Lack of feminization of the cremaster nucleus by prenatal flutamide administration in the rat and pig.

PURPOSE: The sexually dimorphic cremaster nucleus contains motoneurons that project via the genitofemoral nerve and theoretically direct androgen dependent testicular descent. The effects of flutamide on descent and masculinization of the cremaster nucleus were studied in the rat and pig. MATERIALS AND METHODS: Flutamide was given to pregnant rats and pigs on days 16 to 22 and 65 to 113 of gestation, respectively. Tissues were perfused and examined at birth (pigs) or at age 30 days (rats). Spinal cords were removed, sectioned and immunohistochemically stained for serotonin (rats) or substance P (pigs) to demarcate the position of the cremaster nucleus and allow the determination of cremaster motoneuron number. RESULTS: After exposure to flutamide testes were undescended in 6 of 9 rats and 7 of 10 pigs. Cremaster motoneuron number per nucleus were 288 +/- 22 in control versus 250 +/- 27 in flutamide treated rats, and 165 +/- 28 in control versus 148 +/- 24 in flutamide treated pigs. The decrease in motoneuron number by flutamide was significant in both species (p < 0.02) but it did not approach the levels in female rats (93 +/- 11) and pigs (57 +/- 12). Cremaster motoneuron number did not correlate with testicular position. Porcine undescended testes were associated with a significant increase in mean gubernacular volume. CONCLUSIONS: Unlike other sexually dimorphic spinal cord nuclei masculinization of the cremaster nucleus appears to be largely androgen independent and it does not correlate with ipsilateral testicular descent. These data suggest that androgens do not mediate descent of the testes via the efferent limb of the genitofemoral nerve.

Effects of dopexamine on hemodynamics and oxygen consumption after beta blockade in lambs.

OBJECTIVES: Dopexamine is a synthetic catecholamine with predominantly beta 2 and dopaminergic adrenergic receptor activities. We investigated its effects on systemic and myocardial hemodynamics and oxygen consumption (VO2) in a newborn species and studied its predominant mechanism of action. DESIGN: Prospective dose-response study with each animal serving as it own control. SUBJECTS: Eight chronically instrumented, unanesthetized lambs, 9 to 11 days of age. INTERVENTIONS: After surgical instrumentation and recovery for 72 hrs, animals were infused with dopexamine at increasing doses (1, 10, and 100 micrograms/kg/min) for 5 mins each before and after beta 1 (metoprolol) and beta 1, beta 2 (propranolol) adrenergic receptor blockade. All studies were performed during normoxia. MEASUREMENTS AND MAIN RESULTS: Heart rate (HR) increased with increasing infusion rates of dopexamine and systemic arterial pressure and vascular resistance decreased. Cardiac index, left ventricular pressure development, and systemic VO2 were unchanged, as was the rate x pressure product. Left circumflex coronary artery blood flow and myocardial VO2 were unaltered. After beta 1-blockade, dopexamine produced an increase in HR and decreased systemic arterial pressure and vascular resistance. After beta 1-adrenergic receptor blockade, no change was noted in systemic or myocardial VO2, coronary blood flow, or rate x pressure product. After beta 1, beta 2-blockade with propranolol, increasing infusion rates of dopexamine resulted in decreases in systemic pressure and vascular resistance. CONCLUSIONS: Dopexamine produced significant cardiovascular effects mediated primarily by beta 2-adrenergic receptors, and also produced residual peripheral arterial vasodilation after combined beta 1- and beta 2-blockade. The latter finding suggests that dopaminergic receptor stimulation may partly mediate dopexamine's effects in newborn lambs.

Effects of alveolar hypoxia on the pulmonary circulation and lung mechanics after cromolyn sodium and U-60,257 in lambs.

Because alveolar hypoxia (HYP) triggers pulmonary mast cell degranulation with elaboration of vasoactive mediators such as leukotrienes, we investigated the effects of aerosolized cromolyn sodium (CS), a mast cell stabilizing agent, and U-60,257(U) (a leukotriene blocker) on the circulation, lung mechanics and thromboxane (TXB2) levels in 11 lambs during acute exposure to HYP. Studies were performed in awake, chronically instrumented animals, once after placebo (saline) and again after CS (100 mg; n = 5) or U (90 mg; n = 6). Pulmonary arterial pressure increased 42% during HYP after saline, and 32% and 19% after CS and U, respectively. Pulmonary vascular resistance did not change during HYP after CS or U. Systemic arterial pressure was unchanged after saline and CS but decreased after U; systemic vascular resistance dropped after both CS and U. No changes were seen in tidal volume, lung compliance or airway resistance during HYP after saline or either drug, but minute ventilation increased during HYP in all studies. TXB2 increased during HYP after saline in both studies and was not altered by CS. In contrast, after U, TXB2 decreased. Thus, U more effectively blunted the pulmonary vascular response to HYP than CS and resulted in mild systemic hypotension. The drop in TXB2 after U suggests leukotriene-induced thromboxane synthesis contributes to regulation of pulmonary, and possibly, systemic vasoactivity.

Changes in estrogen metabolism after chronic oral contraceptive administration in the rhesus monkey.

The metabolism and elimination of estradiol (E2) and ethynylestradiol (EE2) were examined in adult female rhesus monkeys treated with two different combinational oral contraceptive agents at 10x the human equivalent dose. Ethynerone/mestranol (20:1), anagestone/mestranol (10:1), or vehicle was administered by gavage over a 10-year period on a cycling schedule of 21 days of dosing followed by 7 days without. At the end of the 28-day cycle, six monkeys in each group were anesthetized and administered a 14C-E2/3H-EE2 dose iv. Serial blood samples collected before and up to 6 hr after dosing were analyzed for total radioactivity and the percentage of parent compound and each metabolite was determined by HPLC. Radioimmunoassay of the baseline samples revealed that the plasma concentration of endogenous E2 was lower in the ethynerone/mestranol-treated group as compared to the vehicle control group. The total radioactivity derived from 14C-E2 was more rapidly eliminated from the plasma of the ethynerone/mestranol group than the control group. In addition, the percentages of HPLC-resolved E2 and EE2 were less in the treated group while the percentages of estrone, estrone glucuronide, and EE2 3-sulfate were enhanced as compared to the control group. The anagestone/mestranol group exhibited the same trends as the ethynerone/mestranol group but the data were not generally statistically different from the control group. These data indicate that chronically administered progestin/estrogen oral contraceptive agents reduce endogenous plasma E2 concentrations at least in part by enhancing the biotransformation of E2 to rapidly eliminated metabolites.

Maternal, fetal, and neonatal elimination of ethanol in nonhuman primates.

Three cynomolgus and 6 rhesus monkeys (106-160 days gestational age) were administered ethanol (0.8-1.5 g/kg i.v.) over a 30-min period. Blood samples were drawn under anesthesia from catheters in the maternal femoral artery, fetal umbilical circulation and/or neonatal umbilical artery. Comparison of maternal/fetal blood sample pairs from 60 to 300 min after initiation of the ethanol infusion revealed a significant correlation of blood ethanol concentration (r = 0.968, p less than 0.001). Thus, during the elimination phase, maternal blood ethanol concentrations are predictive of fetal blood ethanol concentrations. In three studies where the fetus was delivered 2 h after the infusion of ethanol, the neonatal rates of elimination were observed to be approximately 1/4 those of the mother. These data indicate that during the last third of nonhuman primate pregnancy, the maternal component of ethanol elimination appears to be the predominant factor in the overall rate of fetal ethanol elimination.

Transplacental pharmacokinetics and metabolism of diethylstilbestrol and 17 beta-estradiol in the pregnant rhesus monkey.

Experiments were performed to describe and compare the transplacental pharmacokinetics of the teratogen and transplacental carcinogen, diethylstilbestrol [4,4'-dihydroxy-alpha,alpha'-diethyl-trans, cis-stilbene (DES)], and the endogenous estrogen, 17 beta-estradiol (E2). Timed mated pregnant rhesus monkeys (119-137 days gestation) were anesthetized, and catheters were implanted in the maternal femoral artery and the interplacental fetal artery and vein using an extraamniotic technique. Single doses of either radiolabeled DES or E2 were administered via the maternal radial vein. Maternal plasma levels of labeled compound decreased rapidly after dose administration. Fetal plasma levels of radioactivity derived from either DES or E2 increased rapidly and then plateaued higher than maternal levels 1--2 h after dose administration. High pressure liquid chromatography of maternal and fetal plasma samples revealed both parent and conjugated metabolites of DES and E2. The principal metabolite of DES (DES monglucuronide) was radiolabeled and given to either the mother or the fetus iv. There was no significant cross-over of this metabolite in either direction. It is concluded that DES crosses the primate placenta in an unconjugated form and that, based on total radioactivity, placental transfer is similar to that of E2. The extensive fetoplacental metabolism of DES appears to be responsible for the greater half-life of this agent and its metabolites in the fetal circulation compared with the maternal circulation.

The metabolism of diethylstilbestrol in the rhesus monkey and chimpanzee.

Radiolabeled diethylstilbestrol (DES) was administered to one pregnant and three normal female rhesus monkeys. One normal female chimpanzee was also included in the study. Regardless of the mode of presentation (oral versus intravenous), the urine was the principal route of excretion for each species. The urine contained no non-polar radioactivity, and Sephadex LH-20 (MeOH/EtOH-50:50) resolved the radioactivity into five fractions (A, B, C, D, E). Fractions A,B, C, and D were hydrolyzable with beta-glucuronidase, and the principal aglycones were identified with GC/MS as cis-trans DES and dienestrol. The fecal excretory products were extracted with dimethoxy methane/methanol (50:50) and the radioactivity partitioned between benzene and H2O. The polar radioactivity was resolved by LH-20 (MeOH/EtOH-50:50) into chromatographic fractions similar to the urinary conjugates. These fecal conjugates were, however, less sensitive to beta-glucuronidase hydrolysis. The primary non-polar fecal radioactivity was chromatographically similar to DES (LH-20 and HPLC) in both species, and in the rhesus monkey the principal products identified were cis-trans DES and dienestrol.