Maternal plasma hypo-osmolality: effects on spontaneous and stimulated ovine fetal swallowing.

Fetal swallowing is a major route of amniotic fluid resorption, and thus swallowing activity may alter amniotic fluid volume. Near-term ovine fetal swallowing increases in response to plasma and/or cerebrospinal fluid hypertonicity. As maternal hydration status alters amniotic fluid volume, we hypothesized that maternal plasma hypotonicity may alter fetal swallowing activity. Pregnant ewes (130 +/- 1 d; n = 6) were chronically prepared with maternal and fetal vascular catheters, a fetal esophageal flow probe, and fetal thyrohyoid and nuchal and thoracic esophagus electromyogram electrodes. Spontaneous fetal swallowing and hypertonic saline thresholds for stimulated swallowing were determined prior to and following maternal hypotonicity induced with water loading and intravenous DDAVP (arginine vasopressin V2 agonist). Fetal swallowing thresholds were determined with intracarotid injections (0.15 ml/kg) of increasing sodium chloride concentrations (0.15-1.2 M) at 2-min intervals. Maternal DDAVP infusion significantly decreased mean (+/-SEM) maternal and fetal plasma osmolalities (298 +/- 2-284 +/- 3; 295 +/- 2-278 +/- 3 mOsm/kg, respectively) and sodium concentrations (147.3 +/- 0.4-137.5 +/- 0.9; 142.7 +/- 0.8-133.5 +/- 1.0 mEq/l, respectively), suppressed spontaneous swallowing activity and volume (1.1 +/- 0.2-0.6 +/- 0.1 swallows/min; 0.7 +/- 0.2-0.5 +/- 0.1 ml/min, respectively) and significantly increased the osmotic threshold for swallowing stimulation (0.77 +/- 0.08-1.03 +/- 0.09 M NaCl). We conclude that: (1) maternal, and thus fetal, plasma hypotonicity results in suppression of spontaneous fetal swallowing activity and a decrease in volume swallowed, suggesting that spontaneous fetal ingestive behavior results, in part, from tonic dipsogenic stimulation, and (2) under hypotonic conditions, the intracarotid NaCl injection concentration for swallowing stimulation increases. These results suggest that the reset (lower) maternal plasma osmolality during human pregnancy may serve to minimize fetal ingestive and perhaps arginine vasopressin-mediated antidiuretic responses to acute maternal hypertonicity.

Ovine fetal laryngeal chemoreflex thresholds and respiratory effects.

In newborn infants, laryngeal contact with solutions of low chloride concentration or pH evokes swallowing, laryngeal adduction, and respiratory inhibition (laryngeal chemoreflex). To determine whether the laryngeal chemoreflex is present during fetal life and its effect on fetal respiratory activity, eight time-bred ewes (128 +/- 2 days) were prepared with fetal electrocortical diaphragm and esophageal electrodes and a nasopharyngeal catheter. After a 60-minute control period, increasing volumes (0.1 to 1.0 ml/kg) of 0.15 mol/L NaCl or distilled water (0.05 to 1.0 ml/kg) and decreasing concentrations of NaCl (0.15 to 0.02 mol/L) at a fixed volume (0.3 ml/kg) were sequentially administered through the nasopharyngeal catheter (38 degrees C). The minimum water volume that stimulated swallowing was significantly less than the minimum 0.15 mol/L NaCl volume (0.10 +/- 0.02 vs. 0.70 +/- 0.05 ml/kg). The maximum NaCl concentration that stimulated swallowing was 0.04 +/- 0.01 mol/L During the control period, respiratory activity averaged 14.6 +/- 0.7 breaths/minute and did not change during absent swallow responses or isotonic saline-induced swallows. However, respiratory activity significantly decreased during water (4.7 +/- 0.6 breaths/minute) and hypotonic saline-induced swallow responses (3.7 +/- 0.7 breaths/minute). Fetal electrocortical activity did not change during absent or stimulated swallows. We conclude that laryngeal water or hypotonic saline solution may stimulate fetal swallowing and suppress fetal respiratory activity, similar to the newborn laryngeal chemoreflex. We speculate that an exaggeration of the laryngeal chemoreflex apnea response in the newborn may predispose to sudden infant death syndrome.

1-Deamino-[8-D-arginine] vasopressin-induced maternal plasma hypoosmolality increases ovine amniotic fluid volume.

OBJECTIVE: Maternal 1-deamino-[8-D-arginine] vasopressin (a selective antidiuretic agonist) and oral water loading decrease maternal and fetal plasma osmolality and markedly increase fetal urine flow in sheep over 24 hours. We hypothesized that a sustained reduction in plasma osmolality would increase the amniotic fluid volume. STUDY DESIGN: Pregnant sheep (130 +/- 1 days, n = 6) were given oral water loading (2000 ml) and intravenous 1-deamino-[8-D-arginine] vasopressin (20 microgram injection followed by 4 micrograms/hr) to induce maternal urinary antidiuresis and to maintain a maternal plasma osmolality decrease of approximately 20 mOsm/kg for 48 hours. Fetal plasma osmolality, electrolytes, atrial natriuretic factor, and arginine vasopressin levels, fetal swallowing activity, and fetal urine flow and composition were measured each study day, and amniotic fluid volume was measured by tracer dilution before and at 48 hours of plasma hypoosmolality. RESULTS: In response to 1-deamino-[8-D-arginine] vasopressin and water loading, maternal plasma osmolality decreased (298 +/- 1 to 275 +/- 3 mOsm/kg), mirrored by fetal plasma hypoosmolality (293 +/-1 to 271 +/- 3 mOsm/kg). Fetal urine osmolality (155 +/- 16 to 117 +/- 10 mOsm/kg) and amniotic fluid osmolality significantly decreased (281 +/- 3 to 269 +/- 5 mOsm/kg). Fetal urine flow rate significantly increased (0.33 +/- 0.06 to 0.51 +/- 0.04 ml/min), whereas fetal swallowing activity decreased (56 +/- 7 to 33 +/-5 swallows per hour). In response to alterations in fetal fluid dynamics, amniotic fluid volume significantly increased from 493 +/- 80 to 839 +/- 160 ml. CONCLUSION: Chronic maternal 1-deamino-[8-D-arginine vasopressin-induced plasma hypoosmolality induces a sustained reduction in fetal swallowing activity and an increase in fetal urine production, which results in a marked increase in amniotic fluid volume. Use of maternal 1-deamino-[8-D-arginine] vasopressin and oral fluid loading has the potential for manipulation of amniotic fluid volume homeostasis and may prove useful in the treatment of oligohydramnios.

Development of concentration thresholds for fetal swallowing and arginine vasopressin stimulation in the preterm ovine fetus.

We previously demonstrated that fetal swallowing was stimulated by intracarotid saline injections in the near term (130 +/- 1 days gestation) ovine fetus. In order to examine the ontogeny of the regulation of thirst-mediated swallowing behavior and arginine vasopressin (AVP) secretion, the present study examined fetal swallowing and AVP secretory responses to hypertonic NaCl and sucrose in preterm (115 days gestation) ovine fetuses. Five fetuses were chronically prepared with thyrohyoid, nuchal and thoracic esophagus, and diaphragm electromyogram (EMG) electrodes, an esophageal flow probe, and vascular catheters. Fetuses received intracarotid injections (0.15 ml/kg) of increasing concentrations of NaCl (0.15-2.55 M), administered at 2-min intervals. The threshold saline concentration for swallowing was defined as the minimum NaCl dose eliciting swallow responses (within 20 s) following 4 of 5 injections at each dose. In 2 fetuses, the threshold dose was 0.75 M (confirmed by 4 of 5 injections resulting in swallows at the next higher dose). In the other 3 animals, no swallowing threshold was observed (i.e., no dose through 2.55 M elicited swallows to 4 of 5 injections). There were no significant increases in AVP secretion. On a subsequent day, equimolar sucrose injections did not affect fetal swallowing activity or AVP secretion. Together with previous reports, these studies indicate that plasma osmolality dipsogenic mechanisms become functional in some ovine fetuses by 115 days gestation, while all fetuses exhibit intact osmotic dipsogenic responses at 130 days gestation. These results further suggest an acute process of development of dipsogenic responsiveness, rather than by gradually increasing sensitivity to osmotic stimuli.

Intraamniotic deamino(D-Arg8)-vasopressin: prolonged effects on ovine fetal urine flow and swallowing.

OBJECTIVE: Fetal urine is the main source of amniotic fluid, and alterations in urine production have an impact on amniotic fluid dynamics. Intraamniotic arginine vasopressin is absorbed into fetal plasma, but fetal antidiuretic responses may be obscured by vasoconstrictor (V1 receptor) actions of arginine vasopressin. We determined the effects of intraamniotic administration of a specific V2 receptor agonist, deamino(D-Arg8)-vasopressin, on fetal plasma arginine vasopressin immunoreactivity, fetal urine output, and swallowing. STUDY DESIGN: Six chronically prepared pregnant ewes (129 +/- 1 days' gestation) received a single deamino(D-Arg8) (50 micrograms injection into the amniotic fluid with subsequent measurement of fetal plasma arginine vasopressin immunoreactivity, fetal urine flow and osmolality, and fetal swallowing. RESULTS: Within 30 minutes of deamino(D-Arg8)-vasopressin administration, fetal plasma arginine vasopressin immunoreactivity (4.5 +/- 0.5 to 76.4 +/- 21.9 pg/ml) and urine osmolality (135 +/- 13 to 285 +/- 43 mOsm/kg H2O) significantly increased and urine flow decreased (0.18 +/- 0.01 to 0.05 +/- 0.01 ml/kg per minute). After 48 hours urine flow remained reduced (0.08 +/- 0.03 mg/kg per minute) and osmolality remained elevated (399 +/- 18 mOsm/kg H2O). There were no changes in fetal swallowing, systolic (64 +/- 3 mm Hg) or diastolic (42 +/- 3 mm Hg) blood pressure, or heart rate (169 +/- 6 beats/min). CONCLUSIONS: Intraamniotic deamino(D-Arg8)-vasopressin administration evokes persistent fetal antidiuresis in the absence of arginine vasopressin-induced fetal cardiovascular effects or changes in fetal swallowed volume, indicating its suitability as a potential therapeutic agent in amniotic fluid volume disorders such as polyhydramnios.

Cocaine stimulation of ovine fetal swallowing.

Cocaine acts to block reuptake of neurotransmitters, resulting in elevated intra-synaptic norepinephrine levels. As monoaminergic systems may contribute to central regulation of swallowing motor activity as well as dipsogenic responses, and cocaine is known to alter fetal behavior, we examined the effect of intravenous cocaine on fetal swallowing. Six ovine fetuses (130 +/- 1 days) were chronically prepared with esophageal electrodes and an esophageal flow probe. Following a 1-h control period, fetuses received an intravenous injection of 1.0 mg/kg cocaine over 30 s. Fetal blood samples were withdrawn at timed intervals and fetal swallowing activity was monitored for 180 min after the cocaine bolus. Basal fetal swallowing activity during the control period was 0.6 +/- 0.1 swallows/min, with esophageal flow of 0.3 +/- 0.1 ml/min. At 10 min following cocaine, fetal swallowing rate increased to 1.4 +/- 0.4 swallows/min though esophageal flow did not change significantly (0.4 +/- 0.2 ml/min). Swallowing rate then returned to basal levels. Fetal heart rate increased in response to cocaine injection (149 +/- 14 to 174 +/- 24 bts/min at 180 min) though there was no change in fetal mean blood pressure. These results demonstrate acute stimulation of fetal swallowing activity in response to intravenous cocaine injection. Together with previous reports, these data further illustrate the diversity of fetal behavioral effects in response to in utero cocaine exposure.

Ovine fetal swallowing response to intracerebroventricular hypertonic saline.

In the near-term ovine fetus, systemic hyperosmolality stimulates dipsogenic responses. Putative systemic dipsogens (hypertonicity, angiotensin II) may initiate responses by stimulation of select cerebral circumventricular nuclei lacking a blood-brain barrier. To investigate whether central osmotic-dipsogenic mechanisms are functional in utero, fetal swallowing responses to intracerebroventricular (i.c.v.) hypertonic saline were examined. Five pregnant ewes with singleton fetuses (128 +/- 1 days gestation) were prepared with fetal lateral cerebral ventricle and vascular catheters, electrocortical (ECoG) electrodes, and electromyogram wires on the fetal thyrohyoid muscle, nuchal and thoracic esophagus, and diaphragm and studied for a minimum of 5 days postoperatively. After a 2-h basal period, fetuses received an i.c.v. infusion of artificial cerebrospinal fluid followed by successive 30-min infusions of hypertonic NaCl in artificial cerebrospinal fluid (500 and 700 mosmol/kgH2O). In response to the i.c.v. hypertonic NaCl infusions, fetal swallowing significantly increased (1.4 +/- 0.4 to 3.9 +/- 1.4 and 2.9 +/- 0.5 swallows/min low-voltage ECoG, respectively). Plasma arginine vasopressin levels increased, although the change was not statistically significant (9.1 to 24.2 pg/ml; P = 0.07), and there was no change in fetal plasma osmolality, sodium concentration, or ECoG activity. Together with previous studies, these results indicate that both central and systemic osmotic dipsogenic mechanisms are functional in utero.

DDAVP-induced maternal hyposmolality increases ovine fetal urine flow.

Fetal urine flow is influenced by fetal intravascular volume, glomerular filtration rate, tubular reabsorption, and fluid regulatory hormones. As maternal-to-fetal fluid transfer is dependent on hydrostatic and osmotic gradients, we postulated that a chronic decrease in maternal plasma osmolality would promote transplacental fluid transfer and increase fetal urine flow. Six pregnant ewes and singleton fetuses (131 +/- 2 days; term = 150 days) received bladder and hindlimb arterial and venous catheters. After 5 days, plasma and urine composition, urine flow rate (Uvol), and plasma arginine vasopressin (AVP) levels were measured during a 2-h control period. At 2 h, tap water (2 liter, 38 degrees C) was administered to the ewe. At 3 h, ewes received a 20-micrograms bolus of 1-desamino-[D-Arg8]vasopressin (DDAVP), followed by continuous infusion (4 micrograms/h). In response to water loading, maternal urine osmolality decreased (761 +/- 158 to 339 +/- 13 mosmol/kgH2O), and Uvol increased. After DDAVP, maternal urine osmolality increased (1,270 +/- 89 mosmol/kgH2O), and Uvol, hematocrit, plasma osmolality (304 +/- 1 to 284 +/- 4 mosmol/kgH2O), and protein concentration decreased. Five hours after maternal DDAVP infusion, fetal plasma osmolality decreased (300 +/- 1 to 281 +/- 3 mosmol/kgH2O), and Uvol increased (0.4 +/- 0.1 to 1.3 +/- 0.2 ml/min) and remained elevated at 24 h. There was no change in fetal plasma DDAVP (immunoreactive AVP) levels or fetal urine osmolality. Controlled changes in maternal plasma osmolality may prove useful in modulating fetal urine flow and, ultimately, amniotic fluid volume.

Central angiotensin II stimulation of ovine fetal swallowing.

In the late-gestation ovine fetus, spontaneous swallowing occurs primarily during fetal low-voltage electrocortical (ECoG) activity in association with fetal breathing movements. Fetal swallowing activity may be stimulated in response to systemic or carotid plasma hyperosmolality, although not to increased plasma angiotensin II (ANG II) levels. In view of the potent dipsogenic effects of central, but not peripheral, ANG II in adult sheep, the present study investigated the effect of intracerebroventricular (ICV) ANG II on fetal swallowing activity. Six ovine fetuses (127 +/- 1 days) were chronically prepared with electromyogram and cortical electrodes and with vascular and lateral ventricle catheters. After a 2-h control period, fetuses received ICV injections of artificial cerebrospinal fluid and increasing concentrations of ANG II (0.1, 1.0, 10, 100, and 500 ng/kg). Fetal ECoG activity did not change, although fetal swallowing activity significantly increased in response to the 100- and 500- ng/kg ANG II doses (1.20 +/- 0.14 to 3.34 +/- 0.59 and 3.46 +/- 0.81 swallows/min of low-voltage ECoG, respectively). In response to the highest ANG II dose, fetal plasma arginine vasopressin levels significantly increased (5.7 +/- 1.2 to 17.2 +/- 4.6 pg/ml). ICV ANG II stimulation of fetal swallowing and arginine vasopressin secretion demonstrates that central ANG II dipsogenic mechanisms are intact by 0.9 of ovine gestation.

Ovine fetal swallowing and renal responses to oligohydramnios.

Amniotic fluid (AF) volume regulation is dependent on a balance between fluid production and fluid resorption. We examined the effects of reduced AF volume on AF production by fetal urine and resorption by fetal swallowing and the response of these parameters to AF volume replacement. Eight time-dated pregnant ewes (125 +/- 1 days gestation) were studied before (day 1) and after (day 3) AF and fetal urine drainage. Drainage resulted in a significant decrease in AF volume (415 +/- 89 to 157 +/- 36 ml). Fetal urine osmolality increased (139 +/- 10 to 286 +/- 33 mosmol/kgH2O), while urine flow did not change significantly (0.31 +/- 0.04 to 0.23 +/- 0.06 ml/min), resulting in nonsignificant increases in osmolar, sodium, and chloride excretions. Fetal electromyographic swallowing activity decreased 30% (1.0 +/- 0.1 to 0.7 +/- 0.1 swallows/min; P < 0.05), while net esophageal flow decreased 74% (0.31 +/- 0.12 to 0.07 +/- 0.04 ml/min; P < 0.05). On day 4, 0.15 M NaCl (500 ml; 37 degrees C) was administered into the AF over 30 min. During the 2 h after reinfusion, urine flow (0.29 +/- 0.07 to 0.40 +/- 0.09 ml/min) and osmolar sodium and chloride excretion significantly increased, though fetal swallowing activity and esophageal flow did not change. Thus the ovine fetus responded to reduced AF volume by maintaining AF production and decreasing AF resorption. In response to AF replacement, urine flow increased while fetal swallowing activity did not change, consistent with an intramembranous pathway for fetal AF resorption.

Ontogeny of ovine fetal renal atrial natriuretic factor receptors.

Ovine fetal renal responses to ANF decrease during the last third of gestation, although circulating fetal plasma atrial natriuretic factor (ANF) levels are higher than in the maternal circulation, and do not change with gestation. This study examined whether previously reported maturational changes in fetal renal responses to ANF are due to changes in renal ANF receptor numbers and/or affinity during gestation. ANF receptor numbers (Bmax) and dissociation constants (Kd) were measured in isolated renal glomeruli from early (95 and 110 day; mean 103 +/- 2) and late gestation (131 and 145 day; mean 138 +/- 2) fetal and maternal sheep. Fetal renal ANF receptor Bmax values significantly increased between 103 and 138 days gestation (13 +/- 3 to 29 +/- 4 fmol/mg protein) but were significantly lower than maternal values (60 +/- 13 fmol/mg protein). Fetal ANF receptor Kd values also increased significantly (245 +/- 34 to 370 +/- 36 pM), with early gestation values significantly lower than maternal values (470 +/- 69 pM). Thus, the blunted fetal renal response to ANF in late as compared to early gestation is not due to reduced ANF receptor numbers. Rather, an increased proportion of ANF clearance receptors, reduced post-receptor function and/or altered intrarenal hemodynamics may contribute.

Assessments of fetal swallowed volume: tracer disappearance versus esophageal flow.

OBJECTIVE: We sought to compare fetal swallowed volume determinations simultaneously by two techniques--amniotic fluid (AF) tracer disappearance and esophageal flow probe measurements. METHODS: Six ovine fetuses (129 +/- 1 days) were chronically prepared with a thoracic esophageal flow probe and vascular and two AF catheters. 125I-labeled albumin was injected into the AF cavity, and samples were withdrawn at timed intervals for 8 hours. The AF volume was calculated by time-0 extrapolation of the semilog of 125I disappearance. Tracer-determined swallowed volume was calculated as the product of AF volume and the slope of isotope disappearance. Flow probe measurement of swallowed volume was determined by computer integration of calibrated flow probe velocity recordings. RESULTS: The AF volume averaged 805 +/- 168 mL. The isotope disappearance rate from the AF was 2.8 +/- 0.4%/hour. Average tracer-determined swallowed volume (547 +/- 113 mL/day) was greater than flow probe volume (366 +/- 81 mL/day), although these values were not significantly different. However, when corrected for estimated swallowed lung fluid, tracer-determined volume was significantly greater than flow probe volume (P < .05). CONCLUSION: Ovine fetal swallowed volume determinations by AF tracer techniques are greater than those determined by esophageal flow probe measurements.

Fetal endocrine and renal responses to in utero ventilation and umbilical cord occlusion.

OBJECTIVE: Fetal to neonatal transition involves a myriad of endocrine and renal adaptive changes triggered by multiple simultaneous stimuli. We examined the extent to which ventilation and umbilical cord occlusion have an impact on the many endocrine and renal function changes in fetal sheep at 133 +/- 1 day of gestation. STUDY DESIGN: Nine fetuses were chronically prepared with an endotracheal tube, vascular and bladder catheters, and an inflatable umbilical cord occluder. After a 2-hour control period fetuses were treated with commercially prepared surfactant and ventilated with 100% oxygen. One hour after the onset of stable ventilation the umbilical cord was occluded and the animals were monitored for 3-hours. RESULTS: In response to ventilation fetal arterial PO2 increased (18 +/- 1 to 86 +/- 29 mm Hg) and remained significantly above control values after cord occlusion. Fetal arterial pH, hematocrit, and mean arterial pressure and heart rate did not change during the study. Ventilation alone evoked significant increases in epinephrine and norepinephrine concentrations. Renal responses to ventilation included significant decreases in urine flow rate, fractional sodium excretion, and fractional water excretion. Neither ventilation nor cord occlusion resulted in significant changes in plasma concentrations of atrial natriuretic factor, arginine vasopressin, and angiotensin II or in glomerular filtration rate, urine osmolality, free water, and osmolar clearances. CONCLUSIONS: (1) Mechanical ventilation and oxygenation alone increase circulating fetal catecholamine levels. (2) The addition of umbilical cord ligation has minimal impact on fetal endocrine and renal adaptive responses.

Maternal/fetal dehydration: prolonged effects and responses to oral rehydration.

Dehydration induces marked alterations in maternal-fetal fluid homeostasis and accompanying fetal endocrine responses. We sought to determine if the increase in fetal plasma arginine vasopressin (AVP) levels during maternal dehydration is mediated by fetal plasma hypovolemia in addition to hyperosmolality and to examine maternal and fetal plasma atrial natriuretic factor (ANF) responses to maternal dehydration and oral rehydration. Seven pregnant ewes (127 +/- 1 day) were water deprived for 72-96 h, and five of these were orally rehydrated. Dehydration induced significant increases in maternal plasma osmolality (pOSM) (300 +/- 2 to 325 +/- 8 mosmol/kg) and AVP (3.0 +/- 0.4 to 18.9 +/- 4.0 pg/ml), and decreases in plasma ANF levels (28.1 +/- 3.1 to 19.7 +/- 3.1 pg/ml). Fetal pOSM (293 +/- 3 to 314 +/- 4 mosmol/kg), AVP (2.5 +/- 0.6 to 8.1 +/- 4.8 pg/ml), and urinary fractional sodium excretion increased significantly, whereas plasma ANF and fetal blood volume did not change. After maternal water access maternal plasma AVP decreased rapidly in comparison to the gradual decrease in maternal pOSM. Fetal plasma AVP levels did not change significantly and fetal pOSM decreased more slowly than maternal pOSM. Fetal plasma ANF increased in association with increased urine flow and glomerular filtration rate after maternal rehydration. These data indicate marked differences in fetal and maternal plasma ANF and AVP responses with dehydration-induced increases in fetal plasma AVP being secondary to plasma hyperosmolality, rather than hypovolemia. Rapid suppression of maternal plasma AVP may contribute to the slower equilibration of fetal pOSM during oral, as compared with intravenous, maternal rehydration.

Dehydration increases the renal response to atrial natriuretic peptide in fetal sheep.

OBJECTIVE: In sheep, maternal water deprivation results in urinary natriuresis in spite of suppression of plasma atrial natriuretic factor levels. Near-term fetal sheep also have a urinary natriuresis without change in plasma atrial natriuretic factor during maternal dehydration. This study was designed to explore the role of plasma atrial natriuretic factor levels in fetal dehydration-natriuresis. STUDY DESIGN: Eight chronically instrumented preterm (113 +/- 1 days) ovine fetuses received two atrial natriuretic factor infusions (3 and 15 ng/kg/min) in a euhydrated state and after 48 +/- 1 hours of maternal water deprivation. RESULTS: Dehydration significantly increased maternal plasma osmolality (302 +/- 2 to 313 +/- 2 mOsm/kg water), sodium (148.1 +/- 0.8 to 154.3 +/- 0.4 mEq/L), chloride (112.4 +/- 0.6 to 116.8 +/- 0.9 mEq/L), and arginine vasopressin (4.2 +/- 1.2 to 23.0 +/- 4.0 pg/ml) and significantly decreased plasma atrial natriuretic factor (36 +/- 6 to 19 +/- 4 pg/ml) concentrations. Fetal plasma osmolality (296 +/- 1 to 308 +/- 2 mOsm/kg), atrial natriuretic factor (128 +/- 16 to 241 +/- 36 pg/ml), and arginine vasopressin (3.5 +/- 0.8 to 12.3 +/- 4.8 pg/ml) concentrations and urine osmolality (170 +/- 10 to 253 +/- 10 mOsm/kg), osmolar clearance (0.80 +/- 0.02 to 0.14 +/- 0.02 ml/kg/min), and fractional sodium excretion (3.3% +/- 1.7% to 8.5% +/- 2.1%) increased significantly with dehydration, whereas the plasma atrial natriuretic factor clearance decreased from 127 +/- 27 to 63 +/- 10 ml/kg/min. Dehydration had no effect on fetal hematocrit, vascular pressures, glomerular filtration rate, urine flow, or free water clearance. In euhydrated fetuses plasma atrial natriuretic factor increased from 128 +/- 16 to 287 +/- 46 pg/ml with sequential atrial natriuretic factor infusion, and no significant increases were observed in urine flow, fractional sodium excretion, and glomerular filtration rate. In contrast, atrial natriuretic factor infusion to dehydrated fetuses significantly increased urine flow (0.17 +/- 0.03 to 0.32 +/- 0.07 ml/kg/min), osmolar clearance (0.14 +/- 0.02 to 0.28 +/- 0.06 ml/kg/min), and fractional sodium excretion (8.5% +/- 2.1% to 14.8% +/- 4.0%). CONCLUSION: These results demonstrate that in the fetus at 113 days' gestation plasma atrial natriuretic factor levels increase with dehydration, probably a result of decreased plasma atrial natriuretic factor clearance, and the fetal renal responsiveness to atrial natriuretic factor infusion increases during maternal dehydration.

Ovine maternal and fetal renal vasopressin receptor response to maternal dehydration.

OBJECTIVE: Arginine vasopressin secretion increases in response to increased plasma osmolality or hypovolemia. Dehydration-induced increases in plasma arginine vasopressin levels have been shown to down-regulate arginine vasopressin V2 receptors in adult rat kidneys. Our study determined ovine maternal and fetal renal arginine vasopressin receptor characteristics and receptor response to maternal dehydration. STUDY DESIGN: Eight pregnant ewes (113 +/- 1 days) were dehydrated for 72 hours; eight animals served as controls. Renal medullary tissue was isolated from maternal and fetal kidneys, and arginine vasopressin receptor characteristics determined with saturation and competition assays using tritiated arginine vasopressin, arginine vasopressin, and arginine vasopressin analogs. RESULTS: Euhydrated maternal and fetal renal medullary arginine vasopressin receptor dissociation constant (3.0 +/- 0.3 and 1.9 +/- 0.3 nmol/L) and maximal binding capacity (149 +/- 15 and 111 +/- 33 fmol/mg protein) values were similar. Pharmacologic profiles with selective agonists indicated a predominance of V2 receptors. Dehydration significantly increased maternal and fetal plasma osmolalities (304 +/- 2 to 320 +/- 2; 296 +/- 1 to 319 +/- 3 mOsm/kg water, respectively) and arginine vasopressin levels (3.8 +/- 1.4 to 29.3 +/- 4.6; 4.4 +/- 1.0 to 16.9 +/- 5.0 pg/ml, respectively) but had no effect on arginine vasopressin receptor binding. CONCLUSION: Specific, saturable, single-site tritiated arginine vasopressin binding is present in ovine maternal and fetal renal medullary membranes. Ovine maternal and fetal renal arginine vasopressin receptors do not down-regulate in response to dehydration-induced elevations in plasma arginine vasopressin levels.

Differential ontogeny of in vitro vascular responses to three categories of calcium channel antagonists in rats.

We examined the ontogeny of relaxation responses to three categories of calcium channel antagonists, represented by verapamil, diltiazem, and nifedipine, for both potential-operated (KCl-mediated) and receptor-operated channels [norepinephrine (NE)-mediated] in rat thoracic aorta. Aortic rings from 2- to 3-d, 1-wk, and 12-wk-old Sprague Dawley rats were mounted in an organ bath, bathed in Krebs' solution, and connected to a force-displacement transducer to measure isometric tension. Endothelium intact vessels at optimal passive force were exposed to a single ED50 of isotonic KCl or NE, equilibrium contraction was measured, then vessels were washed and exposed for 30 min to 1 microM verapamil, 1 microM diltiazem, or 0.1 microM nifedipine, followed by another dose of KCl or NE. Verapamil and diltiazem demonstrated significant (p less than 0.05) age-related increases in effectiveness for blocking KCl-mediated contraction [(% reduction of control contraction +/- SEM) (Verapamil: 2-3 d, 67.7 +/- 4.2; 1 wk, 72.5 +/- 1.8; 12 wk, 89.5 +/- 1.0. Diltiazem: 2-3 d, 64.6 +/- 2.9; 1 wk, 73.5 +/- 3.0; 12 wk, 83.1 +/- 1.8]. Nifedipine was equally effective at all ages: 2-3 d, 85.6 +/- 1.3; 1 wk, 90.0 +/- 1.6; and 12 wk, 91.3 +/- 1.4. Verapamil and diltiazem also showed significant age-related increases in effectiveness for blocking NE-mediated contraction (Verapamil: 2-3 d, 6.2 +/- 3.9; 1 wk, 28.0 +/- 4.8; 12 wk, 44.1 +/- 6.0. Diltiazem: 2-3 d, 8.0 +/- 3.1; 1 wk, 20.5 +/- 3.9; 12 wk, 46.5 +/- 4.8).(ABSTRACT TRUNCATED AT 250 WORDS)

Developmental changes in sodium nitroprusside and atrial natriuretic factor mediated relaxation in the guinea pig aorta.

Sodium nitroprusside (SNP), a nonreceptor mediated stimulant of soluble guanylate cyclase, and atrial natriuretic factor, a receptor-dependent stimulator of particulate guanylate cyclase, mediate relaxation responses by increasing intracellular cGMP. This in vitro study was designed to compare the ontogeny of relaxation responses to SNP and atrial natriuretic factor in the guinea pig thoracic aorta. Aortic rings from fetuses at 55-60 d gestation (term = 68 d), 1- to 3-d-old newborn, and 12-wk-old adult Hartley guinea pigs were mounted in an organ bath, bathed in Kreb's solution, and connected to a force-displacement transducer to measure isometric tension. Relaxation responses to SNP and atriopeptin III were studied with the vessels at optimal resting tension and after preconstriction with an EC85 concentration of norepinephrine. SNP-mediated relaxation showed a significant increase in sensitivity with development among the three age groups (p less than 0.05). Methylene blue, an inhibitor of soluble guanylate cyclase, produced no inhibition of relaxation to SNP in fetal aortae, significantly decreased responses along the straight portion of the concentration-response curve in newborn aortae (p less than 0.05), and significantly shifted the concentration-response curve to the right (p less than 0.05) in adult aortae; but did not prevent vessels from relaxing almost 100% in any age group. However, atriopeptin III-mediated responses were similar in the three age groups and were unaffected by methylene blue. These results suggest that 1) sensitivity to SNP increases with age from fetal through adult life; 2) relaxation mediated by atriopeptin III is similar during development; 3) methylene blue does not affect SNP mediated relaxation in fetuses but progressively decreases sensitivity to SNP in newborns and adults; and 4) methylene blue does not affect atriopeptin III-mediated relaxation in any age group.