Regulation Of Amniotic Fluid Volume Research Articles

Vascular endothelial growth factor activation of intramembranous absorption: a critical pathway for amniotic fluid volume regulation.

The purpose of this review is to propose a critical role for vascular endothelial growth factor (VEGF) in mediating the transfer of amniotic fluid from the amniotic compartment through the fetal membranes and fetal surface of the placenta into fetal blood. Experimental findings in humans and animal models on the action of VEGF in mediating fluid transfer are reviewed and interpreted in order to postulate a proposed mechanism for VEGF regulation of amniotic fluid absorption through the fetal membranes and placenta. Recent scientific advances suggest that up-regulation of VEGF gene expression in the amnion and chorion is associated with increased transfer of amniotic fluid into fetal blood. The possible mechanisms of action for VEGF appear to involve regulation of intramembranous blood vessel proliferation and membrane transport via passive permeation as well as nonpassive transcytotic vesicular movement of fluid. Currently evolving concepts suggest that amniotic fluid volume is regulated through modulation of the rate of intramembranous absorption of amniotic fluid by both passive and nonpassive mechanisms. The permeability factor VEGF appears to be a critical regulator of amniotic fluid transport in the fetal membranes.

Mechanisms for regulation of intramembranous amniotic fluid (AF) absorption: gene expression of prolactin (PRL) receptor nephrin

FLUID (AF) ABSORPTION: GENE EXPRESSION OF PROLACTIN (PRL) RECEPTOR NEPHRIN FATANEH AMIDI, DAVE GAYLE, SAM WANG, MICHAEL ROSS, Harbor-UCLA Medical Center, Department of Obstetrics and Gynecology, Torrance, CA OBJECTIVE: Although absorption of AF across the chorioamnion (intramembranous flow) is central to AF volume regulation, little is known regarding its regulation. Nephrin-like proteins regulate transcellular protein permeability in several organs at epithelial podocytes. PRL, having watermineral hormone properties, also has a potential role in AF volume. As amnion cells have similar structures as podocytes, we postulated that Neph1 and PRL may regulate fetal chorioamnion permeability and thus influence AF volume. We sought to determine the expression of Neph1 and PRL receptor genes in rat fetal membranes and placenta. Results are compared to beta-actinmRNA levels. STUDY DESIGN: Pregnant Sprague Dawley rats (n = 3) at gestation day 18 were anesthetized and the uterus exposed. Individual placentas and fetal chorioamnion were harvested and homogenized to isolate total RNA for mRNA determinations using real time RT-PCR. RESULTS: Fetal membranes expressed high levels of neph1 mRNA which was significantly greater than placental levels ((1.3 ± 0.3 vs 0.01 ± 0.01 AU, p = 0.03). Fetal membranes also had elevated levels of PRL receptor mRNA that were higher than placenta levels (0.21 ± 0.05 vs 0.05 ± 0.03 AU, p = 0.08). CONCLUSION: These finding indicate the expression of Neph1 and PRL in fetal membranes. We postulate that Neph1 and PRL regulate fetal membrane permeability to protein and electrolytes, respectively. Alterations in expression of these genes may lead to AF volume abnormalities.

Brain natriuretic peptide and endothelin-1 in the pathogenesis of polyhydramnios-oligohydramnios in monochorionic twins

Objective: We investigated the association between amniotic fluid levels of human brain natriuretic peptide, endothelin-1, and abnormal amniotic fluid volume in monochorionic twins with and without chronic twin-twin transfusion syndrome. Study Design: Amniotic fluid and fetal blood samples were obtained in utero or at cesarean delivery from monochorionic twins with (n = 20) or without chronic twin-twin transfusion syndrome (n = 10). Concentrations of atrial natriuretic peptide, human brain natriuretic peptide, and endothelin-1 (in picograms per milliliters) were determined by radioimmunoassay. Results: The amniotic fluid concentrations of human brain natriuretic peptide (P <.001) and endothelin-1 (P <.001) in the recipient fetuses were higher than the donor twins but were similar in the twins with no twin-twin transfusion syndrome. In the donor twins, amniotic fluid concentrations of human brain natriuretic peptide (P <.001) and endothelin-1 (P <.001) were lower than the twin pairs with no twin-twin transfusion syndrome. In both chronic twin-twin transfusion syndrome fetuses (P <.01) and fetuses with no twin-twin transfusion syndrome (P <.001), the amniotic fluid concentrations of human brain natriuretic peptide were high, although the concentrations of the endothelin-1 were lower than the fetal plasma concentrations. A positive association was present between amniotic fluid levels of human brain natriuretic peptide and endothelin-1 (R2 = 0.51, P <.001, n = 60). Amniotic fluid human brain natriuretic peptide (r = 0.67, P <.001) and endothelin-1 (r = 0.57, P <.01) levels of the recipient twins correlated with the amniotic fluid index. Conclusion: These data suggest that amniotic fluid concentrations of human brain natriuretic peptide and endothelin-1 were highest in the twins with polyhydramnios and lowest in the twins with oligohydramnios, which suggests the importance of these hormones in the regulation of amniotic fluid volume. (Am J Obstet Gynecol 2003;189:189-94.)

Regulation of amniotic fluid volume by intramembranous absorption in sheep:: Role of passive permeability and vascular endothelial growth factor

Objective: During long-term intravascular fluid infusion in the ovine fetus, a large increase in fetal urinary flow rate occurs while amniotic fluid volume increases only slightly because of increased intramembranous absorption. The current study tested the hypotheses that passive intramembranous permeability increases in response to fetal intravascular saline solution infusion and that the increased intramembranous absorption occurs in parallel with an increase in vascular endothelial growth factor gene expression in the amnion, chorion, and placenta. Study Design: Chronically catheterized fetal sheep that average 126 ± 1 (SE) days of gestation either were infused intravascularly with 7 L of normal saline solution over 3 days (n = 8 sheep) or served as time controls (n = 6 sheep). Amniotic fluid volume and fetal urinary flow rate were measured daily. Intramembranous diffusional permeability was estimated daily as being equal to the clearance of intra-amniotically injected technetium 99m. Vascular endothelial growth factor messenger RNA abundance in the amnion, chorion, and placenta was determined by Northern blot analysis. Statistical analyses included analysis of variance. Results: In the infused fetuses, amniotic fluid volume and urinary flow increased (P <.01) by 891 ± 144 mL and 3488 ± 487 mL per day, respectively, on infusion day 3 compared with no changes over time in the control fetuses. In the infused fetuses, estimated intramembranous absorption increased by 4276 ± 499 mL during the 3-day infusion. Intramembranous technetium 99m permeability was similar over time in the two groups. In the infused group, vascular endothelial growth factor messenger RNA levels in the amnion, chorion, and placenta increased 2- to 4-fold compared with the control group (P <.001). Conclusion: The up-regulation of vascular endothelial growth gene expression may mediate the increase in the intramembranous absorption that is induced by volume-loading diuresis; however, this does not occur by passive mechanisms. We speculate that vascular endothelial growth mediates the increased intramembranous absorption by increasing vesicular transport. (Am J Obstet Gynecol 2003;188:786-93.)

Neuronal NO modulates spontaneous and ANG II-stimulated fetal swallowing behavior in the near-term ovine fetus.

Spontaneous fetal swallowing occurs at a markedly higher rate compared with spontaneous adult drinking activity. This high rate of fetal swallowing is critical for amniotic fluid volume regulation. Central NO is critical for maintaining the normal rate of fetal swallowing, as nonselective inhibition of NO (with central N(G)-nitro-L-arginine methyl ester) suppresses spontaneous and angiotensin II (ANG II)-stimulated swallowing. We sought to differentiate the contributions of central endothelial vs. neuronal NO in the regulation of spontaneous and stimulated fetal swallowing, using a selective neuronal NO synthase (nNOS) inhibitor. Six time-dated pregnant ewes and fetuses were chronically prepared with fetal vascular and intracerebroventricular (i.c.v.) catheters and electrocorticogram (ECoG) and esophageal electromyogram electrodes and studied at 130 +/- 1 days of gestation. After an initial 2-h baseline period (0-2 h), the selective nNOS inhibitor N-propyl-L-arginine (NPLA) was injected i.c.v. (2-4 h). At 4 h, the dose of NPLA was repeated, together with ANG II, and fetal swallowing was monitored for a final 2 h. Four fetuses also received an identical control study (on an alternate day) in which NPLA was replaced with artificial cerebrospinal fluid (aCSF). Suppression of nNOS by i.c.v. NPLA significantly reduced mean (+/- SE) spontaneous fetal swallowing (1.35 +/- 0.12 to 0.50 +/- 0.07 swallows/min; P < 0.001). Injection of ANG II in the presence of NPLA had no dipsogenic effect on fetal swallowing (0.68 +/- 0.09 swallows/min). In the aCSF study, i.c.v. aCSF did not change fetal swallowing (0.93 +/- 0.10 vs. 0.95 +/- 0.09 swallows/min), whereas i.c.v. ANG II resulted in a significant increase in the rate of fetal swallowing (2.0 +/- 0.04 swallows/min; P = 0.001). We speculate that the suppressive dipsogenic effects of central NPLA indicate that spontaneous and ANG II- stimulated fetal swallowing is dependent on central nNOS activity.

Ovine fetal swallowing: expression of preterm neurobehavioral rhythms.

Fetal swallowing contributes importantly to amniotic fluid volume regulation and fetal gastrointestinal maturation. Near-term ovine fetal swallowing occurs in discrete bouts of activity (at approximately 30-min intervals) in association with fetal electrocortical voltage changes. Thus, swallowing rhythms have been hypothesized to be entrained to fetal neurobehavioral states. In the preterm ovine fetus, electrocortical activity does not demonstrate differentiation into high- and low-voltage periods until 120-130 days' gestation. We sought to quantify patterns of preterm (114 days, 0.75 gestation) ovine fetal swallowing activity and volume, and, in view of the lack of electrocortical pattern changes, to explore whether swallowing activity was regulated by an independent central pacemaker. Six singleton ovine pregnancies were chronically prepared with fetal and maternal femoral artery and vein catheters. Biparietal electrocortical electrodes were placed on the fetal skull. Following a minimum 5-day recovery period, fetuses were studied at 114 +/- 1 days. Patterns of fetal swallowing behavior were quantified by computer analysis of laryngeal-esophageal electromyography (EMG) and thoracic esophageal fluid flow during a 12-h period. Esophageal fluid flow was bidirectional, although antegrade flow predominated, leading to an average fluid acquisition rate of 13 +/- 3 ml/h (7.3 +/- 1.8 ml/h per kg) during the 12-h study (302 +/- 87 ml/day). Propagated esophageal EMG activity, representing coordinated 'swallows', averaged 56 +/- 6 swallows/h and correlated well with net esophageal fluid flow. 'Bouts' of swallowing activity (> or = 3 swallows/min) averaged 9 +/- 1 swallows/bout, lasted 1.8 +/- 1.4 min and accounted for 31 +/- 4% of the swallowed volume. Despite the absence of fetal electrocortical high-voltage/low-voltage transitions, there was a 26.1 +/- 3.9-min interval between periods of swallowing bout activity. Preterm (0.75 gestation) ovine fetal volume swallowed (302 ml/day) and volume swallowed for body weight (175 ml/day per kg) was significantly less than that previously noted at 0.85 gestation (831 ml/day, 274 ml/day per kg, respectively; p < 0.05) although the rates of swallowing activity were similar. The presence of swallowing bout activity at periodic intervals, in the absence of electrocortical differentiation, suggests an intrinsic central pacemaker regulating preterm fetal neurobehavior.

Ovine fetal swallowing: expression of preterm neurobehavioral rhythms

Objective: Fetal swallowing contributes importantly to amniotic fluid volume regulation and fetal gastrointestinal maturation. Near-term ovine fetal swallowing occurs in discrete bouts of activity (at approximately 30-min intervals) in association with fetal electrocortical voltage changes. Thus, swallowing rhythms have been hypothesized to be entrained to fetal neurobehavioral states. In the preterm ovine fetus, electrocortical activity does not demonstrate differentiation into high- and low-voltage periods until 120-130 days' gestation. We sought to quantify patterns of preterm (114 days, 0.75 gestation) ovine fetal swallowing activity and volume, and, in view of the lack of electrocortical pattern changes, to explore whether swallowing activity was regulated by an independent central pacemaker. Methods: Six singleton ovine pregnancies were chronically prepared with fetal and maternal femoral artery and vein catheters. Biparietal electrocortical electrodes were placed on the fetal skull. Following a minimum 5-day recovery period, fetuses were studied at 114 &#45 1 days. Patterns of fetal swallowing behavior were quantified by computer analysis of laryngeal-esophageal electromyography (EMG) and thoracic esophageal fluid flow during a 12-h period. Results: Esophageal fluid flow was bidirectional, although antegrade flow predominated, leading to an average fluid acquisition rate of 13 &#45 3 ml/h (7.3 &#45 1.8 ml/h per kg) during the 12-h study (302 &#45 87 ml/day). Propagated esophageal EMG activity, representing coordinated 'swallows', averaged 56 &#45 6 swallows/h and correlated well with net esophageal fluid flow. 'Bouts' of swallowing activity ( &#83 3 swallows/min) averaged 9 &#45 1 swallows/bout, lasted 1.8 &#45 1.4 min and accounted for 31 &#45 4% of the swallowed volume. Despite the absence of fetal electrocortical high-voltage/ low-voltage transitions, there was a 26.1 &#45 3.9-min interval between periods of swallowing bout activity. Conclusions: Preterm (0.75 gestation) ovine fetal volume swallowed (302 ml/day) and volume swallowed for body weight (175 ml/day per kg) was significantly less than that previously noted at 0.85 gestation (831 ml/day, 274 ml/day per kg, respectively; p < 0.05) although the rates of swallowing activity were similar. The presence of swallowing bout activity at periodic intervals, in the absence of electrocortical differentiation, suggests an intrinsic central pacemaker regulating preterm fetal neurobehavior.

Cellular Localization of Vascular Endothelial Growth Factor in Ovine Placenta and Fetal Membranes

To further understand the role of vascular endothelial growth factor (VEGF) in mediating angiogenesis and vascular permeability during development in the sheep placenta and fetal membranes, we examined the localization of VEGF mRNA and protein in placental, chorionic and amniotic tissues by in situ hybridization and immunohistochemistry in ovine fetuses at 62, 102 and 141 days gestation (term=150 days). In the placenta, VEGF mRNA expression and VEGF protein immunostaining were strong in cytotrophoblasts surrounding the villi. In addition, VEGF protein was localized in smooth muscle cells around fetal and maternal blood vessels and in the maternal epithelium. There was no apparent difference in placental VEGF mRNA or protein levels associated with advancing gestation. In the fetal membranes, VEGF mRNA was detected in the amniotic epithelium and the chorionic cytotrophoblastic cell layer. The intensity of the hybridization signals in both amnion and chorion appeared low at 62 days, moderate at 102 days and high at 141 days gestation. VEGF protein was detected in amniotic epithelium and chorionic cytotrophoblasts at all gestational ages studied. The increase in VEGF gene expression in fetal membranes as term approaches suggests that during fetal development VEGF may promote the vascularity and permeability of the microvessels which perfuse the fetal membranes, as well as permeability of the amniotic membrane itself. Thus VEGF may participate in the regulation of amniotic fluid volume.

Mechanism of arginine vasopressin suppression of ovine fetal lung fluid secretion: lack of V2-receptor effect.

Fetal lung liquid production is essential for in utero pulmonary development, and the resorption of lung liquid at birth facilitates neonatal transition. Lung liquid also contributes importantly to amniotic fluid volume. Factors that influence lung liquid production/resorption may, therefore, impact fetal pulmonary growth and development as well as amniotic fluid homeostasis. Arginine vasopressin (AVP) inhibits fetal lung liquid production and facilitates lung liquid resorption. In view of studies administering fetal AVP or the V2 receptor agonist, 1 desamino 8-D AVP (dDAVP) for the regulation of amniotic fluid volume, we sought to determine the impact of AVP V2 receptor stimulation on fetal lung liquid production. Eight near-term ovine fetuses (130 +/- 2 d; term = 145 d) were prepared with hind limb vascular catheters, a bladder catheter, and a tracheal catheter. Each ewe received vascular and amniotic catheters. After 5 days of recovery, the animals were studied for a 2-hr control period and for 2 hr following intravenous dDAVP injection (1 ng/kg). Lung liquid and urine flow rates and plasma electrolyte and osmolality composition were determined at 30-min intervals. To confirm fetal lung liquid suppression in response to AVP, two animals received an intravenous injection of AVP (2 ng/kg) 24 hr after the first experiment. dDAVP administration had no effect on lung liquid production (3.4 +/- 0.4 ml/kg/h), electrolyte concentrations, or osmolality. Fetal urine electrolyte concentrations and osmolality (154 +/- 24-295 +/- 22 mOsm/kg H2O) increased in response to dDAVP, whereas urine flow decreased (9.4 +/- 2.5-4.2 +/- 1.5 ml/kg/h). In the fetuses exposed to AVP, lung liquid production decreased (3.3-1.6 ml/kg/h). As AVP, although not dDAVP, inhibited fetal lung liquid production, these results indicate that AVP effects on lung liquid are likely mediated via the AVP V1 receptor, not the V2 receptor. The use of fetal administration of dDAVP for the regulation of urine production and amniotic fluid volume will not adversely impact lung liquid production.

Mechanism of arginine vasopressin suppression of ovine fetal lung fluid secretion: Lack of V2‐receptor effect

Fetal lung liquid production is essential for in utero pulmonary development, and the resorption of lung liquid at birth facilitates neonatal transition. Lung liquid also contributes importantly to amniotic fluid volume. Factors that influence lung liquid production/resorption may, therefore, impact fetal pulmonary growth and development as well as amniotic fluid homeostasis. Arginine vasopressin (AVP) inhibits fetal lung liquid production and facilitates lung liquid resorption. In view of studies administering fetal AVP or the V2 receptor agonist, 1 desamino 8-D AVP (dDAVP) for the regulation of amniotic fluid volume, we sought to determine the impact of AVP V2 receptor stimulation on fetal lung liquid production. Eight near-term ovine fetuses (130 ± 2 d; term = 145 d) were prepared with hind limb vascular catheters, a bladder catheter, and a tracheal catheter. Each ewe received vascular and amniotic catheters. After 5 days of recovery, the animals were studied for a 2-hr control period and for 2 hr following intravenous dDAVP injection (1 ng/kg). Lung liquid and urine flow rates and plasma electrolyte and osmolality composition were determined at 30-min intervals. To confirm fetal lung liquid suppression in response to AVP, two animals received an intravenous injection of AVP (2 ng/kg) 24 hr after the first experiment. dDAVP administration had no effect on lung liquid production (3.4 ± 0.4 ml/kg/h), electrolyte concentrations, or osmolality. Fetal urine electrolyte concentrations and osmolality (154 ± 24–295 ± 22 mOsm/kg H2O) increased in response to dDAVP, whereas urine flow decreased (9.4 ± 2.5–4.2 ± 1.5 ml/kg/h). In the fetuses exposed to AVP, lung liquid production decreased (3.3–1.6 ml/kg/h). As AVP, although not dDAVP, inhibited fetal lung liquid production, these results indicate that AVP effects on lung liquid are likely mediated via the AVP V1 receptor, not the V2 receptor. The use of fetal administration of dDAVP for the regulation of urine production and amniotic fluid volume will not adversely impact lung liquid production. J. Matern.–Fetal Med. 7:177–182, 1998. © 1998 Wiley-Liss, Inc.

Development of ingestive behavior

Swallowing represents a primary physiological function that provides for the ingestion of food and fluid. In precocial species, swallowing activity likely develops in utero to provide for a functional system during the neonatal period. The chronically instrumented ovine fetal preparation has provided the opportunity for recent advances in understanding the regulation of in utero swallowing activity. The near-term ovine fetus swallows fluid volumes (100-300 ml/kg) that are markedly greater, per body weight, than that of the adult (40-60 ml/kg). Spontaneous in utero swallowing and ingestive behavior contribute importantly to the regulation of amniotic fluid volume and composition, the acquisition and potential recirculation of solutes from the fetal environment, and the maturation of the fetal gastrointestinal tract. Fetal swallowing activity is influenced by fetal maturation, neurobehavioral state alterations, and the volume of amniotic fluid. Furthermore, intact dipsogenic mechanisms (osmolality, angiotensin II) have been demonstrated in the near-term ovine fetus. It remains unknown to what degree, if any, fetal swallowing may be influenced by nutrient appetite, salt appetite, or taste. Nevertheless, the development of dipsogenic and additional regulatory mechanisms for ingestive behavior occurs during fetal life and may be susceptible to changes in the pregnancy environment. This review describes what is currently known regarding the in utero development of ingestive behavior and the importance of this activity for fetal and perhaps ultimately adult fluid homeostasis.

Does gut atresia cause polyhydramnios?

Fetal gut atresia is variably associated with polyhydramnios. In order to determine which pregnancies will develop polyhydramnios, the case notes of 80 babies with gut atresia and stenosis were reviewed. Maternal polyhydramnios developed in all cases of pure oesophageal atresia (n = 8), all cases of Type III duodenal atresia (DA) with a non-bifid bile duct (n = 8), 80% of cases with type I DA (n = 10), and 24% of atresias of the small intestine (n = 34). Polyhydramnios did not develop in any case where there was not total obstruction except in 1 baby with DA and a bifid bile duct (BBD). These included stenosis of the oesophagus and duodenum (n = 17) and DA type III with a BBD (n = 3). These results support the role of fetal swallowing and fluid absorption by the fetal gastro-intestinal tract in the regulation of amniotic fluid volume.

Mechanism of arginine vasopressin suppression of ovine fetal lung fluid secretion: Lack of V2-Receptor effect

Fetal lung liquid production is essential for in utero pulmonary development, and the resorption of lung liquid at birth facilitates neonatal transition. Lung liquid also contributes importantly to amniotic fluid volume. Factors that influence lung liquid production/resorption may, therefore, impact fetal pulmonary growth and development as well as amniotic fluid homeostasis. Arginine vasopressin (AVP) inhibits fetal lung liquid production and facilitates lung liquid resorption. In view of studies administering fetal AVP or the V2 receptor agonist, 1 desamino 8-D AVP (dDAVP) for the regulation of amniotic fluid volume, we sought to determine the impact of AVP V2 receptor stimulation on fetal lung liquid production. Eight near-term ovine fetuses (130 ± 2 d; term = 145 d) were prepared with hind limb vascular catheters, a bladder catheter, and a tracheal catheter. Each ewe received vascular and amniotic catheters. After 5 days of recovery, the animals were studied for a 2-hr control period and for 2 hr following intravenous dDAVP injection (1 ng/kg). Lung liquid and urine flow rates and plasma electrolyte and osmolality composition were determined at 30-min intervals. To confirm fetal lung liquid suppression in response to AVP, two animals received an intravenous injection of AVP (2 ng/kg) 24 hr after the first experiment. dDAVP administration had no effect on lung liquid production (3.4 ± 0.4 ml/kg/h), electrolyte concentrations, or osmolality. Fetal urine electrolyte concentrations and osmolality (154 ± 24–295 ± 22 mOsm/kg H2O) increased in response to dDAVP, whereas urine flow decreased (9.4 ± 2.5–4.2 ± 1.5 ml/kg/h). In the fetuses exposed to AVP, lung liquid production decreased (3.3–1.6 ml/kg/h). As AVP, although not dDAVP, inhibited fetal lung liquid production, these results indicate that AVP effects on lung liquid are likely mediated via the AVP V1 receptor, not the V2 receptor. The use of fetal administration of dDAVP for the regulation of urine production and amniotic fluid volume will not adversely impact lung liquid production.

Anticholinergic suppression of ovine fetal swallowing activity

Objective: Fetal swallowing contributes importantly to amniotic fluid volume regulation as the primary route of fluid resorption, reaching 500 to 1000 ml/day near term. Near-term ovine fetal swallowing activity occurs predominantly during low-voltage electrocortical activity. In view of the potential to pharmacologically alter electrocortical activity, we hypothesized that fetal administration of a centrally acting cholinergic antagonist may be used to modulate fetal swallowing activity. To explore cholinergic modulation of swallowing activity, we examined fetal swallowing and electrocortical activity in response to central and peripheral cholinergic suppression by atropine sulfate. Study Design: Singleton ovine fetuses ( n = 6) were chronically prepared with vascular catheters and thyrohyoid, nuchal, and thoracic esophageal electromyogram and biparietal electrocortical electrodes. Swallowing and electrocortical activity were monitored for 2 hours before and after intravenous injection (1 ml of 0.15 mol/L sodium chloride) of atropine sulfate (1 mg/kg). On a subsequent day an identical study was performed with use off atropine methyl nitrate (3 mg/kg), an atropine analog that does not cross the blood-brain barrier. Results: Atropine sulfate decreased low-voltage electrocortical activity (56% ± 5% to 14% ± 4%), increased high-voltage electrocortical activity (40% ± 5% to 81% ± 5%), and did not change intermediate electrocortical activity (4% ± 1% to 5% ± 1%). Fetal swallowing activity decreased from 46 ± 12 to 12 ± 2 swallows per hour after atropine sulfate administration. Atropine methyl nitrate had no discernible effect on either fetal electrocortical or swallowing activity. Fetal arterial pressure, plasma osmolality, pH, Pco 2, and Po 2 did not change. Conclusions: Central cholinergic antagonism suppresses low-voltage fetal electrocortical and swallowing activity in the ovine fetus. Studies exploring spontaneous or induced fetal swallowing should consider the behavioral state of the fetus when conclusions are drawn about changes in the swallowing activity.

Vascular Endothelial Growth Factor: Possible Role in Fetal Development and Placental Function

Vascular endothelial growth factor (VEGF) is an endothelial cell mitogen with potent permeability properties. This growth factor exists in several isoforms; the most abundant form present in most tissues is VEGF165. The different isoforms exhibit differences in biologic function. During development, VEGF is expressed in multiple embryonic and fetal tissues, with the highest levels found in the lung, kidney, and heart. Vascular endothelial growth factor is also expressed in placental tissues and fetal membranes, and this expression increases with advancing gestation. In the fetal heart and placenta, VEGF expression is inducible by hypoxia. Two receptors, KDR and Flt-1, have been identified for VEGF. They are widely expressed in vascular endothelial cells and are also found in placental tissues where VEGF is localized. In humans, Flt-1 appears to be the predominant receptor, whereas in the cow and sheep, KDR is the major receptor expressed. The presence of VEGF and its receptors in placental tissues throughout gestation strongly suggests that VEGF plays an important role in the development and maintenance of placental vascular function during pregnancy. The localization of VEGF in fetal membranes and the fetal surface of the placenta raises the possibility that VEGF may be involved in the regulation of amniotic fluid volume and composition.

Physiology of amniotic fluid volume regulation.

Although there are fairly wide variations AFV normally undergoes characteristic changes across gestation in which it increases from 10-20 ml at 10 weeks gestation to average 800 ml at 24 weeks. Little change occurs from then until near term when AFV begins to decrease, and large decreases can occur in postterm pregnancies. Across gestation, 95% of AFVs are within the range of 1/2.57-2.57 times the gestational mean volume and 99% are within the range of 1/3.40-3.40 times the gestational mean. Although there are six pathways in which fluid and solutes can enter and/or leave the amniotic sac, there are only four primary pathways that contribute to AFV during late gestation. These include fetal urine and lung fluid secretion as the two primary sources of fluid, with fetal swallowing and intramembranous absorption as the two primary routes of amniotic water clearance. The intramembranous pathway also appears to be a primary source of amniotic solutes (e.g., sodium and chloride). Although fetal hypoxia has been widely believed to cause oligohydramnios, fetal hypoxic hypoxia and anemic hypoxia both appear to be associated with an increased AFV and polyhydramnios rather than oligohydramnios. It is speculated that the oligohydramnios associated with fetal hypoxia is caused by placental dysfunction in addition to the hypoxia.

The missing link in rhesus monkey amniotic fluid volume regulation: Intramembranous absorption

To investigate whether intramembranous absorption occurs in the rhesus monkey and its role in amniotic fluid (AF) volume regulation as a possible model for the human fetus. We studied five chronically catheterized rhesus monkey fetuses (Macaca mulatta) at 126 +/- 1 (standard error) days' gestation (term approximately 165 days) with ligated esophagi and catheterized tracheae. Samples (0.5 mL each) of fetal and maternal blood and amniotic and lung fluid were collected at 0, 15, 30, 60, 120, 180, and 240 minutes after injection of 0.1 mCi (3 mL) of technetium-99m (Tc-99m) into the amniotic cavity. In spite of esophageal ligation, there was a rapid absorption of the Tc-99m into the fetal circulation within 15 minutes of injection. The maternal Tc-99m activity increased in parallel to fetal activity but remained lower. The fetal lung fluid Tc-99m activity increased more slowly but was equivalent to the fetal circulating level by 4 hours. These results suggest that intramembranous absorption occurs and may play an important role in rhesus AF volume regulation and composition. Furthermore, this animal model, which closely resembles the human, may provide valuable insight into abnormalities of human AF volume regulation.

The Missing Link in Rhesus Monkey Amniotic Fluid Volume Regulation

In Brief Objective To investigate whether intramembranous absorption occurs in the rhesus monkey and its role in amniotic fluid (AF) volume regulation as a possible model for the human fetus. Materials and Methods We studied five chronically catheterized rhesus monkey fetuses (Macaca mulatto) at 126 ± 1 (standard error) days' gestation (term approximately 165 days) with ligated esophagi and catheterized tracheae. Samples (0.5 mL each) of fetal and maternal blood and amniotic and lung fluid were collected at 0,15,30, 60,120,180, and 240 minutes after injection of 0.1 mCi (3 mL) of technetium-99m (Tc-99m) into the amniotic cavity. Results In spite of esophageal ligation, there was a rapid absorption of the Tc-99m into the fetal circulation within 15 minutes of injection. The maternal Tc-99m activity increased in parallel to fetal activity but remained lower. The fetal lung fluid Tc-99m activity increased more slowly but was equivalent to the fetal circulating level by 4 hours. Conclusions These results suggest that intramembranous absorption occurs and may play an important role in rhesus AF volume regulation and composition. Furthermore, this animal model, which closely resembles the human, may provide valuable insight into abnormalities of human AF volume regulation. Technetium-99m injected into the moneky amniotic cavity is absorbed rapidly into the fetal circulation despite fetal esophageal ligation, suggesting the presence of intramembranous absorption.

The missing-link in primate amniotic fluid volume regulation: intra-membranous absorption.

The missing-link in primate amniotic fluid volume regulation: intra-membranous absorption.

Progress toward understanding the regulation of amniotic fluid volume: Water and solute fluxes in and through the fetal membranes

Presently the regulation of amniotic fluid volume is poorly understood. There are multiple reasons for this: (1) the volume regulatory mechanisms are complex because there are multiple pathways over which water and solute exchanges occur simultaneously; (2) the volume regulatory mechanisms differ dramatically in early compared to late gestation; (3) diffusional water movement across the membranes has sometimes been confused with net water movement (which is orders of magnitude less); and (4) there appears to be extensive and rapid exchanges between amniotic fluid and fetal blood which occur on the fetal surface of the placenta and within the vascularized fetal membranes (for those species in which the membranes are vascularized) which have not been fully recognized. Knowledge of the direct exchange between amniotic fluid and fetal blood is undergoing significant expansion. Over the past S-10 years, there have been major advances which show that reabsorption of amniotic water and solutes into fetal blood within the membranes and into the fetal surface of the placenta is an important contributor to amniotic homeostasis. Further, there is an accumulating body of data characterizing exchange (i.e. the lack of) between amniotic fluid and the maternal uterine wall. Thus, the present review attempts to clarify these issues by integrating available information regarding fluxes within and across the fetal membranes. Since a large majority of the new data is from the last half of pregnancy, only data derived during that time period will be considered below. Further, although data from humans, monkeys and sheep are discussed, it is important to recognize that a large majority of the data generated during the past few years has been derived from the chronically catheterized fetal sheep. Several of these recent studies have used the sheep model where the urachus was ligated or fetal urine was diverted into the amniotic sac so that no allantoic fluid was present. This represents an improved animal model in comparison with the normal ovine fetus for studying the dynamics and control of amniotic fluid volume and composition.