads to hypertension after birth. It is likely that the effects of a prenatal compromise on postnatal arterial pressure will vary depending on the nature of the associated developmental perturbations and their gestational timing.

Differential effects of prenatal exposure to dexamethasone or cortisol on circulatory control mechanisms mediated by angiotensin II in the central nervous system of adult sheep.

Prenatal exposure to elevated maternal glucocorticoids (dexamethasone (DEX) or cortisol (CORT)) for 2 days early in pregnancy can 'programme' alterations in adult offspring of sheep, including elevated arterial pressure. DEX treatment also results in greater angiotensin II type 1 (AT1) receptor expression in the medulla oblongata in late gestation fetuses than in saline (SAL)- or CORT-exposed animals. We hypothesized that this would result in functional changes in brainstem angiotensinergic control of cardiovascular function in DEX- but not CORT-exposed animals. To test this hypothesis, cardiovascular responses to intracerebroventricular (I.C.V.) angiotensin II were examined in adult male offspring exposed to DEX (0.48 mg h(-1); n = 7), CORT (5 mg h(-1), n = 6) or SAL (n = 9) from 26 to 28 days of gestation. Increases in mean arterial pressure during i.c.v. infusion of angiotensin II (1 or 10 microg h(-1)) were significantly greater in the DEX group (10 +/- 1 mmHg at 1 microg h(-1)) compared with SAL (6 +/- 1 mmHg) or CORT (6 +/- 1 mmHg) animals (P < 0.05). I.C.V. infusion of the AT1 antagonist losartan significantly decreased cardiac output and heart rate in DEX animals, but not in SAL or CORT animals. Thus, increased expression of brainstem AT1 receptor mRNA after prenatal DEX is associated with increased responsiveness of cardiovascular control to activation of brain AT receptors by exogenous and endogenous angiotensin II. The altered role of the brain RAS in sheep exposed prenatally to DEX was not observed in sheep exposed prenatally to cortisol, suggesting these two glucocorticoids have distinct programming actions.

Renal and amniotic fluid responses to umbilicoplacental embolization for 20 days in fetal sheep.

We determined the effects of placental insufficiency induced by umbilicoplacental embolization on fetal renal function and amniotic fluid volume and composition. Pregnant ewes underwent surgery at 115 +/- 2 days after mating (term approximately 147 days) for implantation of fetal vascular, bladder, and amniotic sac catheters. We studied five fetuses from 120 to 140 days during umbilicoplacental embolization and six control fetuses. Umbilicoplacental embolization reduced fetal arterial partial pressure of oxygen from 24.1 +/- 0.5 mmHg (pretreatment) to 14.6 +/- 0.2 mmHg. Fetal body weights were reduced to 80% of control values. Urine production and glomerular filtration rate in treated fetuses were significantly lower than in controls at 135 days of gestation. Amniotic fluid volume was not different between embolized and control animals. Fetal urine production in treated fetuses, when adjusted for body weight, was not different from that in control fetuses. We conclude that, in fetal growth restriction, reduced kidney weight, rather than hypoxemia per se is responsible for reduced urine production, which, if severe and prolonged, may contribute to oligohydramnios.

Swallowing of lung liquid and amniotic fluid by the ovine fetus under normoxic and hypoxic conditions.

OBJECTIVE: The lungs of the mammalian fetus secrete large volumes of fluid daily. The purpose of this study was to estimate the fraction of the lung liquid that is swallowed as it exits the fetal trachea versus that which enters the amniotic fluid under normoxic and hypoxic conditions. STUDY DESIGN: In chronically catheterized fetal sheep at 119 to 133 days' gestation the volume of fluid swallowed by the fetus was monitored five times per day for three consecutive 24-hour periods: control, hypoxia, and recovery. The Na+, K+, and Cl- concentrations of the swallowed fluid, lung liquid, and amniotic fluid were measured simultaneously. The fraction of the swallowed fluid that originated from the lungs or amniotic fluid was calculated from 24-hour average compositions and the assumption that the fetus swallowed only amniotic fluid and lung liquid. RESULTS: During the control, hypoxia, and recovery periods the fetuses swallowed 264 +/- 43 (SE), 92 +/- 23, and 271 +/- 24 ml/kg of fetal weight per day, respectively. As determined from Cl- concentrations, this swallowed fluid was composed of 17.7% +/- 2.7%, 24.8% +/- 5.8%, and 11.9% +/- 3.4% lung liquid, respectively, with the remainder being amniotic fluid. Throughout the three 24-hour observation periods there was an inverse relationship between the net 24-hour swallowed volume and the fraction of the swallowed fluid that originated from the lungs. Calculations based on Na+ concentrations yielded essentially the same results with slightly more scatter, whereas calculations based on K+ concentrations were unreliable. CONCLUSIONS: (1) Chloride concentrations provide the best of the three index values for a compositional analysis of fluids swallowed by the fetus. (2) Under normoxic conditions around 18% of swallowed fluid is derived from the fetal lungs. (3) On the basis of published fluid secretion rates for the fetal lung, an average of 50% of the liquid that exits the fetal trachea is swallowed and the rest mixes with the amniotic fluid.

The effects of twenty-four hours of reduced uterine blood flow on fetal fluid balance in sheep.

OBJECTIVE: Our aim was to determine the effects of a sustained reduction in uteroplacental perfusion, leading to fetal hypoxia, on determinants of amniotic fluid volume in sheep. STUDY DESIGN: Surgery was performed on five pregnant ewes 110 to 116 days after mating. At 127.3 +/- 2.2 days uterine blood flow was reduced for 24 hours, which reduced fetal SaO2 from 61.9% +/- 1.2% to 24.9% +/- 0.8%. RESULTS: Fetal urine production was increased from a control value of 193.0 +/- 24.0 ml/kg per 24 hours to 279.3 +/- 30.0 ml/kg per 24 hours during periods of reduced uterine blood flow and remained above control values for up to 48 hours after the reduced uterine blood flow period. A substantial loss of fetal water and electrolytes occurred through urine, which was associated with changes in the composition of fetal plasma and fetal tracheal, fetal swallowed, and amniotic fluids. Fetal swallowing was reduced throughout the reduced uterine blood flow period from a control value of 200.8 +/- 56.0 ml/kg per 24 hours to 32.7 +/- 8.4 ml/kg per 24 hours and returned to control levels after the cessation of the reduced uterine blood flow. CONCLUSION: We conclude that 24 hours of reduced uterine blood flow causes major changes in fetal renal function and fetal swallowing that, in spite of an expected reduction in lung liquid production, would increase the flow of fluid and electrolytes from the fetus into the amniotic sac.

Endocrine responses of fetal sheep to prolonged hypoxemia with and without acidemia: relation to urine production.

Our aim was to examine the endocrine changes associated with alterations in fetal urine production during 24 h of hypoxemia induced by either reduced uterine blood flow (RUBF) or maternal N2 inhalation (N2). In contrast to RUBF, which caused a diuresis, N2 caused a transient antidiuresis; during both posthypoxemia periods (RUBF and N2), fetal urine production was increased. RUBF, but not N2, was associated with a transient acidemia. Fetal plasma arginine vasopressin (AVP) and atrial natriuretic factor (ANF) concentrations increased during RUBF and were inversely correlated to pH; there were no detectable AVP or ANF responses to N2. Fetal prostaglandin E2 (PGE2) increased during the hypoxemia and posthypoxemia periods induced by both methods, but RUBF caused the greater increase. AVP and PGE2 concentrations were positively correlated with urine production. Fetal arterial blood pressure increased during RUBF but not N2. During RUBF, the increases in AVP and PGE2 concentrations and/or fetal arterial blood pressure may have contributed to the diuresis. During N2, we suggest that low, but increased, levels of AVP may have caused the transient antidiuresis, whereas the diuresis observed during both posthypoxemia periods may have been mediated by elevated PGE2 concentrations and/or increased fetal arterial blood pressure.

Postnatal outcomes in term and preterm lambs following fetal growth restriction.

1. Epidemiological evidence indicates that low birthweight increases the risk of a number of adult-onset diseases. It is now apparent that many babies with a low birthweight may have been subjected to a combination of reduced growth rates in utero as well as preterm birth. However, the long-term effects of preterm birth following intra-uterine growth restriction (IUGR) are unknown. Thus, our objectives were: (i) to identify prenatal factors associated with preterm birth in IUGR fetuses; and (ii) to characterize postnatal effects of preterm birth following IUGR. 2. We studied pregnant sheep and their offspring, in which fetal growth was restricted by umbilico-placental embolization during late gestation. Some of these animals were born at term (146 +/- 1 days) and some were born prematurely (139 +/- 1 days). In both groups, we have conducted longitudinal studies of postnatal respiratory function, cardiovascular function and learning ability up to 6-8 weeks of age. 3. Before birth, IUGR fetuses born prematurely (P-IUGR) were more hypoxaemic and acidaemic and had higher haemoglobin concentrations than both control fetuses and IUGR fetuses born at term (T-IUGR). In P-IUGR fetuses, plasma cortisol concentrations increased earlier than in the two other groups. The P-IUGR lambs had lower birthweights than T-IUGR lambs and both groups of IUGR lambs remained lighter than controls for 8 weeks. 4. After birth, P-IUGR lambs were hypoxaemic compared with T-IUGR and control lambs. Pulmonary diffusing capacity (adjusted for lung volume) was significantly lower in both groups of IUGR lambs than in controls, with P-IUGR lambs having lower values than T-IUGR lambs. Lung compliance (adjusted for lung volume), was not different between P-IUGR and control lambs, but values were higher in T-IUGR lambs than in control and P-IUGR lambs. Chest wall compliance (adjusted for lung volume) was higher in both groups of IUGR lambs than in controls. 5. During the 8 week postnatal study period, both groups of IUGR lambs had lower mean arterial pressures than control lambs; this relative hypotension was greatest in P-IUGR lambs. 6. In tests of learning ability, P-IUGR lambs took longer to complete a simple maze task at all ages and, in the second postnatal week, made a greater number of errors compared with controls. In an obstacle course, P-IUGR lambs recorded longer trial durations; they also made more errors than control lambs. 7. We conclude that preterm birth in the presence of late- gestational placental insufficiency and IUGR can result in specific effects on respiratory and cardiovascular development after birth, in addition to the effects of IUGR alone.

Changes in alveolar epithelial cell proportions during fetal and postnatal development in sheep.

Basal lung expansion is an important determinant of alveolar epithelial cell (AEC) phenotype in the fetus. Because basal lung expansion increases toward term and is reduced after birth, we hypothesized that these changes would be associated with altered proportions of AECs. AEC proportions were calculated with electron microscopy in fetal and postnatal sheep. Type I AECs increased from 4.8 +/- 1.3% at 91 days to 63.0 +/- 3.6% at 111 days of gestation, remained at this level until term, and decreased to 44.8 +/- 1.8% after birth. Type II AECs increased from 4.3 +/- 1.5% at 111 days to 29.6 +/- 4.1% at 128 days of gestation, remained at this level until term, and then increased to 52.9 +/- 1.5% after birth. Surfactant protein (SP)-A, -B and -C mRNA levels increased with increasing gestational age before birth, but the changes in SP expression after birth were inconsistent. Thus before birth type I AECs predominate, whereas after birth type II AECs predominate, possibly due to the reduction in basal lung expansion associated with the entry of air into the lungs.

Effects of intrauterine growth restriction on lung liquid dynamics and lung development in fetal sheep.

OBJECTIVE: The aim of this study was to determine the effects of intrauterine growth restriction on fetal lung liquid and lung development. STUDY DESIGN: Intrauterine growth restriction was induced in 7 fetal sheep from 120 to 140 days' gestation (term, approximately 147 days' gestation) by umbilicoplacental embolization. We used 6 control fetuses. Volumes and production rates of fetal lung liquid were measured, and lungs were removed post mortem (140 days' gestation) for analysis of concentrations of deoxyribonucleic acid, protein, and messenger ribonucleic acid for surfactant proteins A, B, and C. RESULTS: Umbilicoplacental embolization induced fetal hypoxemia, hypoglycemia, and intrauterine growth restriction. At 140 days' gestation lung weight was reduced by 34%, and the fetal lung liquid production rate (15.9 +/- 1.8 mL/h for intrauterine growth restriction vs 24.8 +/- 3.9 mL/h for control) and volume (110.9 +/- 16.3 mL for intrauterine growth restriction vs 178.1 +/- 11.9 mL for control) were reduced in the intrauterine growth restriction group. After adjustment for body weight, however, values were not different from those in the control group. Pulmonary deoxyribonucleic acid and plasma cortisol concentrations were elevated by intrauterine growth restriction, but levels of messenger ribonucleic acid for surfactant proteins were unchanged. CONCLUSION: In intrauterine growth restriction, lung liquid and lung growth were proportionate to body weight, and surfactant protein expression was unaffected. Alterations in lung deoxyribonucleic acid concentrations suggest that the lungs may be structurally immature.

Effects of chronic placental insufficiency on brain development in fetal sheep.

Clinical evidence has linked intrauterine compromise such as fetal hypoxemia to poor neurologic outcome in the newborn. In this study we examined the effects of inducing chronic fetal hypoxemia by impairment of placental function on brain development in fetal sheep. Placental insufficiency was induced from 120 to 140 d of gestation (term = 145-148 d) by injection of microspheres into the umbilical circulation in five fetal sheep. Fetal partial pressure of oxygen, PaO2, was reduced from 24.1 +/- 0.5 mm Hg before embolization to 14.8 +/- 0.4 mm Hg after embolization (p < 0.05). In another three fetuses a similar level of hypoxemia (PaO2, 13.8 +/- 0.4 mm Hg) occurred spontaneously. At 140 d of gestation the fetal brains were perfused with fixatives and compared with five control fetuses for the assessment of structural and immunohistochemical alterations. Hypoxemic fetuses demonstrated severe gliosis in the cerebral cortex and reduced myelination of subcortical white matter as visualized by glial fibrillary acidic protein and myelin basic protein staining, respectively (p < 0.05). White matter lesions were observed in two fetuses. The diameter of cerebral capillaries was increased in hypoxemic fetuses (p < 0.05), but there was no change in the number of nitric oxide synthase immunoreactive cells. Growth of neuronal processes was affected in the cerebellum, where there was also a reduction in the number of Purkinje neurons (p < 0.05). These results show that a prolonged period of placental insufficiency, resulting in moderate fetal hypoxemia during the last third of gestation, can affect neurodevelopmental processes that occur late in gestation such as myelination and growth of the cerebellum. This prenatal damage could affect neural connectivity and have functional consequences after birth.

Effects of prolonged hypoxemia on fetal renal function and amniotic fluid volume in sheep.

OBJECTIVE: Our purpose was to determine the effects of prolonged hypoxemia on fetal renal function and amniotic fluid volume and composition. STUDY DESIGN: Twelve pregnant ewes underwent surgery at 115 +/- 2 days after mating (term approximately 147 days) for the implantation of fetal vascular, bladder, and amniotic sac catheters. At 125 +/- 1 days seven fetuses were studied during 6 days of hypoxemia and five control fetuses were studied over six days of normoxemia. Index values of fetal renal function and amniotic fluid volume were measured. RESULTS: During hypoxemia fetal SaO2 and PaO2 were reduced from 60.9% +/- 1.6% and 21.9 +/- 0.6 mm Hg to 29.6% +/- 3.8% and 14.9 +/- 0.8 mm Hg, respectively. Fetal hypoxemia was associated with a transient acidemia (arterial pH 7.29 +/- 0.02) at 4 hours. There were no sustained alterations in fetal urine production (9.5 +/- 0.8 ml/hr/kg) or glomerular filtration rate (1.3 +/- 0.1 ml/min/kg) during hypoxemia. In control fetuses the amniotic fluid volume increased over 7 days, from 717 +/- 169 ml to 1031 +/- 147 ml, whereas in the hypoxemic fetuses it did not change (741 +/- 68 ml) over the same period. CONCLUSION: During prolonged fetal hypoxemia in the absence of acidemia, fetal urine production is maintained, whereas the normal gestational increase in amniotic fluid volume is prevented, raising the possibility that intramembranous reabsorption of amniotic fluid is increased by hypoxemia.

Alterations in fetal urine production during prolonged hypoxaemia induced by reduced uterine blood flow in sheep: mechanisms.

1. Our aim was to identify mechanisms whereby prolonged fetal hypoxaemia alters renal function and urine production in fetal sheep. 2. Fetal hypoxaemia was induced for 24 h by reducing uterine blood flow at 129.0 +/- 2.1 days of gestation (term 145-147 days), causing a reduction in fetal arterial O2 saturation (SaO2) from 52.5 +/- 2.3 to 22.0 +/- 1.3% (P < 0.05). This hypoxaemia was initially associated with a mild acidaemia (pH 7.23 +/- 0.03). 3. The glomerular filtration rate (GFR) increased from a control value of 1.8 +/- 0.3 mL/min per kg to a maximal value of 2.8 +/- 0.6 mL/min per kg (P < 0.05) at 4-5 h of hypoxaemia, returning to control levels by 6-9 h of hypoxaemia. After 4 h of hypoxaemia renal blood flow was no different to control values (144 +/- 8 mL/min per 100 g kidney weight) but after 24 h of hypoxaemia it had increased to 190 +/- 8 mL/min per 100 g kidney weight (P < 0.05). Fractional reabsorption of Na+ in the proximal tubules decreased from a control value of 81.5 +/- 2.2 to 65.2 +/- 3.9% at 2-3 h of hypoxaemia (P < 0.05) and remained reduced (68.5 +/- 3.1%) at the end of hypoxaemia (P < 0.05). Fetal mean arterial pressure transiently increased (P < 0.05) but returned to control values by 4-5 h of hypoxaemia. Fetal renal vascular resistance was not significantly altered during hypoxaemia. Fetal urine production increased from a control value of 12.3 +/- 2.1 mL/h per kg to a maximal value of 19.1 +/- 4.2 mL/h per kg at 4-5 h of hypoxaemia (P < 0.05) and returned to control by 24 h of hypoxaemia. 4. Our results indicated that prolonged fetal hypoxaemia leads to the inhibition of Na+ reabsorption in the proximal portion of the renal tubules. Changes in GFR induced by hypoxaemia were similar to those in fetal urine production and were not associated with changes in renal blood flow. We conclude that prolonged fetal hypoxaemia affects renal haemodynamics and the reabsorptive capacity of the renal tubules, resulting in a diuresis.

Relation between damage to the placenta and the fetal brain after late-gestation placental embolization and fetal growth restriction in sheep.

OBJECTIVE: Our aim was to determine the effects of 30 days of placental insufficiency on fetal brain development and to relate placental damage to the degree of fetal brain injury. STUDY DESIGN: Umbilicoplacental embolization was induced from 110 to 140 days of gestation (term, 147 days) in 7 fetal sheep, such that fetal arterial oxygen saturation was maintained at 50% of pre-umbilicoplacental embolization values. Six control fetuses were used. At 140 days the fetal brains and placentas were subjected to structural and histochemical analysis. RESULTS: During umbilicoplacental embolization, fetal arterial oxygen saturation, PaO(2), and pH were reduced (P <.05). Thirty days of umbilicoplacental embolization caused a decrease in cross-sectional area of the placentome (P <.05), with 20% of tissue showing damage. All umbilicoplacental embolization fetuses were growth restricted and had brain damage, most prominently in the cerebral white matter. There was no relation between the extent of placental damage and the severity of fetal brain damage. CONCLUSIONS: The absence of a correlation between damage to the placenta and fetal brain is likely to be caused by variations between individuals in (1) the amount of placenta that is required to be functionally damaged to achieve the prescribed level of hypoxemia and (2) the response of the fetal brain to that level of hypoxemia.

Placental insufficiency and fetal growth restriction lead to postnatal hypotension and altered postnatal growth in sheep.

Low birth weight has been associated with elevated arterial pressure in later life but mechanisms are unknown. Our aim was to determine the effects of low birth weight resulting from intrauterine growth restriction (IUGR) on fetal and postnatal arterial pressures and the potential roles of circulating cortisol and renin. We induced IUGR by umbilico-placental embolization (UPE) in fetal sheep from 120 d of gestation until birth (approximately 147 d); postnatal lambs (8 IUGR, 8 controls) were studied for 8 wk. Fetal and postnatal arterial pressures were measured and blood samples taken for measurement of gas tensions, cortisol concentrations and renin activity. In IUGR fetuses, mean arterial pressure (MAP) initially increased with UPE, but near term was not different to values in controls. IUGR lambs weighed 33% less than controls at birth and remained lighter than controls during the 8 postnatal weeks; their growth pattern was different to that of controls. IUGR lambs had lower MAP than controls, and this relative hypotension (-4 mm Hg) persisted throughout the 8 postnatal weeks. Covariate analysis showed that the relative hypotension of IUGR lambs could have resulted from their smaller size. Plasma cortisol concentrations were not different between IUGR and control animals before or after birth. Plasma renin activity was not different in postnatal IUGR lambs compared with controls. Thus, postnatal cortisol and renin levels were not consistent with the development of hypotension or hypertension. We conclude that late gestational IUGR in sheep leads to relative hypotension in the early postnatal period, probably a result of reduced body size.

Compromised respiratory function in postnatal lambs after placental insufficiency and intrauterine growth restriction.

Epidemiologic studies have shown persistent effects of low birth weight on respiratory function and lung health, but underlying mechanisms are not understood. Our aim was to determine the effects of intrauterine growth restriction (IUGR), a major cause of low birth weight, on postnatal respiratory function. IUGR was induced by umbilico-placental embolization during late gestation in chronically catheterized sheep. Umbilico-placental embolization was performed between 120 d of gestation and term ( approximately 146 d) during which fetuses were hypoxemic and hypoglycemic relative to controls. Umbilico-placental embolization led to a 48% reduction in birth weight compared with controls, and throughout the postnatal study period IUGR lambs (n = 8) remained lighter than controls (n = 8). Respiratory function was repeatedly studied in lambs for 8 wk after birth; during this period, IUGR lambs were mildly hypoxemic and tended to be hypercapnic compared with controls. In IUGR lambs, relative to controls, O(2) consumption (mL/min/kg) and minute ventilation (mL/kg) were increased and pulmonary diffusing capacity (adjusted for functional residual capacity) was decreased. Functional residual capacity, measured by helium dilution, and total lung capacity (measured at 30 cm H(2)O) were smaller in IUGR lambs than in controls. When adjusted for functional residual capacity, static lung compliance was reduced and chest wall compliance was increased in IUGR lambs. At 8 wk, pulmonary DNA and protein concentrations were decreased in IUGR lambs relative to controls. We conclude that restriction of fetal growth by placental insufficiency induces alterations in the lungs and chest wall that result in persistent impairments in respiratory function during early postnatal life.