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In ovo exposure quail assay for risk assessment of endocrine disrupting chemicals.

Although there are in vivo assays using various organisms for the risk assessment of chemicals with endocrine disrupting properties, effective experimental methods for avian species are still under debate. We have developed an in ovo exposure assay using Japanese quail eggs, aimed at assessing disrupting effects on avian reproductive development and function. Hybrid eggs from Brazilian Brown male and White Egg female quails, which can be genetically sexed by their plumage color after hatching, were prepared, and test materials dissolved in olive oil were injected into the air-chamber on day 10 of incubation. After sexual maturation of hatched chicks, we observed egg production by females and the egg quality and male-typical reproductive behavior, and then examined reproductive system morphology and serum steroid concentrations in both sexes. Treatment with a synthetic estrogen, diethylstilbestrol (DES, 0.5-50 ng/g egg), dose-dependently reduced the eggshell thickness and strength of eggs. A few females treated with 5 ng/g DES per egg produced soft-shelled/ unmarked eggs, and all laying females treated with 50 ng/g egg produced eggs completely lacking shells. DES also induced shortening of the left oviduct and abnormal development of the right oviduct in a dose-dependent manner, while testis weight was reduced symmetrically. In addition, 2,2',4',6'-tetrachlorobiphenyl-4-ol (10-1,000 ng/g egg), which previously showed relatively high estrogenic activity in vitro, caused dose-dependent shortening of the left oviduct and reduction in testis weight. The methods for evaluating endocrine disrupting effects and preparing experimental birds proposed in the present study are expected to facilitate assays for avian reproductive toxicology.

Animals↗

Estrogen and endogenous opioids regulate CCK in reproductive circuits.

This review focuses on the interaction of estrogen with the cholecystokinin (CCK) and endogenous opioid peptide systems in the medial preoptic nucleus, and how these interactions result in alterations of a stereotypic female reproductive behavior--lordosis. The medial preoptic nucleus is an integral part of a circuit controlling lordosis that extends from the limbic system through the hypothalamus. Estrogen alters the integration of sensory information in the circuit that results in the display of sexually receptive behavior. Estrogen determines the activity of CCK and endogenous opioid peptide systems through regulation of expression, release and interaction with specific receptors. Studies of each system individually have indicated that they are pivotal to the expression of lordosis. Recent studies demonstrate an estrogen-dependent interaction between endogenous opioid and CCK systems that control reproductive behavior.

Animals↗

[Effect of p-chlorophenylalanine on rat reproduction under different conditions of lighting (author's transl)].

Reproduction in female rats kept on artificial cycles of light/darkness, continuous lighting and continuous darkness, has been studied. Three weekly doses of p-chlorophenylalanine (p-CPA), 100 mg/kg, were given to half of the rats in each group during the two preconditioning months before copulation. An evaluation has been made of the insemination, fecundation, number of embryos, ovarian cycle and level of 5-HT in the hypothalamus. The administration of p-CPA had an inhibitory effect on the reproductive behavior, especially pronounced in rats kept under continuous lighting and under continuous darkness. The difference in 5-TH levels were greater among the groups kept on cycles of light/darkness without treatment and with p-CPA (0.05 greater than P greater than 0.01). The importance of 5-HT in the mechanisms of reproductive behavior is emphasized.

Animals↗

Preoptic projections to Barrington's nucleus and the pericoerulear region: architecture and terminal organization.

The medial preoptic area (MPO), a sexually dimorphic region, plays a pivotal role in neuroendocrine function and reproductive behavior. We recently reported that MPO projects heavily to the midbrain periaqueductal gray (PAG). We also noted that MPO projects to the dorsolateral pontine tegmentum. Here we identified the cells of origin of the MPO-->tegmental projection and delineated the terminal organization of MPO projections to Barrington's nucleus, locus coeruleus (LC), and the rostromedial pericoerulear region (pLCrm). Correlative cyto- and chemoarchitectonic studies were done to define better the nuclear groups of the dorsolateral pontine tegmentum. Retrograde tracing revealed that MPO neurons projecting to the dorsolateral pontine tegmentum are preferentially distributed in distinct subregions of MPO, including the sexually dimorphic medial preoptic nucleus (MPN). Anterograde tracing with wheat germ agglutinin-horseradish peroxidase or Phaseolus vulgaris leucoagglutinin demonstrated considerable target specificity in projections from MPO to the dorsolateral pontine tegmentum. Barrington's nucleus receives a dense focal input along its entire rostrocaudal axis. In addition, pLCrm is heavily targeted by MPO inputs; pLCrm contains a concentrated plexus of extranuclear dendrites of LC neurons. The lateral dorsal tegmental (LDT) nucleus and LC proper receive only sparse input from MPO. MPO projections to Barrington's nucleus could regulate micturition reflexes during reproductive behavior. The MPO-->pLCrm projection could influence noradrenergic LC neurons in relation to reproductive and/or gonadal steroid function. Given the strong established connections from olfactory structures to MPO, it is possible that the MPO-->LC pathway provides an anatomical substrate for olfactory modulation of arousal.

Animals↗

A temperate reef fish, Tautogolabrus adspersus, (Walbaum) as a potential model species for laboratory studies evaluating reproductive effects of chemical exposure.

In ecotoxicological testing, there are few studies that report on reproductive output (egg production) of marine or estuarine fish. Cunner (Tautogolabrus adspersus) were studied as a potential model species to evaluate the impact of pollutants with estrogenic activity on reproduction in estuarine fish populations. Cunner inhabit marine and estuarine areas where contaminant discharges are likely. Baseline values for cunner gonadosomatic index (GSI), hepatosomatic index (HSI), and plasma vitellogenin (VTG) were determined in a field reference site (April 1999-December 1999). Male and female GSI indicated that cunner spawning is synchronized. Female HSI and VTG increased prior to GSI. From our laboratory observations, cunner are suitable for conducting experiments with reproductive endpoints indicative of both exposure (vitellogenin levels) and effects (egg production). However, cunner are not sexually dimorphic and stripping ripe fish is the only method to distinguish sex. In preparation for laboratory exposure studies with cunner, we designed a laboratory experimental holding system to accommodate cunner's reproductive behavior, a vertical spawning run to the water surface. Cunner were successfully acclimated from overwintering torpor to spawning condition in the laboratory by gradually changing the environmental conditions of fish held at winter conditions (4 degrees C and 9:15-h light:dark photoperiod) to spawning condition (18 degrees C and 15:9-h light:dark photoperiod). Our results show that cunner successfully spawned daily in the laboratory. They produced fertile eggs in our experimental system designed to accommodate cunner's vertical spawning runs, demonstrating that male and female reproductive behavior was synchronized in the laboratory. Our observations indicate that cunner would be a useful model species for evaluating reproductive effects of environmental contaminants in laboratory studies.

Animals↗

Endocrine changes associated with spawning behavior in male kokanee salmon (Oncorhynchus nerka) and the effects of anosmia.

This study examines the relationship between social status, behavioral activity, and hormone levels through the spawning cycle in male kokanee salmon (Oncorhynchus nerka). Shortly after arrival at the spawning grounds, male kokanee undergo dramatic changes in morphological and physiological condition: there is an increase in body depth, a bright silvery appearance is replaced by an intense red coloration, and spermiation commences. Males become associated with nesting females which they defend against other males. The androgens, testosterone and 11-ketotestosterone, decline slowly over the spawning period from high levels observed in the silvery males; gonadotropin (GtH II) and 17 alpha, 20 beta-dihydroxy-4-pregnen-3-one increase as males approach final maturation. Androgen and GtH levels in males placed in pens with females over the spawning bed were higher than those in males penned without females. Among males competing for females, amounts of milt and plasma concentrations of GtH were higher in those males that were successful in gaining access to nesting females. A marked reduction in milt and plasma hormones following cauterization of the olfactory nares indicates that olfactory stimuli play a role in mediating endocrine and spermiation responses to the nesting females. Anosmic males were less vigorous and persistent in their courtship, suggesting that, either a female pheromone and/or high levels of hormone are necessary to maintain full reproductive behavior in male kokanee. The role of socially induced hormone changes in modulating reproductive behavior is discussed.

Animals↗

Use of chemical communication by the subterranean rodent Ctenomys talarum (tuco-tuco) during the breeding season.

Solitary subterranean rodents with a low frequency of direct contact between conspecifics are expected to use chemical communication to coordinate social and reproductive behavior. We examined whether reproductive tuco-tucos (Ctenomys talarum) were able to discriminate the reproductive condition, sex, and source population of conspecifics by means of chemical cues contained in urine, feces, soiled shavings, or anogenital secretions. During preference tests in which animals had direct contact with these chemical cues, tuco-tucos were able to determine the reproductive condition of opposite sex conspecifics independent of the source of odor. When only olfactory cues were available, both sexes discriminated reproductive condition of opposite sex individuals using urine. Females were also able to discriminate the reproductive condition of males using soiled shavings. Females spent more time investigating male odors than female odors; except in the case of feces, breeding males spent similar amounts of time investigating male and female odors. No preferences were detected for opposite sex urine from members of an animal's own versus another population. The role of chemical cues in territory defense and breeding performance by this highly territorial subterranean rodent is discussed.

Animal Communication↗

Premature delivery and subsequent reproduction.

To examine the relationship between premature delivery and subsequent reproductive behavior, we attempted to trace 100 mothers four years after the birth of a surviving preterm or term infant. Seventy mothers were successfully traced. Maternal age, ethnicity, gravidity, parity, religion, and socioeconomic status, and the sex, birthweight, gestational age, mode of delivery, and hospital days of the index infant were jointly related to three outcome measures: occurrence of subsequent pregnancy, and, among those who again became pregnant, pregnancy interval and number of subsequent children. Among women who had one or more subsequent births during the follow-up period, those whose index infants were of lower gestational age had fewer subsequent births (r = 0.602, df = 32, p less than 0.011). No other variables, including cesarean birth, were significantly related to later reproductive behavior. These findings indicate that the birth of a premature infant may have a significant effect of decreasing or delaying subsequent reproduction. Through confirmatory studies are needed, awareness of this relationship may help obstetric perinatologists and neonatologists sensitively discuss plans for contraception and further childbearing with women who have experienced the stress of the birth of a premature infant.

Black or African American↗

Effects of alpha-methyl-para-tyrosine on monoamine levels in the Japanese quail: sex differences and testosterone effects.

Experiments were performed to obtain more information on the regulation by steroids of catecholaminergic systems in the brain of Japanese quail. Dose-response and time-response experiments were first performed to determine optimal conditions for measuring turnover in the quail brain. The norepinephrine and dopamine turnover were then estimated in microdissected brain nuclei of birds that were either sexually mature or gonadectomized or gonadectomized and treated with testosterone. Two major facts that bear direct relationship with the control of masculine reproductive behavior were demonstrated. On one hand, the dopamine turnover in the medial preoptic nucleus, a sexually dimorphic brain structure which is critically implicated in the control of copulatory behavior was much higher in male than in female quail irrespective of the hormonal condition of the birds. On the other hand, norepinephrine concentrations appeared to be higher in several nuclei of the female brain by comparison with males. These sex differences might represent part of the causal factors that underlie the sex dimorphism in reproductive behavior in quail.

Analysis of Variance↗

Localization of galanin and its binding sites in the quail brain.

Avian galanin was first isolated from the chicken intestine, and subsequently we isolated the same peptide from the quail oviduct. Galanin is known to be a multifunctional peptide in the neuroendocrine system in mammals, while little information is available for the galanin action in the central nervous system in birds. In the present study, therefore, we examined the presence and the localization of galanin and its binding sites in the quail brain. The binding of [125I]iodo-avian galanin to brain tissues specifically inhibited as a function of the concentrations of unlabeled avian galanin. In all of the observed brain regions, the number of galanin-binding sites when compared on the basis of unit weight was maximal in the interbrain including the preoptic and hypothalamic regions and minimal in the cerebellum. The Scatchard plot analysis of the galanin binding to the interbrain revealed that the apparent equilibrium constant of dissociation (Kd) was 0.281 (95% confidence interval, 0.234-0.351) nM, suggesting the presence of a single class of high-affinity binding sites. Although galanin-like immunoreactivity was found in several restricted regions throughout the brain, the most intense immunoreaction was present in the diencephalic region. Abundant immunoreactive cell bodies and/or fibers were localized in several preoptic and hypothalamic nuclei, i.e., the nucleus preopticus medialis (POM), the nucleus paraventricularis (PVN), the nucleus periventricularis hypothalami (PHN) and the nucleus infundibuli (IN), that are known to be involved in the control of reproductive behaviors as well as the pituitary hormone secretion. A substantial number of immunoreactive cell bodies and/or fibers was further observed within the septal nuclei in the telencephalon and several mesencephalic nuclei including the nucleus intercollicularis (ICo) involved in the control of vocalizations. These results suggest that avian galanin may act as an important factor to modulate the some reproductive behaviors and/or the pituitary hormone secretion in the bird.

Amino Acid Sequence↗

The male red-sided garter snake (Thamnophis sirtalis parietalis): reproductive pattern and behavior.

Among the small group of species (e.g., some temperate zone turtles, snakes, and bats) that exhibit a dissociated reproductive pattern, the red-sided garter snake (Thamnophis sirtalis parietalis) is probably the most well studied. For these species, courtship and mating occur immediately upon emergence from winter dormancy; the gonads remain essentially inactive. Male red-sided garter snakes are a particularly informative animal model for examining the role of neuroendocrine factors associated with reproductive physiology and behavior because unlike species that exhibit an associated reproductive pattern, in which sex steroids initiate and control sexual behavior, reproductive behavior in the male garter snake appears to be independent of circulating sex hormone control. In fact, the only factor associated with the initiation of courtship behavior and mating in the male garter snake is an extended period of low temperature dormancy followed by exposure to warm temperatures. Yet the presence of sex steroid-concentrating neurons within the pathways regulating courtship and mating suggests that sex hormones may be involved in the activation of sexual behavior. Although circulating androgens are elevated upon emergence from hibernation, the initiation of courtship behavior and mating appears to be independent of direct androgen control. Thus steroid hormones may have indirect effects on mating behavior in animals that display "dissociated" reproductive behaviors.

Animal Husbandry↗

Circulating gonadal steroid hormones regulate estrogen receptor mRNA in the male rat forebrain.

In male rats, the conversion of testosterone to estrogen via aromatization is a critical step in a number of androgen-mediated functions, especially reproductive behavior. Within the central nervous system (CNS), locally formed estrogen binds to its cognate estrogen receptor protein. Little is known about what factors regulate the expression of estrogen receptors in the male rat CNS. This study examined whether circulating male gonadal steroid hormones have a role in the regulation of estrogen receptor mRNA in brain regions critical for the expression of male reproductive behavior. Male rats were gonadectomized or sham operated, and 3 days later were sacrificed. Their brains were fixed by perfusion, frozen, and sectioned. Tissue sections were hybridized to an 35S-labeled 850 base cDNA probe, complementary primarily to the steroid binding domain of the estrogen receptor mRNA. Following post-hybridization washes, slides were dipped in photographic emulsion and exposed for 2 weeks. Estrogen receptor mRNA-containing neurons were observed in all brain regions previously shown by steroid hormone autoradiography to concentrate estrogen. Gonadectomy did not alter the number of estrogen receptor mRNA-producing neurons, but did produce a two-fold increase in the relative amount of estrogen receptor mRNA per cell in the medial preoptic nucleus, periventricular preoptic area, and bed nucleus of the stria terminalis. This study shows that circulating gonadal steroids down-regulate steady state levels of estrogen receptor mRNA within specific brain regions, and thereby have the potential to regulate the sensitivity of particular target regions in the CNS to estrogen.

Animals↗

Sexual behaviors and reproductive health outcomes: associations with wife abuse in India.

CONTEXT: Wife abuse has been associated with a variety of health concerns. Associations between abuse and reproductive health in India are not well known. OBJECTIVE: To examine relationships between men's reports of wife abuse and reproductive health issues in northern India. DESIGN: Structured face-to-face interviews were conducted as part of the male reproductive health supplement of the PERFORM System of Indicators Survey, a systematic multistage survey conducted in 1995-1996. SETTING: The northern state of Uttar Pradesh, one of the least developed states in India. PARTICIPANTS: A total of 6632 married men aged 15 to 65 years who lived with their wives and completed all survey questions for the study variables reported here. MAIN MEASURES: Physically and sexually abusive behaviors toward wives, sexual activities outside marriage, sexually transmitted disease (STD) symptoms, contraception use, unplanned pregnancies, and sociodemographic characteristics. RESULTS: Fifty-four percent of men reported not abusing their wives, while 17% reported physically but not sexually abusing their wives, 22% reported sexual abuse without physical force, and 7% reported sexual abuse with physical force. Abuse was more common among men who had extramarital sex (for sexual abuse using force: odds ratio [OR], 6.22; 95% confidence interval [CI], 3.98-9.72). Similarly, men who had STD symptoms were more likely to abuse their wives (with current symptoms: OR, 2.43; 95% CI, 1.73-3.42). Unplanned pregnancies were significantly more common among wives of abusive men, especially sexually abusive men who used force (OR, 2.62; 95% CI, 1.91-3.60). CONCLUSIONS: Wife abuse appears to be fairly common in northern India. Our findings that abusive men were more likely to engage in extramarital sex and have STD symptoms suggest that these men may be acquiring STDs from their extramarital relationships, thereby placing their wives at risk for STD acquisition, sometimes via sexual abuse. These abusive sexual behaviors also may result in an elevated rate of unplanned pregnancies.

Adolescent↗

Central neural mechanisms of progesterone action: application to the respiratory system.

Around the turn of the century, it was recognized that women hyperventilate during the luteal phase of the menstrual cycle and during pregnancy. Although a causative role for the steroid hormone progesterone in this hyperventilation was suggested as early as the 1940s, there has been no clear indication as to the mechanism by which it produces its respiratory effects. In contrast, much mechanistic information has been obtained over the same period about a different effect of progesterone, i.e., the facilitation of reproductive behaviors. In this case, the bulk of the evidence supports the hypothesis that progesterone acts via a genomic mechanism with characteristics not unlike those predicted by classic models for steroid hormone action. We recently, therefore, undertook a series of experiments to test predictions of those same models with reference to the respiratory effects of progesterone. Here we highlight the results of those studies; as background to and precedent for our experiments, we briefly review previous work in which effects of progesterone on respiration and reproductive behaviors have been studied. Our results indicate that the respiratory response to progesterone is mediated at hypothalamic sites through an estrogen- (E2) dependent progesterone receptor- (PR) mediated mechanism requiring RNA and protein synthesis, i.e., gene expression. The E2 dependence of the respiratory response to progesterone is likely a consequence of the demonstrated induction of PR mRNA and PR in hypothalamic neurons by E2. In short, we found that neural mechanisms underlying the stimulation of respiration by progesterone were similar to those mediating its reproductive effects.

Animals↗