Endocrine Control Of Reproduction Research Articles

Progesterone initiates tendril formation in the oviducal gland during egg encapsulation in cloudy catshark (Scyliorhinus torazame)

The diverse reproductive strategies of elasmobranchs (sharks, rays, and skates) have attracted research attention, but the endocrine control of reproduction is still incompletely known in elasmobranchs. By long-term monitoring of the egg-laying cycle in cloudy catsharks (Scyliorhinus torazame), we recently demonstrated a transient increase in plasma progesterone (P4) levels just prior to the appearance of the capsulated eggs in the oviducts. In the present study, we examined the in vivo effects of P4 administration in mature female cloudy catsharks. Although no capsulated eggs were observed following the implantation of P4-containing silicone tubing, we did find dark swollen oviducts in the abdominal cavity, in which clumps of long and coiled tendrils were observed. The tendril is an extension of the egg capsule, and the formation of the egg capsule begins with the tendril before main capsule formation. During the period of P4 implantation, the tendrils elongated, and their diameters were significantly increased on day 2 of treatment. Tendril formation was also confirmed on the day of endogenous P4 surge. Tendrils were not formed in catsharks implanted with estradiol-17β or testosterone. Histological analysis of the oviducal gland revealed that P4 implantation induced the secretion of tendril materials from the secretory tubules in the baffle zone, while the tendril materials were stored in the cytoplasm of the secretory cells under low P4 condition. Morphometrically, the area of secreted luminal materials in the secretory tubules was highly correlated to the diameter of tendrils. Our results suggest that the P4 surge during the egg-laying cycle serves as a trigger for egg capsule formation in the oviducal gland of cloudy catshark, but the hormonal signals were incomplete as the main capsule was not formed. Further studies are required to identify the hormones required for ovulation and formation of the main egg capsule.

Reptile assisted reproductive technologies: can ART help conserve 300 million years of evolution by preserving extant reptile biodiversity?

Biodiversity loss is the greatest environmental problem threatening ecosystem, animal, and human health. Anthropogenic induced changes to climate, habitat, disease, species distributions, poaching, and unsustainable trade have accelerated extinction rates in all vertebrates, including reptiles. Preventing reptile extinctions will require humans to acknowledge these losses and develop ex situ and in situ plans to preserve them. Assisted reproductive technologies (ART) are management tools used to protect numerous vertebrate taxa; however, progress in developing ART for reptiles has lagged. Creating functional and sustainable reptile ART will strengthen our conservation capacity by capturing genetic material from select individuals to overcome natural or manmade boundaries. Utilising short-term gamete storage and genome resource banking, in conjunction with timed artificial insemination (AI) or ex ovo incubation, could lead to profound advances in reptile conservation, mitigating the loss of reptile biodiversity. In this article, we review ART reptile research completed since the 1970s. Topics include AI, hormonal control of reproduction, gamete collection, gamete storage, and genome resource banking. Additionally, we review the potential application of advanced reproductive methodologies, including in vitro /ex ovo fertilisation, intracytoplasmic sperm injection, cloning (somatic cell nuclear transfer), and genetic editing.

Vitellogenesis in Blue Gourami is Accompanied by Brain Transcriptome Changes

The blue gourami (Trichogaster trichopterus) is a model for hormonal control of reproduction in Anabantidae fish, but also relevant to other vertebrates. We analyzed the female blue gourami brain transcriptome in two developmental stages: pre-vitellogenesis (PVTL) before yolk accumulation in the oocytes, and high vitellogenesis (HVTL) at the end of yolk accumulation in the oocytes. RNA sequencing of whole-brain transcriptome identified 34,368 unique transcripts, 23,710 of which could be annotated by homology with other species. We focused on the transcripts showing significant differences between the stages. Seventeen and fourteen annotated genes were found to be upregulated in PVTL and HVTL, respectively. Five nuclear transcripts, three of which contain the homeobox domain (ARX, DLX5, CERS6), were upregulated in PVTL. Additionally, several receptors previously known to be involved in reproduction were identified, and three of these, G-protein coupled receptor 54, Membrane progesterone receptor epsilon, and Gonadotropin-releasing hormone II receptor (GPCR, mPR, and GnRHR) were measured by quantitative RT-PCR in brain, pituitary, and ovary samples from PVTL and HVTL stage females. Of these, GPCR was highly expressed in the brain and pituitary as compared to the ovary in both PVTL and HVTL. GnRHR was highly expressed in the ovary compared to the brain and pituitary, and its levels in the brain were significantly higher in PVTL than HVTL. Brain mPR mRNA levels were likewise higher in PVTL than HVTL. In conclusion, this study details changes in the female blue gourami brain transcriptome through yolk accumulation in the oocytes and identifies key genes that may mediate this process.

SUN-478 The Role of Thimet Oligopeptidase (EP24.15) in Regulating Reproduction

For almost six decades, gonadotropin releasing hormone (GnRH) has been examined biochemically and physiologically in the hormonal control of reproduction. More recently, Kisspeptin (Kiss), neurokinin B (NKB) and dynorphin A (DynA) comprising KNDy neurons in the hypothalamus have been demonstrated to regulate gonadotropin releasing hormone (GnRH) release. To add to the complex circuitry of reproduction, and associated physiological processes, we sought to define the role of Phoenixin (Phx), a recently identified neuropeptide produced in the hypothalamus, that increases GnRH peptide, GnRH receptor and Kisspeptin mRNAs, increases GnRH secretion and mediates ovarian cyclicity. EP24.15 (EC 3.4.24.15, thimet oligopeptidase) is a neuropeptide processing metalloenzyme that is expressed throughout the hypothalamus and other brain regions. All neuropeptide hormones have their downstream signaling activity moderated enzymatically in the extracellular space though the specific enzymes remain unknown. EP24.15 is best known for regulating GnRH by degrading the hormone and thus, inactivating binding to the GnRH receptor. Likewise, we have proven that Kiss is likewise degraded by EP24.15 further pointing to a role in regulating reproduction. To validate both physiological and biochemical regulation of DynA and Phx, we determined if these neuropeptides and EP24.15 are co-expressed in the brain, (in addition to our previous studies of GnRH and Kiss) identifying a functional neuroendocrine circuit whereby EP24.15 could regulate reproduction. We have demonstrated Phx immunoreactivity in areas of the brain related to stress (amygdala and bed nucleus of the stria terminalis) and reproduction (hypothalamic arcuate nucleus). Biochemically, we used proteomics to identify new neuropeptide substrates, their binding to the active site and cleavage sites by high resolution 3D structural models, mass spectrometry, and enzyme kinetics. Two cleavage sites are present in Kiss and DynA whereas there is a single cleavage site in GnRH and Phx. Enzyme kinetic variables of DynA and Phx (Km and Vmax) are comparable to published values for GnRH and Kiss. The data demonstrates that EP24.15 cleaves DynA, and Phx in vivo as an additional layer to the elaborate mechanism of reproduction. Though NKB was co-expressed with EP24.1.5, it was not a substrate for the enzyme which allows for finer tuning of the reproductive axis. A better understanding of novel, regulated peptides that integrate stress and reproductive processes will provide insight into brain circuitry for moderating normal and aberrant reproductive function.

Role of Hormones in the Control of Reproductive Physiology and Reproductive Behavior

In multicellular organisms, intercellular mediators such as hormones or growth factors and morphogens play primary roles in development and reproduction. Evolution of the signaling pathways in which these mediators are involved has thus played an important role in the appearance and success of these species. There are many reproductive modes and behaviours in the animal kingdom. A reproductive behaviour can be defined as all the actions taken by an organism toward the generation of one or more other organism that possesses at least some of its genetic patrimony. From a strict evolutionary point of view, the goal of an organism is to spread its genetic patrimony as much as possible in the next generations of its kind. This egoistic behaviour typically benefits for the entire species since the individuals spreading the most their genetic patrimony is generally those with the best genes and are thus helping their species to be the most fitted possible. In the present review, I briefly describe the implication of hormones in the control of mammals Reproduction and in their Sexual Behavior. In addition, since most multicellular organisms exhibit sexual reproduction, I also take into account hormonal control of reproduction together with sexual behavior.

Endocrine Disruptor Compounds (EDC) Effects on Reproduction

A large number of industrial chemical products (paints, flame retardants, pharmaceuticals, pesticides) are suspected or proved to act as endocrine disruptor compounds (EDCs). In the present survey of the literature, the main issue is to consider how to classify individual products as EDC or not. In this perspective, the different mechanisms leading to endocrine disruption are described: either by direct interaction with hormone receptors leading to its stimulation or inhibition, or by effect on endogenous hormone concentration through stimulation or inhibition of its synthesis or its degradation, or its binding to transport binding proteins. The present review takes particularly in consideration the effects of EDCs on the endocrine control of Reproduction in human and animals. Indeed, a number of EDCs can act as anti-androgens, anti-estrogens, and steroidogenic enzyme inhibitors that interfere with steroid action or production and thus can alter reproductive health.

Perinatal programming of the orexinergic (hypocretinergic) system in hypothalamus and anterior pituitary by testosterone

Orexins A/B derived from hypothalamic prepro-orexin (PPO) are agonists for orexin receptors 1 (OX1) and 2 (OX2). Previously, we showed clear sex differences in the hypothalamic-pituitary-gonadal orexinergic system in adult rodents. Here, we studied the effect of sexual brain differentiation on the orexinergic system in neuroendocrine structures regulating reproduction. We evaluated: a: proestrous and neonatally androgenized female rats; b: adult males, untreated or gonadectomized in adulthood and injected with oil or estradiol and progesterone (E2/P4); c: control and demasculinized males (perinatally treated with flutamide and later castration) injected either with oil or E2/P4 in adulthood. Rats were sacrificed at 12:00 and 18:00h; blood samples and brains were collected. Hormones were measured using radioimmunoassay. PPO, OX1 and OX2 mRNAs were quantified by qPCR in medial basal hypothalamus, anterior hypothalamus, adenohypophysis, and cortex. Western blots for OX1 were done in the same structures. In normal females, gonadotropins surged at 18:00h coinciding with significant elevations of PPO, OX1 and OX2 mRNAs and OX1 protein in hypothalamus and pituitary; no increases were observed at noon. Afternoon changes were absent in masculinized females. Demasculinized males when treated with E2/P4 showed high PPO, OX1 and OX2 mRNAs and OX1 protein expression in hypothalamus and pituitary at 12:00 and 18:00h compared vehicle-treated controls. The same steroid treatment was ineffective in males with normal brain masculinization. Here we show that neonatal testosterone shapes the sexual differences in the hypothalamic-pituitary orexinergic system in synchronicity to establishing the brain sex differences of the reproductive axis. The female brain controls gonadotropin surges and concurrent elevations of all studied components of the orexinergic system, suggesting its participation as a possible link between food intake, behavior and hormonal control of reproduction.

Proteomic response of Macrobrachium rosenbergii hepatopancreas exposed to chlordecone: Identification of endocrine disruption biomarkers?

The present work is the first study investigating the impacts of chlordecone, an organochlorine insecticide, on the proteome of the decapod crustacean Macrobrachium rosenbergii, by gel-free proteomic analysis. The hepatopancreas protein expression variations were analysed in organisms exposed to three environmental relevant concentrations of chlordecone (i.e. 0.2, 2 and 20µg/L). Results revealed that 62 proteins were significantly up- or down-regulated in exposed prawns compared to controls. Most of these proteins are involved in important physiological processes such as ion transport, defense mechanisms and immune system, cytoskeleton dynamics, or protein synthesis and degradation. Moreover, it appears that 6% of the deregulated protein are involved in the endocrine system and in the hormonal control of reproduction or development processes of M. rosenbergii (e.g. vitellogenin, farnesoic acid o-methyltransferase). These results indicate that chlordecone is potentially an endocrine disruptor compound for decapods, as already observed in vertebrates. These protein modifications could lead to disruptions of M. rosenbergii growth and reproduction, and therefore of the fitness population on the long-term. Besides, these disrupted proteins could be suggested as biomarkers of exposure for endocrine disruptions in invertebrates. However, further investigations are needed to complete understanding of action mechanisms of chlordecone on proteome and endocrine system of crustaceans.

Pharmacoperones for Misfolded Gonadotropin Receptors.

The gonadotropin receptors (luteinising hormone receptor; LHR and follicle-stimulating hormone receptor; FSHR) are G protein-coupled receptors (GPCRs) that play an important role in the endocrine control of reproduction. Thus genetic mutations that cause impaired function of these receptors have been implicated in a number of reproductive disorders. Disease-causing genetic mutations in GPCRs frequently result in intracellular retention and degradation of the nascent protein through misfolding and subsequent recognition by cellular quality control machinery. The discovery and development of novel compounds termed pharmacological chaperones (pharmacoperones) that can stabilise misfolded receptors and restore trafficking and plasma membrane expression are therefore of great interest clinically, and promising in vitro data describing the pharmacoperone rescue of a number of intracellularly retained mutant GPCRs has provided a platform for taking these compounds into in vivo trials. Thienopyrimidine small molecule allosteric gonadotropin receptor agonists (Org 42599 and Org 41841) have been demonstrated to have pharmacoperone activity. These compounds can rescue cell surface expression and in many cases, hormone responsiveness, of a range of retained mutant gonadotropin receptors. Should gonadotropin receptor selectivity of these compounds be improved, they could offer therapeutic benefit to subsets of patients suffering from reproductive disorders attributed to defective gonadotropin receptor trafficking.

RXR/USP and EcR are critical for the regulation of reproduction and the control of JH biosynthesis in Diploptera punctata

During development and reproduction the response to ecdysteroids is mediated by a heterodimeric receptor complex comprising the retinoid X receptor/ultraspiracle (RXR/USP) and the ecdysone receptor (EcR). Here, the role of these receptors in the endocrine control of reproduction is examined in the cockroach Diploptera punctata. We report the sequence of four DpRXR and three DpEcR splice variants, including the first description of a Drosophila EcRB2-like isoform in a hemimetabolous insect. DpRXR and DpEcR are broadly expressed in the tissues of adult females, with relatively high transcript levels in the corpora allata (CA), nervous tissue and ovary. Developmental profiling revealed an inverse correlation between DpRXR and DpEcR expression and the activity of the CA. RNAi-mediated depletion of DpRXR and DpEcR did not affect oocyte growth, but inhibited oviposition and impaired chorion formation. Retained oocytes exhibited a degenerating follicular epithelium and were slowly resorbed. Treated animals showed significantly higher rates of JH biosynthesis and a decrease in ecdysteroid titers at the end of vitellogenesis. Reduction of DpRXR and DpEcR expression resulted in an upregulation of genes involved in JH production and a downregulation of allatostatin receptor mRNA in the CA. Treatment with dsRNA also affected the expression of genes downstream of JH in target tissues including vitellogenin and Krüppel-homolog 1 as well as Broad-Complex, an early ecdysone response gene. Overall, results suggest that DpRXR and DpEcR are not required early in the reproductive cycle when events are JH-dependent, but do mediate critical ecdysteroid feedback to the CA late in the gonadotropic cycle.

Melanin-concentrating hormone (MCH) and gonadotropin-releasing hormones (GnRH) in Atlantic cod, Gadus morhua: Tissue distributions, early ontogeny and effects of fasting

Melanin-concentrating hormone (MCH) is classically known for its role in regulating teleost fish skin color change for environmental adaptation. Recent evidence suggests that MCH also has appetite-stimulating properties. The gonadotropin-releasing hormone (GnRH) peptide family has dual roles in endocrine control of reproduction and energy status in fish. Atlantic cod (Gadus morhua) are a commercially important aquaculture species inhabiting the shores of Atlantic Canada. In this study, we examine MCH and GnRH transcript expression profiles during early development as well as in central and peripheral tissues and quantify juvenile Atlantic cod MCH and GnRH hypothalamic mRNA expressions following food deprivation. MCH and GnRH3 cDNAs are maternally deposited into cod eggs, while MCH has variable expression throughout early development. GnRH2 and GnRH3 mRNAs “turn-on” during mid-segmentation once the brain is fully developed. For both MCH and GnRH, highest expression appears during the exogenous feeding stages, perhaps supporting their functions as appetite regulators during early development. MCH and GnRH transcripts are found in brain regions related to appetite regulation (telencephalon/preoptic area, optic tectum/thalamus, hypothalamus), as well as the pituitary gland and the stomach, suggesting a peripheral function in food intake regulation. Atlantic cod MCH mRNA is upregulated during fasting, while GnRH2 and GnRH3 transcripts do not appear to be influenced by food deprivation. In conclusion, MCH might be involved in stimulating food intake in juvenile Atlantic cod, while GnRHs may play a more significant role in appetite regulation during early development.

Hormonal control of reproduction in small ruminants

Reproduction of small ruminants can be controlled by several methods developed in recent decades. Some of these involve administration of hormones that modify the physiological chain of events involved in the sexual cycle. Methods which utilise progesterone or its analogues are based on their effects in the luteal phase of the cycle, simulating the action of natural progesterone produced by the corpus luteum after ovulation, which is responsible for controlling LH secretion from the pituitary. Use of prostaglandins is an alternative method for controlling reproduction by eliminating the corpus luteum and inducing a subsequent follicular phase with ovulation. Finally, the discovery of the properties of melatonin in photoperiod-dependent breeding animals opened up a new methodology to control reproduction in these species, inducing changes in the perception of photoperiod and the annual pattern of reproduction. Use of hormones to induce oestrus has allowed increased use of artificial insemination in small ruminants, a very useful management tool, considering the difficulty of detecting oestrus in these species. At commercial level, synchronisation of oestrus allows control of lambing and kidding, with subsequent synchronisation of weaning of young animals for slaughter. Also, it allows more efficient use of labour and animal facilities. Multiple ovulation and embryo transfer programmes are also possible with the use of oestrus synchronisation and artificial insemination. Finally, hormonal treatments have also been used to induce puberty in ewe-lambs and doelings.

The endocrine control of reproduction in Nereidae: a new multi-hormonal model with implications for their functional role in a changing environment

Nereidae are vital to the functioning of estuarine ecosystems and are major components in the diets of over-wintering birds and commercial fish. They use environmental cues to synchronize reproduction. Photoperiod is the proximate cue, initiating vitellogenesis in a temperature-compensated process. The prevailing paradigm in Nereidae is of a single 'juvenile' hormone controlling growth and reproduction. However, a new multi-hormone model is presented here that integrates the environmental and endocrine control of reproduction. This is supported by evidence from in vitro bioassays. The juvenile hormone is shown to be heat stable and cross reactive between species. In addition, a second neuro-hormone, identified here as a gonadotrophic hormone, is shown to be present in mature females and is found to promote oocyte growth. Furthermore, dopamine and melatonin appear to switch off the juvenile hormone while serotonin and oxytocin promote oocyte growth. Global warming is likely to uncouple the phase relationship between temperature and photoperiod, with significant consequences for Nereidae that use photoperiod to cue reproduction during the winter in northern latitudes. Genotypic adaptation of the photoperiodic response may be possible, but significant impacts on fecundity, spawning success and recruitment are likely in response to short-term extreme events. Endocrine-disrupting chemicals may also impact on putative steroid hormone pathways in Nereidae with similar consequences. These impacts may have significant implications for the functional role of Nereidae and highlight the importance of comparative endocrinology studies in these and other invertebrates.

Comparative structure analyses of cystine knot-containing molecules with eight aminoacyl ring including glycoprotein hormones (GPH) alpha and beta subunits and GPH-related A2 (GPA2) and B5 (GPB5) molecules

BackgroundCystine-knot (cys-knot) structure is found in a rather large number of secreted proteins and glycoproteins belonging to the TGFbeta and glycoprotein hormone (GPH) superfamilies, many of which are involved in endocrine control of reproduction. In these molecules, the cys-knot is formed by a disulfide (SS) bridge penetrating a ring formed by 8, 9 or 10 amino-acid residues among which four are cysteine residues forming two SS bridges. The glycoprotein hormones Follicle-Stimulating Hormone (FSH), Luteinizing Hormone (LH), Thyroid-Stimulating Hormone (TSH) and Chorionic Gonadotropin (CG) are heterodimers consisting of non-covalently associated alpha and beta subunits that possess cys-knots with 8-amino-acyl (8aa) rings. In order to get better insight in the structural evolution of glycoprotein hormones, we examined the number and organization of SS bridges in the sequences of human 8-aa-ring cys-knot proteins having 7 (gremlins), 9 (cerberus, DAN), 10 (GPA2, GPB5, GPHα) and 12 (GPHβ) cysteine residues in their sequence.DiscussionThe comparison indicated that the common GPH-alpha subunit exhibits a SS bridge organization ressembling that of DAN and GPA2 but possesses a unique bridge linking an additional cysteine inside the ring to the most N-terminal cysteine residue. The specific GPHbeta subunits also exhibit a SS bridge organization close to that of DAN but it has two additional C-terminal cysteine residues which are involved in the formation of the "seat belt" fastened by a SS "buckle" that ensures the stability of the heterodimeric structure of GPHs. GPA2 and GPB5 exhibit no cys residue potentially involved in interchain SS bridge and GPB5 does not possess a sequence homologous to that of the seatbelt in GPH β-subunits. GPA2 and GPB5 are thus not expected to form a stable heterodimer at low concentration in circulation.SummaryThe 8-aa cys-knot proteins GPA2 and GPB5 are expected to form a heterodimer only at concentrations above 0.1 microM: this would be consistent with a short-term paracrine role but not with an endocrine role after dilution in circulation. Consequently, GPA2 and GPB5 could exert separate endocrine roles either during development and/or during adult life of both vertebrates and invertebrates.

Morphofunctional alterations in testicular cells of deslorelin‐treated boars: an immunohistochemical study

In this study we thoroughly scrutinized testes morphology and investigated whether treatment of recipient boars with gonadotropin-releasing hormone (GnRH)-agonist deslorelin could alter the expression of 3beta-hydroxysteroid dehydrogenase (3beta-HSD), luteinizing hormone receptors (LHRs), and androgen receptors (ARs) in testicular cells. An implant containing 4.7 mg of the GnRH-agonist deslorelin was subcutaneously inserted into crossbred male pigs at 91 and 147 days of age. Testicular traits, morphology of the testes, the proteins' expression, and testosterone concentration in blood plasma were analyzed in all boars after slaughter at 175 days of age. Histological analysis revealed significant alterations in both the interstitial tissue and seminiferous tubules of experimental animals after 28 and 84 days of deslorelin treatment. The intensity of the AR immunostaining within the testis appeared as a function of the severity of testicular dysgenesis. Time-dependent action of deslorelin on the expression of LHR and 3beta-HSD in Leydig cells was also detected. Staining for LHR and 3beta-HSD was very weak or the Leydig cells were immunonegative. Concomitantly, a significant decrease in plasma testosterone level was found in both groups of deslorelin-treated boars when compared with the control group. This is the first report showing the cellular distribution of AR, LHR, and 3beta-HSD in testicular cells of deslorelin-treated boars. It is concluded that morphological and immunohistochemical studies are important for the evaluation of testicular histoarchitecture and steroidogenic function. Subsequently, the endocrine control of reproduction in the GnRH-agonist deslorelin-treated males will be better understood.

Reproductive endocrinology of the largest dasyurids: Characterization of ovarian cycles by plasma and fecal steroid monitoring. Part I. The Tasmanian devil ( Sarcophilus harrisii)

There is a strong body of knowledge on the reproductive endocrinology of macropods, but little detailed information is available on the hormonal control of reproduction in other marsupials. This study used plasma and fecal sex steroid monitoring to characterize the estrous cycle of the largest extant dasyurid—the Tasmanian devil ( Sarcophilus harrisii). A pro-estrous pulse in plasma progesterone (1.33 ± 0.2 ng/ml) occurred several weeks prior to onset of the luteal phase (LP), resulting in a characteristic biphasic pattern during the estrous cycle. This brief, pro-estrous progesterone pulse was associated with a predominantly cornified vaginal smear, and copulation in females paired with males. Mean luteal progesterone concentrations (5.28 ± 0.8 ng/ml) were sustained and peaked around day 15 from luteal onset; thereafter, concentrations declined precipitously and returned to baseline around day 25. Females that did not produce young returned to estrus after 33.7 ± 5.9 days. Fecal 20α-OH-pregnanes analyzed in a pregnanediol assay (PgD) were excreted in consistently higher levels than 20-oxo-pregnanes, but the pattern was similar for the two metabolites, and significantly correlated with fluctuations in plasma progesterone. Fecal total estrogen concentrations were highest during the follicular phase (FP) and accompanied a pro-estrous pulse in fecal progestagens. The mean duration of the estrous cycle was ∼32 days, with a FP of around 14 days (range 8–23 days), and a luteal phase of around 18 days (range 12–25 days). There were no differences in the length of the LP between mated and non-mated cycles. Gestation length was 17.9 ± 1.0 days (range 14–22 days). Fecal steroid monitoring revealed significant differences between the pattern of progestagens and estrogen concentrations during the pregnant and non-mated estrous cycle, suggesting maternal endocrine recognition of pregnancy in the Tasmanian devil.

Effects of Mating on Plasma Concentrations of Testosterone, Cortisol, Oestrone Sulphate and 15‐Ketodihydro‐PGF2α in Stallions

Very little information is available regarding the physiological mechanisms involved in the normal sexual activity in the stallion and, in particular, the endocrine control of reproduction is still not clearly understood. This experiment was designed to determine the short-term effect of sexual stimulation on plasma concentrations of testosterone, cortisol, oestrone sulphate and 15-ketodihydro-PGF(2alpha) in stallions. Semen samples were collected from 10 lighthorse stallions of proven fertility using a Missouri model artificial vagina. At the same time, blood samples were collected from the jugular vein with heparinized tubes, 20 and 10 min before oestrous mare exposure, at exposure and 10, 20, 30 min after dismounting. Testosterone concentrations showed a sharp rise 10 min after mating (p < 0.001), reached a plateau, and then showed a further increase 30 min after mating (p < 0.001). Cortisol concentrations increased 10 min after mating (p < 0.001) and remained at high levels in the subsequent samples taken. A peak of oestrone sulphate was observed 10 min after mating (p < 0.001). 15-Ketodihydro-PGF(2alpha) concentrations decreased rapidly at the moment of the exposure of the stallions to an oestrous mare (p < 0.05), returned to pre-mating concentrations and then decreased again 30 min after mating (p < 0.05).

P.1.g.002 The orexin A plasma levels in patients with restrictive type of anorexia nervosa

Orexins A/B derived from hypothalamic prepro-orexin (PPO) are agonists for orexin receptors 1 (OX1) and 2 (OX2). Previously, we showed clear sex differences in the hypothalamic-pituitary-gonadal orexinergic system in adult rodents. Here, we studied the effect of sexual brain differentiation on the orexinergic system in neuroendocrine structures regulating reproduction. We evaluated: a: proestrous and neonatally androgenized female rats; b: adult males, untreated or gonadectomized in adulthood and injected with oil or estradiol and progesterone (E2/P4); c: control and demasculinized males (perinatally treated with flutamide and later castration) injected either with oil or E2/P4 in adulthood. Rats were sacrificed at 12:00 and 18:00 h; blood samples and brains were collected. Hormones were measured using radioimmunoassay. PPO, OX1 and OX2 mRNAs were quantified by qPCR in medial basal hypothalamus, anterior hypothalamus, adenohypophysis, and cortex. Western blots for OX1 were done in the same structures. In normal females, gonadotropins surged at 18:00 h coinciding with significant elevations of PPO, OX1 and OX2 mRNAs and OX1 protein in hypothalamus and pituitary; no increases were observed at noon. Afternoon changes were absent in masculinized females. Demasculinized males when treated with E2/P4 showed high PPO, OX1 and OX2 mRNAs and OX1 protein expression in hypothalamus and pituitary at 12:00 and 18:00 h compared vehicle-treated controls. The same steroid treatment was ineffective in males with normal brain masculinization. Here we show that neonatal testosterone shapes the sexual differences in the hypothalamic-pituitary orexinergic system in synchronicity to establishing the brain sex differences of the reproductive axis. The female brain controls gonadotropin surges and concurrent elevations of all studied components of the orexinergic system, suggesting its participation as a possible link between food intake, behavior and hormonal control of reproduction.

Organochlorine compounds in liver and concentrations of vitellogenin and 17β-estradiol in plasma of sea bass fed with a commercial or with a natural diet

Results from previous experiments directed to determine the effect of different nutritional factors or the effect of xenobiotics on hormonal control of reproduction, lead to the hypothesis that hormonal perturbations repeatedly observed in sea bass ( Dicentrarchus labrax) broodstock feeding commercial diets could have been caused by the presence of aryl hydrocarbon receptor (AhR) ligands, such as dioxins, furans and polychlorinated biphenyls (PCBs) in the diet. To evaluate this hypothesis, dioxins and related compounds were analysed in liver of female sea bass fed with a commercial or with a natural diet consisting of trash fish (bogue, Boops boops), and concentrations of vitellogenin (VTG) and 17β-estradiol (E2) were determined in plasma obtained previously in monthly samplings of these animals. As observed in other experiments, females fed with a commercial diet exhibited lower VTG and higher E2 plasma levels than females fed with the natural diet. In liver, sea bass fed with the commercial diet exhibited a profile clearly dominated by high-chlorinated dioxins while in fish fed with the natural diet this profile was dominated by low chlorinated furans. However, typical AhR ligands, such as 2,3,7,8-tetrachlorodibenzo- p-dioxin showed no differences between groups or, as is the case of planar PCBs, showed higher concentrations in the liver of fish fed with the natural diet. These results do not permit to explain the observed hormonal alterations by a possible antiestrogenic effect caused by dioxins and related compounds.

Effect of environmental contaminants on male reproduction

A substantial body of evidence has accumulated in recent years that consistently indicate various adverse effects of environmental contaminants on human health. Decreasing trend of male fertility in terms of sperm counts and sperm quality, along with other changes in male reproductive health, including congenital malformations and testicular cancer in humans, and similar problems in wildlife has been correlated to the exposure to environmental contaminants. Many environmental chemicals have been reported to cause these reproductive abnormalities by eliciting changes in endocrine control of reproduction; however the specific underlying mechanisms are poorly understood. The present review summarizes recent studies on environmental contaminants and associated possible mechanism leading to adverse effects on male reproduction. Numerous studies demonstrate the interaction of environmental toxicants with steroid receptors and thereby causing interference with developmental and functional aspects of testis, epididymis and accessory sex organs. Induction of reactive oxygen species (ROS) by environmental contaminants and associated oxidative stress also have role in defective sperm function and male infertility.