Neuroanatomical organization of gonadotropin-releasing hormone neurons during the oestrus cycle in the ewe

BACKGROUND: During the preovulatory surge of gonadotropin-releasing hormone (GnRH), a very large amount of the peptide is released in the hypothalamo-hypophyseal portal blood for 24-36H00. To study whether this release is linked to a modification of the morphological organization of the GnRH-containing neurons, i.e. morphological plasticity, we conducted experiments in intact ewes at 4 different times of the oestrous cycle (before the expected LH surge, during the LH surge, and on day 8 and day 15 of the subsequent luteal phase). The cycle stage was verified by determination of progesterone and LH concentrations in the peripheral blood samples collected prior to euthanasia. RESULTS: The distribution of GnRH-containing neurons throughout the preoptic area around the vascular organ of the lamina terminalis was studied following visualisation using immunohistochemistry. No difference was observed in the staining intensity for GnRH between the different groups. Clusters of GnRH-containing neurons (defined as 2 or more neurons being observed in close contact) were more numerous during the late follicular phase (43 ± 7) than during the luteal phase (25 ± 6), and the percentage of clusters was higher during the beginning of the follicular phase than during the luteal phase. There was no difference in the number of labelled neurons in each group. CONCLUSIONS: These results indicate that the morphological organization of the GnRH-containing neurons in ewes is modified during the follicular phase. This transitory re-organization may contribute to the putative synchronization of these neurons during the surge. The molecular signal inducing this plasticity has not yet been identified, but oestradiol might play an important role, since in sheep it is the only signal which initiates the GnRH preovulatory surge

Endogenous circannual rhythm in LH secretion: insight from signal analysis coupled with mathematical modelling

In sheep as in many vertebrates, the seasonal pattern of reproduction is timed by the annual photoperiodic cycle, characterized by seasonal changes in the daylength. The photoperiodic information is translated into a circadian profile of melatonin secretion. After multiple neuronal relays (within the hypothalamus), melatonin impacts GnRH (gonadotrophin releasing hormone) secretion that in turn controls ovarian cyclicity. The pattern of GnRH secretion is mirrored into that of LH (luteinizing hormone) secretion, whose plasmatic level can be easily measured. We addressed the question of whether there exists an endogenous circannual rhythm in a tropical sheep population that exhibits clear seasonal ovarian activity when ewes are subjected to temperate latitudes. We based our analysis on LH time series collected in the course of 3 years from ewes subjected to a constant photoperiodic regime. Due to intra- and inter- animal variability and unequal sampling times, the existence of an endogenous rhythm is not straightforward. We have used time-frequency signal processing methods to extract hidden rhythms from the data. To further investigate the LF (low frequency) and HF (high frequency) components of the signals, we have designed a mathematical model of LH plasmatic level accounting for the effect of experimental sampling times. The model enables us to confirm the existence of an endogenous circannual rhythm, to investigate the action mechanism of photoperiod on the pulsatile pattern of LH secretion (control of the interpulse interval) and to conclude that the HF component is mainly due to the experimental sampling protocol

Récepteurs de la mélatonine (pharmacologie du récepteur ovin MT2, identification de leur activité constitutive et développement d'une approche par ARN interférent.)

La mélatonine est une hormone synthétisée et sécrétée uniquement la nuit par la glande pinéale. Son rôle principal est son implication dans la synchronisation de la saison de reproduction. La mélatonine se lie aux récepteurs, MT1 et MT,, membres de la famille des récepteurs à sept domaines transmembranaires couplés aux protéines G (RCPG).Le clonage récent du récepteur ovin MT2a remis en cause toutes les données connues. La pharmacologie et les voies de signalisation du récepteur oMT2ont été étudiées et sont communes à celles des récepteurs des autres espèces. En revanche, oMT2possède une originalité de séquence avec la présence du motif DRY, fortement impliqué dans l établissement de l activité constitutive des RCPG. D une part, nous avons montré que l ensemble des récepteurs MT possèdent une activité constitutive. D autre part, nous avons identifié deux agonistes inverses pour les récepteurs hMT2, initialement décrits comme antagonistes. Dans l optique de discriminer les rôles respectifs des récepteurs MT in vivo, le développement d une approche par ARN interférents a été validée dans un modèle cellulaire, la lignée CHO-KI exprimant les récepteurs ovins et de rat.Melatonin is a hormone synthesized and secreted only during night by pineal gland. A main role of melatoninconcerns its implication in the synchronization of reproductive seasonality. Binding sites of melatonin are MT1and MT2 receptors which belong to the superfamily of seven-transmembrane-spanning G protein-coupledreceptors (GPCRs).Recent cloning of ovine MT2 receptor has challenged the knowledge about melatonin receptors. Wedemonstrated that its pharmacology and signalling pathways were similar to subtype 2 receptor of othersspecies (human and rat). Nevertheless, oMT2 receptor possesses a particularity of sequence, with the presenceof DRY motif which is known to be involved in the establishment of constitutive activity of GPCRs. In ourstudy, we demonstrated the existence of constitutive activity for ail the melatonin receptors. In addition, weidentified two inverse agonists for human MT2 receptors, previously described as antagonists. To describe therespective roles of each subtype of melatonin receptors in vivo, siRNA approach was developed in cell line,CI-10-K Iexpressing ovine and rat melatonin receptorsTOURS-Bibl.électronique (372610011) / SudocSudocFranceF

Activité anti-androgénique de Leptadenia hastata (Pers.) Decne : effet compétitif des extraits aqueux de la plante et du propionate de testostérone sur des rats impubères castrés

Anti-androgenic activity of Leptadenia hastata (Pers.) Decne: competitive effect of the aqueous extracts of the plant and the testosterone propionate on castrated immature rats. The anti-androgenic activity and the evaluation of competitiveness between the extracts of Leptadenia hastata and the testosterone propionate (TP) were studied on Wistar immature castrated rats. The first group received only 0.04; 0.4; 4; 40; 400 and 1,000 µg.kg-1 of TP and the second group received simultaneously these different doses of TP and 200 mg.kg-1 of L. hastata. The various treatments showed a significant increase (p < 0.05) of the weight of androgeno-dependent organs and the level of plasmatic testosterone. At low dosis of TP, the dosis of 200 mg.kg-1 of L. hastata inhibited TP effects, whereas at high doses of TP L. hastata extracts potentiated TP effects. In conclusion, the anti-androgenic effect of the extract of L. hastata is expressed when the TP amounts are weak

Role of cerebrospinal fluid in carrying melatonin to its central target sites

Through the duration of its nocturnal secretion, melatonin is the primary transducer of photoperiodic information to the reproductive axis in seasonal breeders. In this review, we describe recent results supporting the hypothesis that melatonin, released by the pineal gland, reaches its central targets carried by cerebrospinal fluid (CSF), rather than blood. Firstly, melatonin is released in the CSF through a specific site in the pineal recess where protruding pinealocytes contact with the CSF. Consequently, melatonin is present in much higher concentrations (100 times), in the cerebrospinal fluid (CSF) than in the blood. Secondly, CSF melatonin diffuses in the brain tissues quickly and efficiently and thus reaches periventricular structures featuring 2-iodomelatonin binding sites. These data support the idea that CSF melatonin acts as a physiological signal for the brainLa mélatonine, par sa durée nocturne de sécrétion, traduit les effets de la photopériode sur la reproduction. Dans cette revue, nous décrivons des résultats récents qui étayent l'hypothèse selon laquelle la mélatonine est transportée de la glande pinéale vers ses cibles cérébrales par le liquide céphalo-rachidien (LCR), plutôt que par le sang. Premièrement, la mélatonine est libérée dans le LCR en un site spécifique dans le recessus pinéalien où des pinéalocytes sont en contact direct avec le LCR. En conséquence, la mélatonine est présente à des concentrations environ 100 fois plus élevées dans le LCR que dans le sang. Deuxièmement, la mélatonine du LCR est capable de diffuser dans les tissus cérébraux rapidement et efficacement et peut donc atteindre des structures périventriculaires dans lesquelles des sites de liaison à la 2-iodomélatonine sont présents. L'ensemble de ces données conforte l'idée que la mélatonine du LCR est un signal physiologique pour le cerveau

Genetic parameters of spontaneous spring ovulatory activity in Mérinos d'Arles sheep

The genetic parameters of spontaneous spring ovulatory activity were investigated in the Mérinos d’Arles breed under the usual pastoral and transhumant management conditions of this breed in southeastern France. Ovulatory activity was determined by assaying the plasma progesterone concentration in two blood samples taken 8-10 days apart. The data set consisted of 1 887 ovulatory activity performance measurements in 1995, 1996 and 1997 on 933 ewes, daughters of 176 rams. The effects of the ’physiological status’ (hoggets, adult ewes with or without lambing in the previous autumn), age and live weight just before the mating period were found to be highly significant. They were included in the linear animal model and the threshold sire model used to estimate genetic parameters. On average, 27.9 % of ewes exhibited ovulatory activity in April. Age and live weight just before the mating period had a marked positive effect on ovulatory activity. A difference of about 8-9 % was observed between extreme classes for these factors. The heritability and repeatability estimated through the linear model were 0.20 (standard error: 0.04) and 0.30 (0.07), respectively. When using the threshold model, the heritability was 0.37. These values led us to conclude that a genetic approach for improving spontaneous spring ovulatory activity should be further developed. Nevertheless, further studies are necessary to determine all the implications of such selection.Les paramètres génétiques de l’activité ovulatoire spontanée au printemps ont été estimés en race Mérinos d’Arles dans le système d’élevage pastoral traditionnel (transhumance estivale) du sud-est de la France. Le dosage de la progestérone plasmatique dans deux prélèvements sanguins effectués à 8-10 j d’intervalle a permis de déterminer l’activité ovulatoire des brebis. 1887 performances d’activité ovulatoire ont été enregistrées en 1995, 1996 et 1997, sur 933 brebis issues de 176 béliers. Le «statut physiologique » (antenaises, brebis adultes avec ou sans mise bas à l’automne précédent), l’âge et le poids au moment de la lutte des brebis ont des effets très significatifs sur l’activité ovulatoire. Ils ont été pris en compte dans le modèle animal linéaire et le modèle père à seuil utilisés pour estimer les paramètres génétiques. En moyenne, 27,9 % des brebis présentaient une activité ovulatoire en avril. L’âge et le poids au moment de la lutte ont un net effet positif sur l’activité ovulatoire. Une différence de 8-9 % a été observée entre les classes extrêmes pour ces facteurs. L’héritabilité et la répétabilité estimées avec le modèle linéaire sont de 0,20 (erreur standard : 0,04) et de 0,30 (0,07), respectivement. L’héritabilité calculée avec le modèle à seuil est de 0,37. En conclusion, compte tenu de ces valeurs, l’approche génétique visant à améliorer l’activité ovulatoire spontanée au printemps mérite d’être poursuivie. Néanmoins, d’autres études sont nécessaires pour connaître toutes les implications que supposent une telle sélection

Changes in the 5-HT2(A )receptor system in the pre-mammillary hypothalamus of the ewe are related to regulation of LH pulsatile secretion by an endogenous circannual rhythm

BACKGROUND: We wanted to determine if changes in the expression of serotonin 2A receptor (5HT2(A )receptor) gene in the premammillary hypothalamus are associated with changes in reproductive neuroendocrine status. Thus, we compared 2 groups of ovariectomized-estradiol-treated ewes that expressed high vs low LH pulsatility in two different paradigms (2 groups per paradigm): (a) refractoriness (low LH secretion) or not (high LH secretion) to short days in pineal-intact Ile-de-France ewes (RSD) and (b) endogenous circannual rhythm (ECR) in free-running pinealectomized Suffolk ewes in the active or inactive stage of their reproductive rhythm. RESULTS: In RSD ewes, density of 5HT2(A )receptor mRNA (by in situ hybridization) was significantly higher in the high LH group (25.3 ± 1.4 vs 21.4 ± 1.5 grains/neuron, P < 0.05) and (3)H-Ketanserin binding (a specific radioligand) of the median part of the premammillary hypothalamus tended to be higher in the high group (29.1 ± 4.0 vs 24.6 ± 4.2 fmol/mg tissu-equivalent; P < 0.10). In ECR ewes, density of 5HT2(A )receptor mRNA and (3)H-Ketanserin binding were both significantly higher in the high LH group (20.8 ± 1.6 vs 17.0 ± 1.5 grains/neuron, P < 0.01, and 19.7 ± 5.0 vs 7.4 ± 3.4 fmol/mg tissu-equivalent; P < 0.05, respectively). CONCLUSIONS: We conclude that these higher 5HT2(A )receptor gene expression and binding activity of 5HT2(A )receptor in the premammillary hypothalamus are associated with stimulation of LH pulsatility expressed before the development of refractoriness to short days and prior to the decline of reproductive neuroendocrine activity during expression of the endogenous circannual rhythm

GPR50 is the mammalian ortholog of Mel1c: Evidence of rapid evolution in mammals

<p>Abstract</p> <p>Background</p> <p>The melatonin receptor subfamily contains three members Mel1a, Mel1b and Mel1c, found in all vertebrates except for Mel1c which is found only in fish, Xenopus species and the chicken. Another receptor, the melatonin related receptor known as GPR50, found exclusively in mammals and later identified as a member of the melatonin receptor subfamily because of its identity to the three melatonin receptors despite its absence of affinity for melatonin. The aim of this study was to describe the evolutionary relationships between GPR50 and the three other members of the melatonin receptor subfamily.</p> <p>Results</p> <p>Using an <it>in silico </it>approach, we demonstrated that GPR50 is the ortholog of the high affinity Mel1c receptor. It was necessary to also study the synteny of this gene to reach this conclusion because classical mathematical models that estimate orthology and build phylogenetic trees were not sufficient. The receptor has been deeply remodelled through evolution by the mutation of numerous amino acids and by the addition of a long C-terminal tail. These alterations have modified its affinity for melatonin and probably affected its interactions with the other two known melatonin receptors MT1 and MT2 that are encoded by Mel1a and Mel1b genes respectively. Evolutionary studies provided evidence that the GPR50 group evolved under different selective pressure as compared to the orthologous groups Me11 a, b, and c.</p> <p>Conclusion</p> <p>This study demonstrated that there are only three members in the melatonin receptor subfamily with one of them (Me11c) undergoing rapid evolution from fishes and birds to mammals. Further studies are necessary to investigate the physiological roles of this receptor.</p

Genetic variability of the pattern of night melatonin blood levels in relation to coat changes development in rabbits

To assess the genetic variability in both the nocturnal increase pattern of melatonin concentration and photoresponsiveness in coat changes, an experiment on 422 Rex rabbits (from 23 males) raised under a constant light programme from birth was performed. The animals were sampled at 12 weeks of age, according to 4 periods over a year. Blood samples were taken 7 times during the dark phase and up to 1 h after the lighting began. Maturity of the fur was assessed at pelting. Heritability estimates of blood melatonin concentration (0.42, 0.17 and 0.11 at mid-night, 13 and 15 h after lights-out respectively) and strong genetic correlations between fur maturity and melatonin levels at the end of the dark phase (-0.64) indicates that (i) the variability of the nocturnal pattern of melatonin levels is under genetic control and (ii) the duration of the nocturnal melatonin increase is a genetic component of photoresponsiveness in coat changes