Hair from sexually active bucks strongly activates olfactory sensory inputs but fails to trigger early first ovulation in prepubescent does.

Early exposure of does to sexually active bucks triggers early puberty onset correlating with neuroendocrine changes. However, the sensory pathways that are stimulated by the male are still unknown. Here, we assessed whether responses to olfactory stimuli are modulated by social experience (exposure to males or not) and/or endocrine status (prepubescent or pubescent). We used a calcium imaging approach on goat sensory cells from the main olfactory epithelium (MOE) and the vomeronasal organ (VNO). For both cell types, we observed robust responses to active male hair in females under three physiological conditions: prepubescent females isolated from males (ISOL PrePub), pubescent females exposed to males (INT Pub) and isolated females (ISOL Pub). Response analysis showed overall greater proportion of responses to buck hair in ISOL PrePub. We hypothesized that females would be more responsive to active buck hair during the prepubertal period, with numerous responses perhaps originating from immature neurons. We also observed a greater proportion of mature olfactory neurons in the MOE and VNO of INT Pub females suggesting that male exposure can induce plastic changes on olfactory cell function and organization. To determine whether stimulation by male odor can advance puberty, we exposed prepubescent does to active buck hair (ODOR). In both ODOR and females isolated from males (ISOL) groups, puberty was reached one month after females exposed to intact bucks (INT), suggesting that olfactory stimulation is not sufficient to trigger puberty.

Male-induced early puberty correlates with the maturation of arcuate nucleus kisspeptin neurons in does.

In goats, early exposure of spring-born females to sexually active bucks induces an early puberty onset assessed by the first ovulation. This effect is found when females are continuously exposed well before the male breeding season starting in September. The first aim of this study was to evaluate whether a shortened exposure of females to males could also lead to early puberty. We assessed the onset of puberty in Alpine does isolated from bucks (ISOL), exposed to wethers (CAS), exposed to intact bucks from the end of June (INT1), or mid-August (INT2). Intact bucks became sexually active in mid-September. At the beginning of October, 100% of INT1 and 90% of INT2 exposed does ovulated, in contrast to the ISOL (0%) and CAS (20%) groups. This demonstrated that contact with males that become sexually active is the main factor prompting precocious puberty in females. Furthermore, a reduced male exposure during a short window before the breeding season is sufficient to induce this phenomenon. The second aim was to investigate the neuroendocrine changes induced by male exposure. We found a significant increase in kisspeptin immunoreactivity (fiber density and number of cell bodies) in the caudal part of the arcuate nucleus of INT1 and INT2 exposed females. Thus, our results suggest that sensory stimuli from sexually active bucks (e.g., chemosignals) may trigger an early maturation of the ARC kisspeptin neuronal network leading to gonadotropin-releasing hormone secretion and first ovulation.

Population density does not affect seasonal regulation of reproductive physiology in male water voles.

Most small rodent species display cyclic fluctuations in their population density. The mechanisms behind these cyclical variations are not yet clearly understood. Density-dependent effects on reproductive function could affect these population variations. The fossorial water vole ecotype, Arvicola terrestris, exhibits multi-year cyclical dynamics with outbreak peaks. Here, we monitored different water vole populations over 3 years, in spring and autumn, to evaluate whether population density is related to male reproductive physiology. Our results show an effect of season and inter-annual factors on testis mass, plasmatic testosterone level, and androgen-dependent seminal vesicle mass. By contrast, population density does not affect any of these parameters, suggesting a lack of modulation of population dynamics by population density.

Exposure of rams in sexual rest to sexually activated males in spring increases plasma LH and testosterone concentrations.

Eight stimulating rams, and twelve stimulated rams, were used to determine whether a similar endocrine response to the introduction of sexually active males in spring in a flock of ewes is observed in a flock of rams. The stimulating rams (n = 4) were induced into a sexually active state by exposure to 2 months of long days (16 h light/d) (15 December-15 February). At the end of the long-day period, rams were returned to the natural photoperiod. Control-stimulating rams (n = 4) were kept under the natural photoperiod. On April 20, stimulated rams were divided into 2 groups, and joined with activated (ACT; n = 6) or control stimulating rams (C; n = 6). On the day of ram introduction, stimulated rams were blood sampled for 8 h at 20-min intervals, from 4 h before to 4 h after ram introduction, and next day from 24 to 28 h after ram introduction, and analyzed for plasma LH concentrations, and 10, 20 and 30 days after ram introduction to measure plasma testosterone levels. Mean (±SEM) plasma LH concentrations (ng/ml) of stimulated rams were similar during the 4 h before stimulating-ram introduction (ACT: 0.59 ± 0.03; C: 0.53 ± 0.04; P > 0.05). The introduction of the photoperiod-treated stimulating rams increased LH concentrations of stimulated rams during the 4 h after their introduction (1.14 ± 0.37) compared with the C group (0.51 ± 0.03; P < 0.05), especially during the first hour (ACT: 0.93 ± 0.16; C: 0.49 ± 0.03; P < 0.05), and during the blood sampling period 24-28 h after ram introduction (0.75 ± 0.07 vs. 0.58 ± 0.04; P < 0.05). Before the introduction of stimulating rams, the LH pulse frequencies and amplitudes did not differ between groups; however, LH pulsatility was higher at 4 h (0.58 ± 0.11 pulses/h; P < 0.05), and had trend to be higher 24 h (0.50 ± 0.06) (P = 0.10) after the introduction of the photoperiod-treated stimulating rams compared with the control-stimulating rams (0.29 ± 0.08 and 0.29 ± 0.10, respectively). As for LH pulses, there was an effect of group (P < 0.05) on LH amplitude, which presented a trend to be higher in ACT rams 4 h after ram introduction (1.68 ± 0.30; P < 0.10) and higher 24 h (1.07 ± 0.08; P < 0.05) after ram introduction, compared with LH amplitudes of C rams (0.71 ± 0.06 and 0.82 ± 0.07, respectively). Plasma testosterone concentrations of rams exposed to photoperiod-treated activated rams were higher than those of rams exposed to control-stimulating rams, at 4 h, 20 and 30 days after ram introduction (P < 0.05). In conclusion, sexually active rams in spring are able to stimulate LH and testosterone secretion of other rams in sexual rest, a phenomenon we called "ram-to-ram effect".

Photoperiod is involved in the regulation of seasonal breeding in male water voles (Arvicola terrestris).

Mammals living at temperate latitudes typically display annual cyclicity in their reproductive activity: births are synchronized when environmental conditions are most favorable. In a majority of these species, day length is the main proximate factor used to anticipate seasonal changes and to adapt physiology. The brain integrates this photoperiodic signal through key hypothalamic structures, which regulate the reproductive axis. In this context, our study aimed to characterize regulations that occur along the hypothalamo-pituitary-gonadal (HPG) axis in male fossorial water voles (Arvicola terrestris, also known as Arvicola amphibius) throughout the year and to further probe the implication of photoperiod in these seasonal regulations. Our monthly field monitoring showed dramatic seasonal changes in the morphology and activity of reproductive organs, as well as in the androgen-dependent lateral scent glands. Moreover, our data uncovered seasonal variations at the hypothalamic level. During the breeding season, kisspeptin expression in the arcuate nucleus (ARC) decreases, while RFRP3 expression in the dorsomedial hypothalamic nucleus (DMH) increases. Our follow-up laboratory study revealed activation of the reproductive axis and confirmed a decrease in kisspeptin expression in males exposed to a long photoperiod (summer condition) compared with those maintained under a short photoperiod (winter condition) that retain all features reminiscent of sexual inhibition. Altogether, our study characterizes neuroendocrine and anatomical markers of seasonal reproductive rhythmicity in male water voles and further suggests that these seasonal changes are strongly impacted by photoperiod.

Field study reveals morphological and neuroendocrine correlates of seasonal breeding in female water voles, Arvicola terrestris.

Seasonally breeding mammals display timely physiological switches between reproductive activity and sexual rest, which ensure synchronisation of births at the most favourable time of the year. These switches correlate with seasonal changes along the hypothalamo-pituitary-gonadal axis, but they are primarily orchestrated at the hypothalamic level through environmental control of KISS1-dependent GnRH release. Our field study shows that births of fossorial water voles, Arvicola terrestris, are concentrated between March and October, which indicates the existence of an annual reproductive cycle in this species. Monthly field monitoring for over a year further reveals dramatic seasonal changes in the morphology of the ovary, uterus and lateral scent glands, which correlate with the reproductive status. Finally, we demonstrate seasonal variation in kisspeptin expression within the hypothalamic arcuate nucleus. Altogether, this study demonstrates a marked rhythm of seasonal breeding in the water vole and we speculate that this is governed by seasonal changes in photoperiod.

Light-induced sexually active rams provoke LH preovulatory surges and enhances LH concentrations in ewes after progestagen treatment.

The effect of the introduction of sexually active rams in spring on LH secretion in ewes after progestagen treatment has been studied. Two rams were induced into a sexually active state by exposure to 2 months of long days (16 h of light/d) from 1 January (SAR), and another 2 rams were exposed to the natural photoperiod, so that they were not sexually activated in spring (control; C). At the end of the long-day period, rams were returned to natural photoperiod conditions. Fifteen ewes synchronized in estrus by intravaginal sponges were assigned to three groups at sponge withdrawal (hour 0): SAR (n = 5), exposed to SAR rams; C (n = 5), exposed to C rams, and ISO (n = 5), kept isolated from rams. Twenty-four hours after pessary removal (hour 0), rams were introduced into the SAR and C groups. Three SAR ewes presented preovulatory LH surges; the proportion was significantly (P < 0.05) higher in this group (3/5) than in the others (C: 0/5, ISO: 0/5). SAR introduction induced a more marked (P < 0.05) increase in mean LH plasma concentrations (before rams: 1.34 ± 0.19; after rams: 6.94 ± 2.66 ng/ml) than C (before: 0.96 ± 0.29; after: 3.60 ± 1.44) or ISO (before: 1.26 ± 0.42; after: 2.14 ± 1.36) groups, and significantly (P < 0.05) higher plasma LH levels after ram introduction. In conclusion, only light-treated sexually activated rams induced LH preovulatory surges in ewes in the seasonal anestrus, when ewes are synchronized with progestagen treatment in the absence of eCG.

In vivo magnetic resonance imaging reveals the effect of gonadal hormones on morphological and functional brain sexual dimorphisms in adult sheep.

Sex differences in the brain and behavior are produced by the perinatal action of testosterone, which is converted into estradiol by the enzyme aromatase in the brain. Although magnetic resonance imaging (MRI) has been widely used in humans to study these differences, the use of animal models, where hormonal status can be properly manipulated, is necessary to explore the mechanisms involved. We used sheep, a recognized model in the field of neuroendocrinology, to assess brain morphological and functional sex differences and their regulation by adult gonadal hormones. To this end, we performed voxel-based morphometry and a resting-state functional MRI approach to assess sex differences in gonadally intact animals. We demonstrated significant sex differences in gray matter concentration (GMC) at the level of the gonadotropic axis, i.e., not only within the hypothalamus and pituitary but also within the hippocampus and the amygdala of intact animals. We then performed the same analysis one month after gonadectomy and found that some of these differences were reduced, especially in the hypothalamus and amygdala. By contrast, we found few differences in the organization of the functional connectome between males and females either before or after gonadectomy. As a whole, our study identifies brain regions that are sexually dimorphic in the sheep brain at the resolution of the MRI and highlights the role of gonadal hormones in the maintenance of these differences.

Sexually active bucks are a critical social cue that activates the gonadotrope axis and early puberty onset in does.

In rodents, early exposure to adult male is well known to induce an early puberty in females (Vandenbergh effect). This phenomenon has been less studied in other mammals. In goats, despite our extensive knowledge about the "male-effect" phenomenon in adults (i.e. ovulation induced by the introduction of the male during the anestrous), there are few data on the consequences of an early exposure of females to males. Here, we evaluated the puberty onset of young alpine goats when raised since weaning with intact bucks (INT), with castrated bucks (CAS) or isolated from bucks (ISOL). The INT group had the first ovulation 1.5 month before the two other groups. Despite the earlier puberty the INT group of females had normal and regular ovarian cycles. Morphological study of the genital tract showed that at 6 months, uterus of INT goats was 40% heavier than CAS and ISOL goats. Moreover, INT females had a myometrium significantly thicker and INT was the only group having corpora lutea. In our study, INT females were pubescent in the month following the entry of bucks into the breeding season, suggesting that only sexually active bucks provide the signal responsible for puberty acceleration. By removing direct contact with the bucks, we showed that somatosensory interactions were dispensable for an early puberty induction. Finally, no difference in the GnRH network (fiber density and number of synaptic appositions) can be detected between pubescent and non-pubescent females, suggesting that the male stimulations triggering puberty onset act probably on upstream neuronal networks, potentially on kisspeptin neurons.

Exposure to Photoperiod-Melatonin-Induced, Sexually-Activated Rams after Weaning Advances the Resumption of Sexual Activity in Post-Partum Mediterranean Ewes Lambing in January.

This study was aimed to determine whether the presence of sexually stimulated rams by photoperiodic and melatonin treatments can advance the resumption of post-partum sexual activity in Mediterranean ewes lambing in January and weaned at the end of the breeding season at 41°N, in March. Rams were exposed to two months of long days (16 h light/day) and given three melatonin implants at the end of the long days (sexually-activated rams; SAR). Control rams (CR) were exposed to the natural photoperiod. Thirty-six ewes weaned on 25 February were assigned to one of two groups. From 1 March to 30 June, one group was housed with four SAR males (SAR-treated; n = 18), and the other group (CR-treated; n = 18) was housed with four unstimulated rams. Ovulation was assessed once per week based on plasma progesterone concentrations. Estrus was monitored daily by marks left on ewes by rams' harnesses. SAR-treated ewes had a shorter (p < 0.01) weaning-first estrus interval than CR-treated ewes (61 ± 17 days vs. 102 ± 47 days; mean date of first estrus after weaning on April 26 and June 6, respectively). The proportion of the ewes ovulating in April or May was higher (p < 0.05) in the SAR-treated group than in the CR-treated group. SAR-treated ewes resumed estrous activity sooner than CR-treated ewes such that, in April, May, and June, the proportion of females that exhibited estrus was higher (p < 0.01) in the SAR-treated group (72%, 89%, and 100%, respectively) than in the CR-treated group (17%, 44%, and 61%, respectively). In conclusion, the introduction at weaning of sexually activated rams advanced the resumption of estrous activity in ewes in spring. The practical implications of this work could be important in ewes adapted for intensive production and accelerated lambing systems.

Computation of a high-resolution MRI 3D stereotaxic atlas of the sheep brain.

The sheep model was first used in the fields of animal reproduction and veterinary sciences and then was utilized in fundamental and preclinical studies. For more than a decade, magnetic resonance (MR) studies performed on this model have been increasingly reported, especially in the field of neuroscience. To contribute to MR translational neuroscience research, a brain template and an atlas are necessary. We have recently generated the first complete T1-weighted (T1W) and T2W MR population average images (or templates) of in vivo sheep brains. In this study, we 1) defined a 3D stereotaxic coordinate system for previously established in vivo population average templates; 2) used deformation fields obtained during optimized nonlinear registrations to compute nonlinear tissues or prior probability maps (nlTPMs) of cerebrospinal fluid (CSF), gray matter (GM), and white matter (WM) tissues; 3) delineated 25 external and 28 internal sheep brain structures by segmenting both templates and nlTPMs; and 4) annotated and labeled these structures using an existing histological atlas. We built a quality high-resolution 3D atlas of average in vivo sheep brains linked to a reference stereotaxic space. The atlas and nlTPMs, associated with previously computed T1W and T2W in vivo sheep brain templates and nlTPMs, provide a complete set of imaging space that are able to be imported into other imaging software programs and could be used as standardized tools for neuroimaging studies or other neuroscience methods, such as image registration, image segmentation, identification of brain structures, implementation of recording devices, or neuronavigation. J. Comp. Neurol. 525:676-692, 2017. © 2016 Wiley Periodicals, Inc.

A high level of male sexual activity is necessary for the activation of the medial preoptic area and the arcuate nucleus during the 'male effect' in anestrous goats.

In small ungulates such as sheep or goats, the introduction of a male among a group of anovulatory females during the anestrus season leads to the reactivation of the gonadotrope axis and ovulation, a phenomenon known as the 'male effect'. In goats, our previous studies have demonstrated the importance of male sexual activity for an efficient reactivation of the gonadotrope axis assessed through ovulation and blood LH pulsatility. In the present experiment, we assessed whether the level of male sexual activity would also induce differential activation of two brain regions of key importance for the reactivation of GnRH activity, namely the medial preoptic area and the hypothalamic arcuate nucleus. In both structures, we observed a differential activation of Fos in females, depending on the level of buck sexual activity. Indeed, goats unexposed to males showed low levels of expression of Fos while those exposed to sexually inactive bucks showed an intermediate level of Fos expression. Finally, the highest level of Fos expression was found in females exposed to sexually active males. However, and contrary to our initial hypothesis, we were not able to find any specific activation of kisspeptin cells in the arcuate nucleus following the introduction of highly sexually active males. As a whole, these results demonstrate that the level of male sexual activity is a key factor to stimulate brain regions involved in the control of the gonadotrope axis in the context of the male effect in goats.

Sexually active bucks are efficient to stimulate female ovulatory activity during the anestrous season also under temperate latitudes.

Goats are seasonal breeders and photoperiod is the main cue controlling the onset and offset of the breeding season. Nevertheless introducing a sexually active buck in a group of females during anestrous can stimulate their reproductive function and induce ovulation. This "male-effect" is very efficient under subtropical latitudes, when using sexually active males previously stimulated by a photoperiodic treatment. However, there is less evidence of its feasibility under temperate latitudes where the more important variation in day length could be responsible for a stronger inhibition of female sexual activity. The aim of this study was therefore to determine whether intense sexual activity can be induced in alpine bucks during the non-breeding season by a long-day treatment under temperate latitude and if these males could be used to produce an efficient male-effect. Bucks (n=21) were divided in two groups, one submitted to a photoperiodic treatment from November 1st to January 15th and then switched to natural photoperiod, while the other group remained entirely under the natural photoperiod. The ones submitted to this light treatment exhibit higher testicular volume and testosterone level 6 weeks after the end of the treatment. At the end of March, bucks were used to stimulate anestrous does (n=41) continuously for 15 days. We showed that (a) light treatment was efficient to induce an increase of sexual activity in bucks and (b) that the introduction of stimulated bucks among females induced a significantly higher proportion of ovulation in anestrous does than control bucks (86% vs 5%). Our results indicate that under temperate latitudes induction of ovulation in females during the anestrous season is feasible using bucks treated with long-days during winter.

One day of contact with photostimulated bucks is sufficient to induce ovulation in seasonally anestrous goats.

In seasonally anovulatory goats, daily contact with photostimulated bucks for 15 days stimulates ovulations in most females. In this study, we determined whether ovulation could be induced in goats exposed to photostimulated bucks for less than 15 days. Bucks were rendered sexually active during the nonbreeding season by exposure to 2.5 months of long days from November 1. The control group of females was exposed to one photostimulated buck for 15 days (n = 12). Other three experimental groups were exposed to males (n = 1 per group) for 1, 5, or 10 days (n = 14 or 15 females per group). Ovulations were determined by measurement of daily plasma progesterone concentrations during 17 days. All females from the control and experimental groups ovulated at least once during the experiment (P > 0.05). Furthermore, the proportions of goats that displayed a short luteal phase followed by a new ovulation, or a normal luteal phase after being in contact with males, did not differ between groups depending on the duration of time of contact with the photostimulated males (P > 0.05). In contrast, the proportions of females that displayed a short luteal phase followed by anovulation were greater in goats in contact with males for 1 day than in those in contact with males for 10 and 15 days (P < 0.05), whereas they did not differ from females exposed to males for 5 days (P > 0.05). We conclude that 1 day of contact with sexually active males is long enough to stimulate the ovulatory activity in seasonally anovulatory goats. However, a significantly higher proportion of females exposed to males for 1 day did not ovulate again after showing a short luteal phase.

Enhancing bull sexual behavior using estrus-specific molecules identified in cow urine.

Deficiencies in bull mating behavior have implications for bovine artificial insemination activities. The aim of this study was to identify the compounds present in fluids emitted by cows during estrus, which could enhance bull libido. Chemical analysis of urine samples from cows led to the characterization of molecules varying specifically at the preestrous and estrous stages. The synthetic counterpart molecules (1,2-dichloroethylene, squalene, coumarin, 2-butanone, oleic acid) were used to investigate the biological effects on male sexual behavior and sperm production. When presented to males, 2-butanone and oleic acid synthetic molecules significantly lowered mounting reaction time and ejaculation time (-33% and 21% after 2-butanone inhalation, respectively, P < 0.05). The "squalene +1,2-dichloroethylene" combination induced a 9% increase of sperm quantity (P < 0.05). This study suggests that the identified estrous-specific molecules could be part of the chemical signals involved in male and female mating behavior and may be used for a wide range of applications. The identification of these molecules may have implications for the cattle breeding industry.

Looking for the bird Kiss: evolutionary scenario in sauropsids.

BACKGROUND: The neuropeptide Kiss and its receptor KissR are key-actors in the brain control of reproduction in mammals, where they are responsible for the stimulation of the activity of GnRH neurones. Investigation in other vertebrates revealed up to 3 Kiss and 4 KissR paralogs, originating from the two rounds of whole genome duplication in early vertebrates. In contrast, the absence of Kiss and KissR has been suggested in birds, as no homologs of these genes could be found in current genomic databases. This study aims at addressing the question of the existence, from an evolutionary perspective, of the Kisspeptin system in birds. It provides the first large-scale investigation of the Kisspeptin system in the sauropsid lineage, including ophidian, chelonian, crocodilian, and avian lineages. RESULTS: Sauropsid Kiss and KissR genes were predicted from multiple genome and transcriptome databases by TBLASTN. Phylogenetic and syntenic analyses were performed to classify predicted sauropsid Kiss and KissR genes and to re-construct the evolutionary scenarios of both gene families across the sauropsid radiation.Genome search, phylogenetic and synteny analyses, demonstrated the presence of two Kiss genes (Kiss1 and Kiss2 types) and of two KissR genes (KissR1 and KissR4 types) in the sauropsid lineage. These four genes, also present in the mammalian lineage, would have been inherited from their common amniote ancestor. In contrast, synteny analyses supported that the other Kiss and KissR paralogs are missing in sauropsids as in mammals, indicating their absence in the amniote lineage. Among sauropsids, in the avian lineage, we demonstrated the existence of a Kiss2-like gene in three bird genomes. The divergence of these avian Kiss2-like sequences from those of other vertebrates, as well as their absence in the genomes of some other birds, revealed the processes of Kiss2 gene degeneration and loss in the avian lineage. CONCLUSION: These findings contribute to trace back the evolutionary history of the Kisspeptin system in amniotes and sauropsids, and provide the first molecular evidence of the existence and fate of a Kiss gene in birds.

Male sexual behavior contributes to the maintenance of high LH pulsatility in anestrous female goats.

The objective of this study was to determine the importance of male sexual behavior in stimulating LH secretion in anovulatory female goats. Two groups of females (n=10 per group) were each exposed to a buck in sexual rest and submitted to natural daylength. In one group, the buck was awake, whereas in the other group, it was sedated to prevent its sexual behavior. Two other groups of goats (n=10 per group) were exposed to sexually active bucks that had been exposed to 2.5 months of long days. In one group, the buck was awake, and in the other group, it was sedated. LH secretion was determined every 15 min from 4 h before introducing the bucks to 8 h after, then every 15 min again from 20 to 24 h after introducing the bucks. The bucks submitted to natural daylength did not stimulate LH secretion (P>0.05), whether they were sedated or not. In contrast, both the awake and the sedated light-treated bucks induced an increase (P<0.05) of LH pulsatility in the first 4 h following their introduction. However, pulsatility remained elevated until 24 h in the females exposed to the light-treated awake buck, whereas in the group with the light-treated sedated buck, pulsatility diminished (P<0.05) after the first 4 h of stimulation by the buck. In conclusion, the sexual behavior of males contributes to the maintenance of a high LH pulsatility up to 24 h after introduction into a group of anovulatory goats.

Phosphorylation of native porcine olfactory binding proteins.

The identification of various isoforms of olfactory binding proteins is of major importance to elucidate their involvement in detection of pheromones and other odors. Here, we report the characterization of the phosphorylation of OBP (odorant binding protein) and Von Ebner's gland protein (VEG) from the pig, Sus scrofa. After labeling with specific antibodies raised against the three types of phosphorylation (Ser, Thr, Tyr), the phosphate-modified residues were mapped by using the beta-elimination followed by Michael addition of dithiothreitol (BEMAD) method. Eleven phosphorylation sites were localized in the pOBP sequence and nine sites in the VEG sequence. OBPs are secreted by Bowman's gland cells in the extracellular mucus lining the nasal cavity. After tracking the secretion pathway in the rough endoplasmic reticulum of these cells, we hypothesize that these proteins may be phosphorylated by ectokinases that remain to be characterized. The existence of such a regulatory mechanism theoretically increases the number of OBP variants, and it suggests a more specific role for OBPs in odorant coding than the one of odorant solubilizer and transporter.

Modulation of estrogen receptors during development inhibits neurogenesis of precursors to GnRH-1 neurones: in vitro studies with explants of ovine olfactory placode.

The aim of the present study was to explore the putative effects of agonists and antagonists of the estradiol receptor on the early phase of GnRH-1 neuron development. To address this question we used an in vitro model of GnRH-1 neurons using cultured olfactory placode from sheep embryos on day 26 of gestation. Previous studies on this model have shown that in vitro the development of GnRH-1 neurons mimics in vivo development up to the start of pulsatile GnRH-1 secretion, To address the effects of modulating the estrogen receptor, cultures were treated with the endogenous and synthetic ligands of estradiol receptors: 17beta-estradiol, 17alpha-estradiol and tamoxifen. Neurogenesis was measured by incorporation of [(3)H]-thymidine. Morphometric parameters were evaluated by image analysis. The main results are that antagonism of estradiol receptors induced an important decrease in neurogenesis but had little effect on morphometric parameters, suggesting that during this early phase of development, maternal estrogens are important to achieve correct development of the GnRH-1 neuronal network.

High fertility using artificial insemination during deep anoestrus after induction and synchronisation of ovulatory activity by the "male effect" in lactating goats subjected to treatment with artificial long days and progestagens.

The response to the male effect was studied in two Saânen and two Alpine flocks over 5 consecutive years. Adult male and female goats were exposed to artificial long days (16h light and 8h darkness, 16L:8D) in open barns for approximately 3 months (between December 1 and April 15) followed by a natural photoperiod. Goats were treated for 11 days with fluorogestone acetate (FGA) or progesterone (CIDR) immediately before joining. Bucks carrying marking harnesses with adapted aprons joined females 49-63 days after the end of the long-day treatment (between April 30 and June 5) and were left with them for 5 days. In experiment 1 (n=142), FGA- and CIDR-treated goats were inseminated at a time based on the detection of oestrus. Two insemination groups were distinguished by the occurrence of marking over a 48-h period. Earlier (group 1) and later (group 2) buck-marked goats received one single insemination 12-24h or 0-12h after marking, respectively. Unmarked goats were inseminated along with group 2. In experiment 2 (n=344), FGA-treated goats were inseminated 52 and 70 h (52 h:70 h group) or 52 and 75 h (52 h:75 h group) after joining. In experiment 3 (n=285), FGA-treated goats were inseminated 52 h (1-AI group) or 52 and 75 h (2-AI group) after joining. In all experiments, an external control group given the "classical" insemination program was analysed. Over the 5-year period, 92% of the goats exhibited an LH surge during days 1-4 after joining and 98% of them ovulated. Eighty-seven percent of the LH surges detected in milk occurred during the 33-57 h interval after joining, indicating that ovulation took place around 45-69 h. In experiment 1, 96% of the goats were marked 22-70 h after joining. Kidding rate (KR; 78%) was similar between insemination groups and between FGA- and CIDR-treated goats (p>0.05). Most of the goats (95%) were inseminated during the interval between 15h before and up to 4h after ovulation. KR was not affected by the time between detection of marking and insemination or between insemination and ovulation (p>0.05). In experiment 2, KR (75%) was similar in both insemination groups (p>0.05). In experiment 3, KR was higher (p<0.05) in the 1-AI (71%) than the 2-AI group (57%). In all experiments, KR of the control group (68-73%) was similar to that achieved in goats induced to ovulate by the male effect. Prolificity (2.1+/-0.7) was not affected by any of the factors examined (p>0.05). In conclusion, high fertility can be achieved during anoestrus when 1 or 2 inseminations are performed over a 24h period, determined by oestrus or by the introduction of the buck, if light-treated goats receive 11-day FGA or CIDR treatment and are then induced to ovulate by the male effect.