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Sexual isolation and cuticular hydrocarbon differences between Drosophila santomea and Drosophila yakuba.

Drosophila santomea and Drosophila yakuba are two sister species inhabiting Saõ Tomé island. Previous studies showed that both species display strong reproductive isolation, although they can produce a few viable hybrids. Our study tried to understand the mechanism of this ethological isolation between two allopatric strains. A strong sexual isolation was confirmed, with a marked asymmetry. Comparisons of latency times to either courtship or copulation suggest that males do not discriminate females, whereas D. yakuba females, but not D. santomea females, accept their homospecifics more quickly. Cuticular hydrocarbon compositions of both species and sexes were also established with gas chromatography (GC) and GC/mass spectrometry analysis. All have (Z)-7-tricosene as their major compound. There are several quantitative differences between species for few minor compounds. The largest difference concerns n-heneicosane, which is more abundant in D. santomea than in D. yakuba flies (up to seven times more between males). A similar quantitative difference was also found in a pair of sympatric strains. Furthermore, D. yakuba males artificially perfumed with n-heneicosane were discriminated negatively by D. yakuba females, suggesting a role for this compound in the sexual isolation between these two species.

Alkenes↗

Characterization of a female-produced courtship pheromone in the parasitoid Nasonia vitripennis.

Males of the parasitoid Nasonia vitripennis (Walker) (Hymenoptera: Pteromalidae) show a characteristic courtship behavior. We demonstrate that male arrestment and key behavioral elements of the courtship sequence are mediated by a female-derived contact sex pheromone. Males were arrested on paper disks treated with female extracts but not on those treated with male extracts. Male responsiveness was influenced by the surface to which female extracts were applied. Extracts applied to an extracted beetle elytron arrested males more strongly than those applied to filter paper of comparable size. However, more complex behavioral elements, such as head nodding and copulation attempts, were shown only when extracts were applied to extracted male cadavers, suggesting that tactile or visual cues synergize the male response. The chemicals involved are stable, of low volatility, and nonpolar. Dead females arrested males and elicited courtship behavior for at least 8 d. Males showed no sign of attraction to live females at a distance of 3 cm in an olfactometer. Fractionation of female extracts demonstrated that the activity was exclusively located in the nonpolar fraction. Analysis of the active fraction by gas chromatography-mass spectrometry revealed that cuticular hydrocarbons with chain lengths between 25 and 37 carbon units were present. Comparison of hydrocarbon profiles from males and females showed qualitative and quantitative differences. These results suggest that sex-specific cuticular hydrocarbons are the key signals mediating the male courtship behavior in N. vitripennis.

Animal Communication↗

The in vivo and in vitro effects of clomiphene citrate on ovulation, fertilization, and development of cultured mouse oocytes.

Clinical data indicate a high rate of discrepancy between ovulation and pregnancy rates and a high abortion rate in anovulatory women treated with clomiphene citrate. An animal model was used to examine the in vivo and in vitro effects of the drug on fertilization and preimplantation development. This work indicates that mice treated with clomiphene in the early follicular phase of pregnant mare's serum-induced cycles show a dose-dependent decrease in ovulated oocytes. No effect was seen on the further development of these ova. When the drug was administered in the late follicular phase, a dose-dependent decrease was seen in the rate of successful copulation, ovulation, and embryo development. Concomitant 17 beta-estradiol treatment reversed the copulatory, ovulatory, and developmental effects of clomiphene. A short in vitro treatment of oocytes with clomiphene prior to in vitro fertilization resulted in an increased proportion of degenerated and unfertilized ova. Moreover, this treatment caused a dose-dependent decrease in blastocyst formation.

Animals↗

Osteogenesis of the mandibular arch in Ts16 mice.

Males and females homozygous for the Robertsonian translocation specific for chromosomes 16 and 17 Rb(16.17)Bnr and male and females homozygous for the Robertsonian translocation for chromosomes 6 and 16 Rb(6.16)24Lub were bred to produce double heterozygotes [Rb(16.17)Bnr/Rb(6.16)24Lub]. Experimental data were based on 156 features: 70 euploid (control), 86 trisomic. Affected fetuses were identified by decreased size, shortened faces (flattened snouts), oedema, petechiae, open eyelids, and dysplastic ears. Identification of trisomics was substantiated by karyotyping the metaphasic spreads. Five gestational days were studied (14-18). Fetal age was assumed to be accurate as fertilization occurred within half an hour of copulation. Euploid specimens followed normal developmental paths of chondrogenesis, osteogenesis and related tissues. In trisomics, developmental faults increased unequally between gestational days 15 and 17: some tissues were mildly and others acutely affected. Among the trisomic disorders were diminished growth, lagging mitotic activity, and retarded and poorly ordered tissue development, especially of bone. All of these contributed to hypoplasia, hypocellularity, reduced vascular supply and enlarged intercellular spaces. Intensities of the mandibular abnormalities varied among litters and littermates. The severity of the developmental disorders of an individual Ts16 specimen differed among the tissue components studied. Of the trisomic mandibular tissues, bone was most frequently involved and Meckel's cartilage least.

Animals↗

The catecholamine-containing tubero-infundibular system and the control of luteinizing hormone release in the rabbit.

As determined by fluorescence histochemistry, the distribution of catecholamine-containing neurons in the hypothalamus of the female rabbit is similar to that seen in the rat. The fluorescence appearance of the hypothalamus was not appreciably different from normal at 0.25, 1, 4 or 24 h after mating, but in animals in which the synthesis of catecholamines was inhibited by the administration of the tyrosine hydroxylase inhibitor, H44/68, the number of fluorescent neurons seen in the nucleus periventricularis arcuatus following copulation was markedly reduced. However, the concentrations of luteinizing hormone (LH) in serum and in the pituitary glands of mated and unmated animals treated with H44/68 were not significantly different from those found in the corresponding control animals. This, together with the fact that ovulation followed mating in drug-treated rabbits suggests that a normal level of catecholamines in the tubero-infundibular system is not essential for the secretion of the amount of LH necessary for ovulation.

Animals↗

Heterosexual, autosexual and social behavior of adult male rhesus monkeys with medial preoptic-anterior hypothalamic lesions.

Bilateral radiofrequency lesions were made in the medial preoptic-anterior hypothalamic (MP-AH) area of 6 adult male rhesus monkeys; 5 sham-lesioned subjects served as controls. Behavioral analysis consisted of observations on copulatory behavior, yawning, masturbation and some aspects of social behavior. MP-AH lesions reduced or completely eliminated the display of manual contacts of the partner, mounts, intromissions and ejaculations without interfering with masturbation. Yawning, a sexually dimorphic behavior, was not affected either, Measure of several social behaviors indicated no evidence of social withdrawal or other aberrance of social interactions, which might have led to the decline in heterosexual behavior. The results with regard to copulatory behavior were consistent with the effects of MP-AH lesions in rats, cats and dogs. In rhesus monkeys it appears as though the MP-AH region is specifically involved in the mediation of heterosexual copulation and is not vital to the performance of other forms of male sexual activity such as masturbation. Also the MP-AH is not critical for the display of all sexually dimorphic behaviors. The types of behavioral change in MP-AH lesioned subjects differed to some extent from those following castration, indicating that the effects of the lesions cannot be explained as basically that of functional castration.

Animals↗

Dissociation of spontaneous and mating induced ovulation by frontal hypothalamic deafferentations in the rat.

Different types of anterior hypothalamic deafferentations have been used to investigate the nervous pathways involved in spontaneous and mating-induced ovulation in the rat. Knife cuts which circumscribed the suprachiasmatic nuclei on all but their ventral surface and either their rostral or caudal poles prevented spontaneous ovulation, but the rats were sexually receptive (copulation plugs and sperm in the smear), and mating induced ovulation. Similar types of cut extended dorsally so as to sever the continuity between the preoptic area and the mediobasal hypothalamus also prevented spontaneous ovulation, and although these rats were also receptive, mating did not induce ovulation. Two possible explanations are considered: either (i) that difference between the effects of the two types of cut is a direct function of the differing proportions of gonadotrophic hormone-containing axons severed, or (ii) cuts in the region of the suprachiasmatic nuclei specifically impair a mechanism for the maintenance of diurnal rhythms, to which the abnormality in gonadotrophin control is secondary.

Afferent Pathways↗

Sensory innervation of the external and internal genitalia of the female rat.

Using a whole-nerve recording method, the genitalia of the female rat were found to receive afferent innervation as follows. Pelvic nerve: vagina, cervix, and perineal skin; hypogastric nerve: cervix and proximal three fifths of the uterus; pudendal nerve: skin of perineum, inner thigh, and clitoral sheath. It is probable that the pudendal and pelvic nerves are activated during copulation, and that all 3 nerves are activated during parturition.

Action Potentials↗

Morphine and dynorphin(1-13) microinjected into the medial preoptic area and nucleus accumbens: effects on sexual behavior in male rats.

The effects on sexual behavior of opiate receptor stimulation within A10 and A14 terminal areas were examined in the following experiments. Morphine (0.01-6 nmol) and dynorphin(1-13) (0.01-3 pmol) were microinjected into the medial preoptic area (MPOA). Morphine (10-100 pmol) and dynorphin (10-100 fmol) injected into the MPOA reduced both the latency to ejaculate and the number of intromissions triggering ejaculation. Morphine (6 nmol) produced a failure to resume copulating following the second ejaculation. Morphine (1-10 nmol) injected into the nucleus accumbens (ACC) shortened the latency to the first intromission and lengthened the second postejaculatory interval. Naloxone (3 mg/kg i.p.) reversed the effects of morphine on intromission latency and attenuated the lowering of ejaculatory threshold.

Animals↗

Opioid modulation and sensitization of dopamine release elicited by sexually relevant stimuli: a high speed chronoamperometric study in freely behaving rats.

High speed chronoamperometry was used to measure oxidation and reduction currents associated with monoamine release in the caudate putamen and nucleus accumbens of freely moving rats exposed to bedding from cages that housed other male, ovariectomized female, or estradiol-progesterone-primed female rats. Estrus female, but not male or ovariectomized female bedding potently increased the electrochemical signal from electrodes implanted within the nucleus accumbens, and less effectively from more dorsal sites. Naloxone pretreatment attenuated the increase in the electrochemical signal. Repeated exposure to estrus female bedding led to an increased, or sensitized, response within the nucleus accumbens that was also sensitive to naloxone pretreatment. The ratios of the reduction to oxidation currents indicated that dopamine was the principal contributor to the increase in the electrochemical signal, suggesting that activation of the mesolimbic dopamine system accompanies exposure to sexually relevant stimuli. These results suggest that the facilitation of sexual behaviors by dopamine may be due, at least in part, to the processing of incentive motivational cues, and not necessarily to effects on copulation, itself. The results of the present study also suggest that opioid peptides contribute to the activation of mesolimbic dopamine by sexually relevant olfactory stimuli.

Animals↗

Sexual behavior increases c-fos expression in the forebrain of the male rat.

The ability of a wide variety of pharmacological and physiological stimuli to increase neuronal expression of Fos has led to the suggestion that it might serve as a marker of neuronal activation. Psychomotor stimulants increase the release of dopamine from the terminals of nigrostriatal and mesolimbic neurons and enhance Fos immunoreactivity in the striatum and nucleus accumbens (NAc). Because sexual behavior also increases dopamine release in these and other forebrain regions, the present study examined the effect of copulation on Fos immunoreactivity in the forebrain of intact, sexually active male rats. Sexual behavior produced a striking increase in Fos immunoreactivity in the medial preoptic area (MPOA), NAc, bed nucleus of the stria terminalis and piriform cortex. However, no increase in Fos immunoreactivity was observed in the striatum. These results are consistent with neurochemical, physiological, and behavioral data suggesting that the MPOA and NAc are important substrates of sexual behavior.

Animals↗

Mating activates androgen receptor-containing neurons in chemosensory pathways of the male Syrian hamster brain.

Fos-immunoreactivity is induced during mating in the male Syrian hamster in limbic areas that relay chemosensory information and contain receptors for gonadal steroid hormones. The induction of Fos is an index of neuronal activation. After mating, c-fos expression is greatest in subnuclei of the medial amygdaloid nucleus (Me), bed nucleus of the stria terminalis (BNST), and medial preoptic area (MPOA). The present study determined if individual neurons in these activated subnuclei contain androgen receptors. We aim to understand how essential chemosensory and hormonal signals are integrated to control copulation. Adult male hamsters (n = 6) were allowed to mate with a sexually receptive female for 30 min. They were perfused 1 h later with 4% paraformaldehyde and 40 microns frozen sections were processed for immunocytochemistry using antisera against Fos (Cambridge Research Biochemicals) and the androgen receptor (G.S. Prins). The brains of three non-mated males were also processed for Fos immunocytochemistry. Mating significantly increased the number of Fos-immunoreactive neurons within subnuclei of Me, BNST, and MPOA relative to non-mated males (P < 0.05). These nuclei contained abundant androgen receptors. In the corticomedial amygdala, 20-40% of Fos-immunoreactive neurons in mated hamsters expressed androgen receptors. Although few androgen receptors are found in the anteromedial and postero-intermediate subdivisions of the BNST, these areas exhibited 26% and 47% co-localization, respectively. In posteromedial BNST, which contains large numbers of steroid receptor-containing neurons, androgen receptors were identified in 48% of Fos-immunoreactive neurons. In the MPOA, 54% of Fos-immunoreactive neurons expressed the androgen receptor throughout the rostrocaudal extent of the medial preoptic nucleus (MPN).(ABSTRACT TRUNCATED AT 250 WORDS)

Amygdala↗

Pre- and post-translational regulation of aromatase by steroidal and non-steroidal aromatase inhibitors.

Treatment of castrated quail with testosterone (T) reliably activates male copulatory behavior and, at the same time, increases the aromatase activity (AA), the number of aromatase-immunoreactive (ARO-ir) cells and the concentration of aromatase mRNA as measured by RT-PCR in the brain. All these effects can be mimicked by estrogens. The behavioral effects of T can be blocked by a variety of aromatase inhibitors and, in parallel, the AA is strongly inhibited in the preoptic area (POA). We showed recently that the steroidal inhibitor, 4-OH-androstenedione (OHA) markedly decreases the immunostaining density of brain ARO-ir cells while the non-steroidal inhibitor, R76713 (racemic Vorozole; VOR) unexpectedly increased the density of this staining, despite the fact that the enzyme activity was completely inhibited. To generalize these findings and try to identify the underlying mechanism, we compared here the effects of two steroidal (OHA and androstatrienedione [ATD]) and two non-steroidal (VOR and Fadrozole [FAD]) aromatase inhibitors on the aromatase immunostaining and aromatase mRNA concentration in the brain of castrated quail concurrently treated with T. The 4 inhibitors significantly blocked the activation by T of male copulation. The two steroidal inhibitors decreased the immunostaining of brain ARO-ir cells but both VOR and FAD markedly enhanced the density of this staining. In parallel, OHA and ATD completely blocked the T-induced increase in aromatase mRNA concentration, while VOR and FAD had no effect on these RNA concentrations in the POA-anterior hypothalamus and they decreased them only slightly in the posterior hypothalamus. Taken together these results suggest that the inhibition of AA by ATD or OHA and the subsequent removal of locally produced estrogens blocks the synthesis of aromatase presumably at the transcriptional level. By contrast, the two non-steroidal inhibitors tested here block AA but in parallel increase the aromatase immunostaining. This effect does not result from an enhanced transcription and it is therefore speculated that these compounds increase either the translation of the aromatase mRNA or the half-life of the protein itself.

Animals↗

Biological, microscopic and scanning electron microscopic investigations of the effects of postinor /d-norgestrel/ in rabbits.

The contraceptive effect of d-norgestrel given immediately after copulation in various quantities was investigated in rabbits. It was established that the effect is dose-dependent. A correlation was found between the amount of dose administered and the changes taking place on the surface of endometrium. It can be suggested that d-norgestrel alters the surface of the endometrium to such an extent that nidation is unable to take place; therefore, it can be used for postcoital contraception at any time.

Animals↗

Levonorgestrel as a postcoital contraceptive.

Time-release pellets of levonorgestrel (LNG), the progestogenic hormone contained in the contraceptive system Norplant, were implanted subdermally in mice, after the animals had mated and ovulated but before uterine implantation of embryos would have occurred, to examine whether the hormone could reduce the number of embryos that subsequently implanted and, if so, when it had to be administered in the postcoital period to achieve that effect. Hybrid female mice (C57BL x CBA) were paired with breeder males (CD-1) and LNG pellets were implanted on day 0, the day on which copulation plugs were found, or on day 2 or day 3 in the postcoital period. Mice in some groups were sacrificed on day 14 of the gestation period, and numbers of fetuses and/or resorption sites were counted, while mice in other groups were allowed to go to term. When LNG pellets were implanted subdermally on day 0 of the postcoital period, pellets designed to release 1.5 mg of hormone in 21 days failed to exhibit a contraceptive effect, but pellets designed to release 5 mg of hormone in 90 days were totally effective in preventing uterine implantation of embryos. Although the 5 mg pellets did not prevent embryos from implanting in all cases when administered on day 2, they prevented pregnancies from going to term by causing resorption of those embryos that did implant. When the pellets were implanted as late as day 3 in the postcoital period, uterine implantation of embryos occurred and fetuses were carried to term. Results of the study indicate that subdermal implants of LNG inserted postcoitally can prevent uterine implantation of embryos in mice, and thereby prevent pregnancy, despite fertilization of oocytes having occurred, if the hormone implants are inserted before day 3 of the postcoital period.

Animals↗

Reproductive function in rats after mifepristone-induced termination of pregnancy.

The effect of mifepristone, a potent progesterone receptor antagonist, on the reproductive function during early pregnancy in rats was examined. A single dose of this drug (10 mg/kg) was injected s.c. at 1200 h on day 4 (Group 1), day 7 (Group II) or day 10 (Group III) of pregnancy. Gestation was interrupted and the vaginal cytology showed a typical proestrus condition two days after mifepristone treatment in all groups. When compared with cycling proestrus rats, serum LH concentrations at 1800 h in the mifepristone-induced proestrus were lower in Group I, similar in Group II and higher in Group III, and serum prolactin (PRL) values were lower in Group I, but not different in Groups II and III. Serum progesterone levels were higher in the three experimental groups when compared with cycling proestrus rats, and similar to that of pregnant rats. Rats in Group I showed a significantly lower sexual receptivity and ovulation rate when compared with Groups II and III or cycling proestrus rats. Most of the mifepristone-treated rats that copulated during the night of the induced proestrus did not become pregnant and showed a delayed pseudopregnancy-like condition. These results indicate that mifepristone administered in a single dose to early pregnant rats terminates pregnancy and induces a proestrus condition two days after treatment followed by successful postovulatory contraception. The mifepristone-induced proestrus is characterized by a differential pattern of serum LH, PRL and progesterone concentrations, mating behavior and ovulation rates, depending on the day of pregnancy when mifepristone is administered.

Abortifacient Agents, Steroidal↗

Structure, cell-specific expression, and mating-induced regulation of a Drosophila melanogaster male accessory gland gene.

The accessory gland of male insects is a secretory tissue of the genital tract made up of several distinct cell types. It secretes components of the ejaculatory fluid which have an important effect on the postmating behavior of the female. We have examined the sequence, structure, and expression of a gene, mst 316, expressed exclusively in the accessory glands of male Drosophila melanogaster. The mst 316 RNA encodes a small, basic protein of 52 amino acids that exhibits features common to precursors of secreted peptides, including a hydrophobic N-terminus. The tissue-specific expression of the mst 316 gene was studied using an mst 316--lacZ hybrid gene inserted into Drosophila by germ line transformation. The mst 316-lacZ fusion protein is expressed exclusively in the "main" cells of the accessory gland. It is first detected upon eclosion and exhibits a burst of synthesis in the first 3 days of adult life. The synthesis of the fusion protein is stimulated by mating, so that beta-galactosidase activity levels are two- to sixfold higher in males allowed to copulate with females compared to virgin male controls of the same age. The mating-stimulated synthesis of the mst 316-lacZ fusion protein, and by inference of the native gene product, appears to be due at least in part to increased transcript levels.

Amino Acid Sequence↗

Hippocampal EEG and behaviour in dog. III. Hippocampal EEG correlates of stimulus-response tasks and of sexual behaviour.

A dog was trained to perform a spatial sound discrimination. The hippocampal EEG correlates and the movement correlates of correct trials were compared with those of incorrect trials and of 'pressings in between'. Correct and wrong responses on a place learning task were compared both with respect to the hippocampal EEG correlates and to the motor behaviour. Where significant differences in EEG correlates were found, the motor behaviour differed significantly as well. The same was found for 2 behavioral states with different motor behavioural qualities which occurred during copulation. The data support the hypothesis that the spectral properties of the hippocampal EEG reflect in a predictable but non-specific way the intensity of motor behaviour. They question the postulated correlations between hippocampal EEG and 'internal processes', on the ground that unambiguous experimental verification of such hypothesis appears to be very difficult.

Animals↗