1,2-dibromo-3-chloropropane (DBCP)-induced infertility in male rats mediated by a post-testicular effect

The potential for the chemosterilant 1,2-dibromo-3-chloropropane (DBCP) to reduce male fertility by acting at a site in the genital tract beyond the testis was evaluated in male, Fischer 344 rats. A single sc treatment with 100 mg/kg DBCP reduced fertility in male rats 2 to 7 days postexposure without affecting mating frequency. Doses of 10, 20, or 40 mg/kg DBCP given sc once daily for 7 days caused a dose-dependent reduction in the metabolism of glucose to CO2 by epididymal sperm, as measured in vitro. Conversion of glucose to lactate was not reduced, indicating inhibition of energy metabolism at a step post-glycolysis. No clinical signs of toxicity were observed in these studies. Direct addition of DBCP to epididymal sperm being incubated in vitro also inhibited the metabolism of glucose to CO2. Inhibition was concentration related, and the minimal inhibitory concentration was 0.316 mM. These data indicate that DBCP may cause a nearly immediate infertility via a direct effect on post-testicular sperm. A possible mechanism of this infertility is inhibition by DBCP of glucose metabolism in the ejaculated sperm.     

Sites of inhibition of mitochondrial electron transport by rhein

The effect of rhein on the oxygen consumption, oxidative phosphorylation, ATPase activity and redox state of electron carriers of rat liver mitochondria has been studied. Rhein inhibits ADP- and uncoupler-stimulated respiration on various NAD-linked substrates and succinate, but stimulates state 4 respiration of mitochondria respiring on succinate. Experiments on specific segments of the respiratory chain showed that rhein does not inhibit electron flow through cytochrome oxidase. Electron flow through site 2, the ubiquinone-cytochrome b-cytochrome c1 complex, was also unaffected by rhein, which failed to inhibit the oxidation of duroquinol. Rhein affects oxidative phosphorylation by inhibiting both electron transfer and ADP-driven H+ uptake. The inhibition of succinate oxidation by rhein was found to take place at a point between succinate and ubiquinone, perhaps at the level of succinic dehydrogenase. Spectroscopic evidence demonstrated that rhein induces a NAD(P)H oxidation in mitochondria respiring either on endogenous substrates or on glutamate + malate, and an inhibition of the cytochrome b reduction by succinate. These observations, together with other evidence, suggest that rhein inhibits electron transport in rat liver mitochondria at the dehydrogenase-coenzyme level, particularly when the electron carriers are in a relatively oxidized state and/or when the inner membrane-matrix compartment is in the condensed state.     

Toxicologic and reproductive effects of inhaled 1,2-dibromo-3-chloropropane in male rabbits

Groups of 10 male New Zealand white rabbits were exposed by inhalation to 0, 0.1, 1.0 or 10 ppm of 1,2-dibromo-3-chloropropane (DBCP) vapor for 6 hours/day, 5 days/week for 14 weeks, except that the 10 ppm group was exposed for only 8 weeks due to mortality. The semen of rabbits was evaluated on a weekly basis during the exposure period and at periodic intervals during a recovery period (32 weeks for all groups except the 10 ppm groups which was for 38 weeks). In order to assess the fertility of the exposed rabbits, each male was allowed to mate with an unexposed female at the 14th and 41st week of the study. Exposure of rabbits to 1 and 10 ppm of DBCP by inhalation produced adverse reproductive effects as well as decreases in sperm count, motility and viability. Rabbits treated at 1 and 10 ppm had decreased sperm counts between the 8th and 14th weeks of the study. All of the 10 ppm rabbits were infertile when mated during the 14th week. The effects of DBCP on spermatogenesis were shown to be essentially reversible in rabbits exposed to 1 ppm; however, at 10 ppm, recovery was not complete under the conditions of the test. Rabbits exposed to 10 ppm had severe testicular alterations as early as 4 weeks into the study and these progressed to severe testicular atrophy by 8 weeks. Those exposed to 1 ppm for 14 weeks developed moderate testicular atrophy (approximately 50% reduction in size). Following the recovery period, the rabbits in the 10 ppm group had evidence of partial reversibility of the testicular atrophy. Electron microscopic evaluation of testicular tissue confirmed findings by light microscopy effects and also indicated increased numbers of abnormal sperm within the seminiferous tubules of rabbits at both the 10 and 1 ppm exposure levels. Those exposed to 0.1 ppm had an equivocal increase in abnormal sperm after the 14-week exposure period but not after the recovery period. Based on these results 0.1 ppm level of DBCP is considered as a no effect level for reproductive parameters.     

Tumor cell death induced by the inhibition of mitochondrial electron transport: The effect of 3-hydroxybakuchiol

Changes in mitochondrial ATP synthesis can affect the function of tumor cells due to the dependence of the first step of glycolysis on mitochondrial ATP. The oxidative phosphorylation (OXPHOS) system is responsible for the synthesis of approximately 90% of the ATP in normal cells and up to 50% in most glycolytic cancers; therefore, inhibition of the electron transport chain (ETC) emerges as an attractive therapeutic target. We studied the effect of a lipophilic isoprenylated catechol, 3-hydroxybakuchiol (3-OHbk), a putative ETC inhibitor isolated from Psoralea glandulosa. 3-OHbk exerted cytotoxic and anti-proliferative effects on the TA3/Ha mouse mammary adenocarcinoma cell line and induced a decrease in the mitochondrial transmembrane potential, the activation of caspase-3, the opening of the mitochondrial permeability transport pore (MPTP) and nuclear DNA fragmentation. Additionally, 3-OHbk inhibited oxygen consumption, an effect that was completely reversed by succinate (an electron donor for Complex II) and duroquinol (electron donor for Complex III), suggesting that 3-OHbk disrupted the electron flow at the level of Complex I. The inhibition of OXPHOS did not increase the level of reactive oxygen species (ROS) but caused a large decrease in the intracellular ATP level. ETC inhibitors have been shown to induce cell death through necrosis and apoptosis by increasing ROS generation. Nevertheless, we demonstrated that 3-OHbk inhibited the ETC and induced apoptosis through an interaction with Complex I. By delivering electrons directly to Complex III with duroquinol, cell death was almost completely abrogated. These results suggest that 3-OHbk has antitumor activity resulting from interactions with the ETC, a system that is already deficient in cancer cells.     

Atropine-induced inhibition of sperm and semen transport impairs fertility in male rats

Previous studies revealed that atropine reduced male fertility in rats without any effects on mating performance, sperm production and motility, and testicular morphology. The present study was conducted to investigate whether the impairment of male fertility induced by atropine was related to the inhibition of sperm and semen transports from the vas deferens and seminal vesicle to the urethra during the process of emission. Male rats were treated with atropine at 125 mg/kg/day for 10-17 days prior to mating with untreated females. After confirmation of mating, male rats were euthanized and sperm number in the vas deferens and weights of the seminal vesicle and copulatory plug were determined as indicators of inhibition of sperm and semen transports, respectively. Reproductive status of mated females was determined on gestation days 15-17. A low pregnancy rate associated with a decreased number of implants was observed in females that mated with the atropine-treated males. The average number of sperm in the vas deferens was increased in the atropine-treated males. The average seminal vesicle weight in the atropine-treated males was greater than that of controls. The copulatory plug weights were decreased in the atropine-treated males. These results suggest that inhibitions of sperm and semen transports from the vas deferens and seminal vesicle to the urethra during the process of emission result in reduced male fertility in rats.     

The integrity of liver protein synthesis in male rats treated with 1,2-dibromo-3-chloropropane

Male rats treated with a single dose of 1,2-dibromo-3-chloropropane (DBCP) were tested for their ability to carry out the synthesis of liver proteins. In animals treated for 12 h, we found no changes in the uptake of [14C]orotic acid into liver RNA or the uptake of [3H]leucine into liver or serum protein. Uptake of [3H]leucine into the soluble fraction of the enlarged liver increased in proportion to liver size, while the uptake of [14C]orotic acid was unchanged. Examination of the ultrastructure of liver cells from rats treated for 12, 24, or 48 h revealed that the structure of the rough and smooth endoplasmic reticulum (RER; SER) were modified. An absence of ordered stacks of the RER and the presence of tangled nets of SER were noted.     

Acute nephrotoxicities and hepatotoxicities of 1,2-dibromo-3-chloropropane and 1,2-dibromoethane in male and female F344 rats

Four consecutive intraperitoneal (i.p.) injections with 40 mg/kg of 1,2-dibromo-3-chloropropane (DBCP) reduced the in vitro accumulation of p-aminohippurate (PAH) and tetraethylammonium (TEA) by slices of renal cortex and increased blood urea nitrogen (BUN) concentration in both male and female rats, but elevated serum glutamic pyruvic transaminase (GPT) and glutamic oxaloacetic transaminase (GOT) activities in females only. Four consecutive treatments with 1,2-dibromoethane (EDB) reduced the accumulation of PAH in male rats, but failed to alter TEA accumulation, BUN concentration or GPT and GOT activities in rats of either sex. Single i.p. injections of EDB or DBCP (40 mg/kg, approximately one-half of the acute, i.p. LD50 values) were without effect on serum GPT and GOT activities, BUN concentration or the accumulations of PAH and TEA in male rats when measured 24, 48 or 96 h after treatment, except that PAH accumulation was reduced at 96 h.These results indicate that BUN and the accumulations in vitro of PAH and TEA by renal cortical slices are appropriate endpoints for studying DBCP nephrotoxicity. Measurements of serum GOT and GPT activities detected DBCP hepatotoxicity in female rats only. The nephrotoxicity of EDB was indicated by measurement of TEA accumulation only.     

卵胞浆内单精子显微注射技术治疗男性不育127个周期的研究

目的常规体外受精与胚胎移植辅助生殖技术在治疗男性少、弱、畸精症和无精症时,常因受精障碍而失败。本研究对采用卵胞浆内单精子显微注射技术治疗男性不育的临床效果进行分析。方法采用常规超排卵方案,在B超引导下经阴道取卵,取新鲜精液或附睾睾丸取精,对MⅡ期成熟卵进行单精子显微注射。结果127个周期取1142个MⅡ期成熟卵进行单精子显微注射,受精率为75％,卵裂率为96％,临床妊娠率33％。结论卵胞浆内单精子显微注射技术是治疗男性不育的有效方法。     

Hydrogen sulfide-mediated stimulation of mitochondrial electron transport involves inhibition of the mitochondrial phosphodiesterase 2A,elevation of cAMP and activation of protein kinase A

Although hydrogen sulfide (H₂S) is generally known as a mitochondrial poison, recent studies show that lower concentrations of H₂S play a physiological role in the stimulation of mitochondrial electron transport and cellular bioenergetics. This effect involves electron donation at Complex II. Other lines of recent studies demonstrated that one of the biological actions of H₂S involves inhibition of cAMP and cGMP phosphodiesterases (PDEs). Given the emerging functional role of the mitochondrial isoform of cAMP PDE (PDE2A) in the regulation of mitochondrial function the current study investigated whether cAMP-dependent mechanisms participate in the stimulatory effect of NaHS on mitochondrial function. In isolated rat liver mitochondria, partial digestion studies localized PDE2A into the mitochondrial matrix. NaHS exerted a concentration-dependent inhibitory effect on recombinant PDE2A enzyme in vitro. Moreover, NaHS induced an elevation of cAMP levels when added to isolated mitochondria and stimulated the mitochondrial electron transport. The latter effect was inhibited by Rp-cAMP, an inhibitor of the cAMP-dependent protein kinase (PKA). The current findings suggest that the direct electron donating effect of NaHS is amplified by an intramitochondrial cAMP system, which may involve the inhibition of PDE2A and subsequent, cAMP-mediated stimulation of PKA.     

Biochemical evidence that high concentrations of the antidepressant amoxapine may cause inhibition of mitochondrial electron transport

Overdosage with the antidepressant amoxapine causes metabolic acidosis and may lead to brain damage and death. To better understand the metabolic disturbances caused by amoxapine overdose, its effects on three simple systems were studied: growth of Saccharomyces cerevisiae, mitochondrial energy metabolism, and an electron transport system in microsomal membranes. Growth of yeast on all substrates except lactate was inhibited by amoxapine at 50-100 micrograms ml-1. Growth on lactate was observed at 200 micrograms ml-1 of amoxapine. In beef heart mitochondria, amoxapine at 100 micrograms ml-1 inhibited reactions involving large sections of the electron transport chain. Energy-linked reactions in submitochondrial particles were also inhibited. Electron microscopy showed some disruption of the mitochondrial internal structure by amoxapine and a change from orthodox to condensed conformation. Microsomal NADH-cytochrome b5 reductase was inhibited by amoxapine, but at higher amoxapine concentrations than mitochondrial reactions. The results suggest amoxapine disrupts reactions of membrane-associated enzyme complexes, and mitochondrial energy conservation may be one of the first systems affected. We speculate that lactic acid accumulation in patients with amoxapine overdose may be caused by loss of electron acceptor activity in tissues.     

Respiratory pathology in rats and mice after inhalation of 1,2-dibromo-3-chloropropane or 1,2 dibromoethane for 13 weeks

Seventy F344 rats and 144 B6C3F1 mice were subdivided into seven groups. Three groups were each exposed via inhalation to 1, 5, or 25 ppm of 1,2-dibromo-3-chloropropane (DBCP) for 6 h per day, 5 days per week for 13 weeks. Three additional groups were each similarly exposed to 3, 15, or 75 ppm of 1,2-Dibromoethane (EDB). The remaining group was exposed to room air under the same conditions. At 13 weeks, rats and mice showed severe necrosis and atrophy of the olfactory epithelium in the nasal cavity after inhalation of 5 or 25 ppm DBCP and 75 ppm EDB. Lower concentrations induced squamous cell metaplasia, hyperplasia and cytomegaly of the epithelium of the respiratory nasal turbinals. Squamous metaplasia, hyperplasia and cytomegaly of the epithelium was also seen in larynx, trachea, bronchi and bronchioles. Other compound related toxic lesions in rats were seen in the liver, kidney and testes.     

Acute toxicity of 1,2-dibromo-3-chloropropane in the F344 male rat: I. Dose-response relationships and differences in routes of exposure

Single treatments of F344 male rats with the nematocide 1,2-dibromo-3-chloropropane (DBCP) produced acute injury to the kindney, epididymis, testis, and liver. Severity was proportional to the size of the dose administered, but the kidney was adversely affected at a lowere single dose (40 mg/kg) than was the testis or liver (80 mg/kg). Primary target cells in the kidney were proximal tubular epithelia in the outer medulla. Disturbances in renal function included impaired tubular reabsorption (e.g., glucose, fluid, and electrolyte transport) and reduced glomerular filtration. Mild, hepatocellular swelling was produced by 40 mg/kg DBCP, while the leakage of intracellular enzymes into the blood, and centrolobular hepatocellular necrosis occurred after 80 and 120 mg/kg, respectively. The acute testicular lesion was characterized by degeneration and desquamation of the caput epididymal (head of the epididymis) epithelium and by disruption of the seminiferous tubular architecture; these effects were more severe after 120 mg/kg than after 80 mg/kg. Comparisons of the acute toxicity of DBCP following repeated sc, po (by gavage), and ip administration (40 mg/kg daily for 4 days) failed to reveal qualitative differences attributable to the route of exposure. However, the severity of the renal lesion appeared comparatively greatest when treatment occurred sc. These results indicate that DBCP is a cytotoxicant for epididymal and renal proximal tubular epithelia and, since lesions were produced by repeated exposure to acutely subtoxic doses, suggest that its effects on these tissues may be cumulative.     

ATP depletion does not account for apoptosis induced by inhibition of mitochondrial electron transport chain in human dopaminergic cells

As the mitochondrial electron transport chain (ETC) is necessary for life, its inhibition results in cell death. To date, ETC complex (I-IV) inhibitors (ETCIs) have been thought to induce ATP depletion, triggering cellular apoptosis. To clarify whether the depletion of intracellular ATP is relevant to apoptosis induced by ETCIs, we conducted comparative studies using oxidative phosphorylation inhibitors (OPIs), including a specific F(0)F(1)ATP synthase inhibitor oligomycin, an ionophore valinomycin and an uncoupler 2,4-dinitrophenol, as tools to deplete only ATP without influencing the ETC. In human dopaminergic SH-SY5Y cells, ETCIs (rotenone, thenoyltrifluoroacetone, antimycin A and potassium cyanide) depleted ATP and induced apoptosis. However, OPIs failed to induce apoptosis despite ATP being decreased to an extent comparable to that observed with ETCIs. Reactive oxygen species (ROS) production was augmented by ETCIs, but not by OPIs. Furthermore, ETCI-induced apoptosis was inhibited by the addition of an antioxidant N-acetylcysteine. Apoptosis was induced without ATP depletion by H(2)O(2) at a concentration that generated ROS at an amount comparable to that induced by ETCIs. Our findings demonstrate that ROS production is more relevant than ATP depletion to apoptosis induced by ETCIs.     

Structure-activity relationships for N,N'-bis(dichloroacetyl) diamines and substituted naphthoquinones in the inhibition of mitochondrial electron transport

The inhibition of mitochondrial electron transport was measured with four homologous series of N,N′-bis(dichloroacetyl) diamines and one homologous series of substituted naphthoquinones. Partition coefficients between acetone-water and silicone for these compounds were estimated by their R_m values in reversed-phase thin-layer chromatography. Parabolic correlations between a log function of biological activity and R_m were obtained with all series. A statistical analysis suggested that a homologous series of secondary amide derivatives of the N,N′-bis(dichloroacetyl) diamines differed from 3 homologous series of tertiary amide derivatives of the N,N'-bis(dichloroacetyl) diamines. Although N,N'-bis(dichloroacetyl) diamines and substituted napthoquinones affected the respiratory chain at different points and the substituted naphthoquinones were more effective inhibitors of electron transport, R_m values for maximum inhibition differed by only two methylene elements between the most widely separated series. Similarities between R_m values for maximum inhibition suggest that penetration of the mitochondrial membrane is a highly significant determinant for structure-activity relationships.     

Resveratrol in cancer: cellular and mitochondrial consequences of proton transport inhibition

Transformed cells suffer several changes leading to the increase of protective mechanisms and show a metabolic profile in accordance with higher proliferative capacity. In these mechanisms, changes in mitochondrial activity cause a higher glycolytic metabolism in detriment of oxidative phosphorylation. In these changes, H?-ATPase regulation seems to be importantly involved. During the last years, polyphenols and specially the stilbene resveratrol and related members of its family have been studied because they are able to affect tumour cell growth and cancer progression. Among the different effects induced by resveratrol, inhibition of H?-ATPase seems to be one important mechanism in its effect on cancer progression. Further, an ectopic H?-ATPase located in the outer surface of plasma membrane has been recently involved in cancer progression and angiogenesis. In this article we review the latest findings about resveratrol inhibition of H?-ATPase and its importance in tumour cell growth and cancer progression.     

Effects of L-carnitine administration on mitochondrial electron transport activity present in human muscle during circulatory shock

Carnitine was administered to a group of patients in shock, and the activities of cytochrome oxidase and succinate cytochrome c reductase in muscle needle biopsies from these patients were compared to those activities present in a non-carnitine treated control group of patients. Carnitine seemingly exerted a significant protective action on cytochrome oxidase activity during the initial phases of shock, but not to such an extent on succinate cytochrome c reductase activities.     

睾丸细针抽吸取精行卵胞浆内单精子注射治疗严重男性因素不育

目的 了解睾丸细针抽吸取精(Fine needle aspiration，FNA)结合卵母细胞单精子显微注射技术(Intracytoplasmic sperm injection，ICSI)治疗严重男性因素不育的技术成效及其治疗效果。方法 实施FNA结合ICSI技术治疗20例21个周期严重男性因素不育的夫妇，采用长方案超排卵，对处于细胞分裂中期(MetaphaseⅡ，MⅡ)的成熟卵母细胞行单精子显微注射。结果 共注射了239个MⅡ卵母细胞，其中149个正常受精，正常受精率为62．30％，临床妊娠9例，已分娩9个婴儿。结论 FNA结合ICSI技术给严重男性不育症患者提供了一种快速、方便、无痛、有效的治疗方法。     

The comparative effects of 1,2-dibromo-3-chloropropane (DBCP) and its metabolites, 3-chloro-1,2-propaneoxide (epichlorohydrin), 3-chloro-1,2-propanediol (alphachlorohydrin), and oxalic acid, on the urogenital system of male rats

Reported similarities in the acute toxic effects of 1,2-dibromo-3-chloropropane (DBCP), 3-chloro-1,2-propaneoxide (epichlorohydrin, ECH), 3-chloro-1,2-propanediol (alphachlorohydrin, ACH), and oxalic acid (OA) have been suggested as presumptive evidence that the metabolism of DBCP to OA, via ECH and ACH, is the cause of the resulting injuries to the kidney and, perhaps, to the epididymis and testis. To test this hypothesis, the comparative toxicities of these four chemicals were studied in male rats after single subcutaneous (sc) injections of maximally tolerated (nonlethal) doses. Kidney, testicular, and liver functions were monitored, and the occurrences of morphological changes in these and several other organs were evaluated 24 hr, 3, 8, 25, and 75 days post-treatment. DBCP caused renal dysfunction (alterations in urine composition and reduced glomerular filtration rate) and marked necrosis of the proximal tubular epithelium in the outer medulla of the kidney. ACH and OA also elicited renal dysfunction, but ACH produced only a mild swelling of the proximal tubular epithelium in the renal cortex and OA produced a focal necrosis anatomically associated with crystal deposition. ECH caused a swelling of the proximal tubular epithelium in the renal cortex, but not frank kidney dysfunction. DBCP also caused a reversible vacuolization of the tubular epithelium in the caput epididymis, progressive testicular atrophy, and a reduction of cauda epididymal sperm concentration. ACH and ECH produced similar effects, as well as epididymal sperm granulomas, spermatocoeles, and an increase in the number of morphologically abnormal spermatozoa. OA failed to produce discernible epididymal or testicular lesions at any time during the study. The development of similar lesions in the epididymis and testis following DBCP, ECH, or ACH treatments is consistent with the theory of metabolism of these chemicals to a common causative gonadotoxic agent. Oxalic acid (OA), however, would not appear to be the common gonadal toxicant. Differences in the effects, both morphological and functional, of DBCP, ECH, ACH, and OA on the kidney, moreover, indicate that DBCP nephropathy is not mediated through metabolism to OA and suggest, as well, that it differs causally from that induced by ECH or ACH. Therefore, the metabolism of DBCP to ECH or ACH, and of ECH or ACH to OA, is insufficient to explain totally the toxic effects of these agents on the urogenital system in male rats.     

Investigating ROS sources in male infertility: A common end for numerous pathways

Reactive oxygen species (ROS) are active byproducts of aerobic metabolism. Although they are constantly produced during normal cellular activities in the mitochondria, their action is counteracted by inherent antioxidant systems. This equilibrium is distorted in the case of acute or chronic inflammation, which results in increased ROS production and, ultimately, oxidative stress. In sperm, ROS are produced by both spermatozoa and circulating leucocytes and may be part of normal adaptive reactions, such as the capacitation process. However, a number of external toxicants may also contribute to ROS production in the testis and epididymis, leading to a decrease in sperm viability and motility and, therefore, an increased onset of the male factor of infertility. Such pro-oxidative conditions include, among others, exposure to radiation, extreme temperature, certain drugs, toxins, heavy metals, smoking and biological hazards. The current review paper summarizes the available evidence incriminating ROS and oxidative stress as the underlying pathophysiological mechanism leading to the onset of reproductive toxicity in each of these settings.     

Inhibition of central nervous system aromatase activity: a mechanism for fenarimol-induced infertility in the male rat

Fenarimol (alpha-(2-chlorophenyl)-alpha(4-chlorophenyl)-5-pyrimidine-methanol), a pyrimidine carbinol agricultural fungicide, was previously reported to cause a dose-related decrease in fertility in rats (K. S. Hirsch, E. R. Adams, D. G. Hoffman, J. K. Markham, and N. V. Owen (1986), Toxicol. Appl. Pharmacol. 86, 391-399). Based on the results of a number of reproduction studies (K. S. Hirsch, E. R. Adams, D. G. Hoffman, J. K. Markham, and N. V. Owen (1986), Toxicol. Appl. Pharmacol. 86, 391-399), the infertility appeared to be associated with an impairment of male sexual behavior. When [14C]fenarimol was administered to the dam, high concentrations of radioactivity were observed in the neonatal hypothalamus, which functions in the development and subsequent expression of male sexual behavior. In the present studies fenarimol exhibited neither antiandrogenic nor antiestrogenic activities. The compound did, however, prevent the increase in nuclear estrogen receptors in the brain which normally occurs in the male during the early postnatal period. These results suggested that fenarimol might be acting to inhibit estrogen biosynthesis (via the aromatase enzyme complex) within the central nervous system. [3H]Testosterone was administered to neonatal rats, and the tritiated metabolites were isolated. Testosterone and dihydrotestosterone (17 beta-hydroxy-5 alpha-androstan-3-one) concentrations were similar in all treatment groups. Tritiated estrogens were detected in the brain cell nuclei from control neonates but not in neonates exposed to fenarimol. Fenarimol was also observed to inhibit rat ovarian aromatase activity in vitro. These data indicate that the decrease in male sexual behavior and the infertility associated with exposure to fenarimol were, most likely, due to inhibition of aromatase activity within the central nervous system.