James Humme

Effect of fetal adrenalectomy on catecholamine release and physiologic adaptation at birth in sheep.

Plasma catecholamine levels increase dramatically at birth. To determine the contribution of adrenal catecholamine secretion to the surge in catecholamines at birth and the role in newborn adaptation, we performed surgical adrenalectomy or sham operation on near-term ovine fetuses. After recovery in utero, the animals were delivered and supported by mechanical ventilation. Plasma catecholamine levels, heart rate, blood pressure, cardiac output, pulmonary function, surfactant secretion, and release of free fatty acids (FFA) and glucose were compared in control and adrenalectomized animals. Plasma epinephrine increased rapidly at birth in controls but was undetectable in adrenalectomized animals. Norepinephrine levels were not statistically different. Heart rate, blood pressure, cardiac output and contractility increased abruptly after cord cutting in controls but did not increase in adrenalectomized animals. Lung compliance, pulmonary function, surfactant pool size, glucose and FFA levels were significantly decreased in adrenalectomized animals. These results suggest that adrenal epinephrine secretion is vital to many of the adaptive events at birth.

The effects of hypoxia on (methionine) enkephalin peptide and catecholamine release in fetal sheep

ABSTRACT: The effect of hypoxia on plasma met-enkephalin and catecholamine levels was studied in chronically catheterized fetal sheep. Maternal and fetal hypoxia was maintained for 20 min. We found hypoxia significantly increased the plasma levels of large mol wt met-enkephalin containing peptides from 1755 ± 229 pg/mL during baseline to 4408 ± 1426 pg/mL by 15 minutes of hypoxia. The levels of the met-enkephalin pentapeptide were unchanged during hypoxia from a baseline value of 168 ± 56 pg/mL. Norepinephrine and epinephrine levels increased 5- and 10-fold, respectively, by 15 min of hypoxia. These observations suggest cosecretion of the large mol wt met-enkephalin peptides with catecholamines during stress in developing animals.

The Effect of Chemical Sympathectomy on Catecholamine Release at Birth

ABSTRACT. The precise source of circulating catecholamine (CA) at birth and their role in circulatory adaptation is unclear. In order to determine the contribution of increased postganglionic sympathetic nerve activity to the CA surge at birth, we induced complete sympathectomy in near term fetal lambs prior to delivery by giving 6-hydroxydopamine. Chronically catheterized fetal sheep received either 6-hydroxydopamine (n=5) or control infusion (n=6). Chemical sympathectomy was verified by tyramine infusion. Lambs were delivered at 142 ± 1 days of gestation and serial plasma CA, heart rate, blood pressure, cardiac output, blood gases, blood glucose, and free fatty acids, were measured before and for 4 h after delivery. Myocardial ß-adrenergic receptors and tissue CA concentration were determined following sacrifice. Baseline circulating norepinephrine (NE) values were lower in sympathectomized animals (183 ± 45 versus 373 ± 125 pg/ml, p<0.05) and epinephrine values were slightly higher (118 ± 89 versus 48 ± 1 pg/ml, NS). There was only a 2-fold increase in NE after cord cutting in sympathectomized animals while control animals had a 4-fold increase (peak NE values 354 ±121 versus 1305 ± 363 pg/ml respectively, p<0.001). Epinephrine increased significantly in both groups and there were no significant differences between sympathectomized and control animals. Heart rate and blood pressure rose abruptly in both groups after cord cutting and there were no significant differences. Cardiac output was similar in both groups. Blood glucose in sympathectomized animals was significantly higher than control before (p<0.005) and after (p<0.001, peak values 66 ± 15 versus 35 ± 12 mg/dl). Plasma free fatty acid levels rose after cord cutting and were similar in both groups. Tissue NE content in the hearts of sympathectomized animals were less than 5% of control (mean 1.92 ± 0.5%), verifying the completeness of sympathectomy in this tissue. Myocardial ß-adrenergic receptors were 229 ± 21 versus 199 ± 26 fmol/mg protein in sympathectomized and control animals, respectively. These results suggest that the majority of circulating NE seen at birth arises from postganglionic sympathetic neurons rather than adrenal medullary secretion. The comparable hemodynamic and metabolic homeostasis of the two groups underscores the importance of increased circulating adrenal epinephrine at birth.

The Contribution of Transporter-Dependent Uptake to Fetal Catecholamine Clearance

These studies were designed to determine the contribution of cocaine-sensitive, transporter-dependent, reuptake mechanisms to the intrauterine norepinephrine clearance rate in chronically catheterized fetal sheep. Baseline norepinephrine clearance and appearance rates were 125 +/- 20 ml/kg/min and 85 +/- 11 ng/kg/min, respectively. Transporter-dependent clearance represented 40% of the intrauterine clearance rate. The effects of chronic cocaine administration on fetal catecholamine clearance and appearance rate were then determined in animals treated with daily infusion of saline or cocaine. The total intrauterine clearance rate and the transport-dependent component of intrauterine clearance decreased significantly following the week of drug or placebo treatment, p < 0.05. This was associated with a threefold increase in circulating catecholamine concentrations in both groups of animals, p < 0.05. These results demonstrate that intrauterine catecholamine clearance is highly dependent on transporter-dependent mechanisms. Chronic intrauterine stress, manifested by increased circulating norepinephrine, is associated with a significant decrease in norepinephrine clearance and may be important in the pathogenesis of the adverse effects of stress on the fetus and of drugs like cocaine, which block catecholamine reuptake.

Naloxone potentiates epinephrine release during hypoxia in fetal sheep: dose response and cardiovascular effects

ABSTRACT: The effect of opiate receptor blockade on the plasma catecholamine response to hypoxemia was studied in seven chronically catheterized fetal lambs in utero. All animals underwent treatment with hypoxia alone, naloxone infusion alone (2 mg/kg) and hypoxia with naloxone at four different dosages (0.1, 0.5, 1.0, and 2.0 mg/kg). Maternal and fetal hypoxia was maintained for 20 min. There were no differences noted in the degree of hypoxemia or acidemia between the different hypoxia treatment groups. Hypoxia increased both norepinephrine and epinephrine plasma levels in all fetal sheep studied. We found a dose-dependent increase in plasma epinephrine levels in response to naloxone infusion during hypoxia. Plasma epinephrine level by 20 min of hypoxia with the 0.1 mg/kg naloxone dose (geometric mean 5366 pg/ml) was significantly more than with hypoxia alone (997 pg/ml). Naloxone at the other doses did not alter the epinephrine responses. There was no augmentation of plasma norepinephrine levels by naloxone at any dose studied. Thus, naloxone augmented the plasma epinephrine response to hypoxia in fetal sheep suggesting that the opiate peptides act as modulators of the sympathoadrenal system. The naloxone dose response differences observed in this study suggest this modulation is largely by antagonism of μ-receptors.

Metabolic Clearance and Plasma Appearance Rates of Catecholamines in Preterm and Term Fetal Sheep

Abstract: Plasma catecholamines increase markedly during labor and delivery. Moreover we have noted greater increases at birth in preterm than term lambs. It was unclear whether these differences were due to differences in secretion or clearance. We therefore designed experiments to compare the metabolic clearance rates (MCR) and plasma appearance rates of norepinephrine (NE) and epinephrine (E) in chronically catheterized term (143 ± 1 days) and preterm (131 ± 1 days) fetal sheep. Two different techniques, radioisotope tracer techniques and infusion of cold hormones for estimation of clearance rates, were compared systematically in the same group of animals. The results demonstrate that MCR of NE in term fetuses (178 ± 28 ml·kg−1 min−1) is similar to preterm fetuses (205 ± 22 ml·kg−1 min−1) as is MCR for E (193 ± 28 versus 170 ± 33 ml·kg−1 min−1, respectively). Estimates of MCR from cold hormone infusion were highly dependent on infusion rate with estimtes as much as 150% above that determined by isotope tracer infusions. Plasma appearance rates for both NE and E in term and preterm animals were similar. There were no detectable physiologic alterations during the tracer isotope infusions whereas infusions of cold hormone were associated with marked alterations in heart rate and blood pressure. These results suggest: 1) there are no differences in MCR or plasma appearance rates of NE or E between 130 days gestational age and term fetal sheep; 2) cold hormone infusions overestimate MCR and plasma appearance rates of both NE and E when compared to radioisotope tracer infusions; 3) differences in catecholamine concentrations at birth between term and preterm animals appear to be due to differences in secretion rather than clearance.

Placental norepinephrine transporter development in the ovine fetus

The placenta has been shown to be a site of expression of several of the monoamine membrane uptake transporters. However, the development and relative contribution of transport-dependent mechanisms to placental catecholamine clearance in vivo have not been demonstrated. These studies were designed to determine the development of the placental norepinephrine transporter (NET) and the relative contribution of transport dependent mechanisms to whole body and placental catecholamine clearance. Norepinephrine clearance and production rate were determined in 122 +/- 1 day gestation chronically catheterized fetal sheep. Placental clearance was shown to account for over 40 per cent of total intrauterine clearance and, of the clearance in the placenta, nearly 50 per cent was uptake, transport-dependent as shown by specific pharmacologic blockade. NET transport expression was examined by measurement nisoxetine binding in placenta and compared with binding in the frontal cortex of fetal, newborn and adult animals. Nisoxetine is a selective ligand for the norepinephrine transporter. Nisoxetine binding was 20-fold greater in placenta than in frontal cortex. Placental transporter binding decreased modestly in between 99 days gestation and term (145 days) but did not change in frontal cortex. These results suggest that expression of the norepinephrine transporter in the placenta is associated with a significant capacity for neurotransmitter re-uptake in utero. Given the high fetal norepinephrine production rate, this capacity is important for fetal homeostasis. This site of transporter expression may be important in the pathogenesis of derangements in catecholamine production in the fetus and in the adverse effects on the fetus of drugs, such as cocaine, which block catecholamine re-uptake.

Fetal plasma and renal responses to ruminal fluid.

Amniotic fluid homeostasis is dependent on a balance of fetal fluid production and absorption. The fetal gastrointestinal tract is believed to resorb 500 to 1000 ml of amniotic fluid per day during 7 to 10 bouts of swallowing activity. However, the impact of ruminal fluid on fetal plasma composition and fluid homeostasis is largely unknown. Seven ovine fetuses (120 +/- 1 day) received intraruminal infusions of 0.9% or 3% saline solution on alternate days. In response to successive 40-minute intraruminal infusions of 0.9% saline solution (0.5 and 1.0 ml/kg/min), there was no change from basal levels of fetal plasma osmolality (295.7 +/- 2.9 mosm), plasma arginine vasopressin (1.45 +/- 0.29 pg/ml), urine osmolality (150 +/- 8 mosm), or urine volume (0.49 +/- 0.10 ml/min). In response to the 3% saline solution infusion, significant increases were noted in fetal plasma osmolality (295.4 +/- 3.1 to 302.6 +/- 2.6 mosm), plasma arginine vasopressin (1.77 +/- 0.31 to 4.84 +/- 0.79 pg/ml), and urine osmolality (157 +/- 13 to 342 +/- 25 mosm), whereas fetal urine volume significantly decreased (0.35 +/- 0.05 to 0.15 +/- 0.06 ml/min). These results indicate that hypertonic, but not isotonic, saline solution infusion into the fetal gastrointestinal tract may affect fetal plasma composition and urine production. Under conditions of significant plasma to luminal osmotic gradients, fetal gastrointestinal water and electrolyte transfer may be more rapid than can be compensated by either fetal renal function or placental equilibration.