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.

Variation of ewe olfactory secretome during a ram effect.

Introduction: Under temperate latitudes, reproduction in Ovis aries displays a marked seasonality, governed by the photoperiod. In natural conditions, the transition between sexual rest and sexual activity in both sexes is induced by the decrease of day light. Meanwhile, specific odors emitted by a sexually active male are able to reactivate the gonadotropic axis of anovulatory ewes. This physiological effect is called "male effect", precisely ram effect in the ovine species. We have previously shown that the secreted proteins, namely Olfactory Binding Proteins (OBP), contained in the nasal mucus constitute the olfactory secretome (OS), the composition of which is determined by the status of oestrus cycle of females and differs between sexual rest and sexual activity periods. The objective of this study was to test the hypothesis that exposure to sexually active male can also modify the composition of ewes olfactory secretome during a male effect, as well as hormones produced by the reactivation of the oestrus cycle in sexual activity period under natural conditions. Methods: We have set up a new non-invasive protocol of nasal mucus sampling and collected it from 12 ewes at different times during a ram effect. We analyzed the composition of their olfactory secretome by proteomics, mainly SDS-PAGE and MALDI-TOF mass spectrometry. As post-translational modifications of OBPs were a hallmark of ewes' sexual activity period, we were looking for glycosylation by western-blot and mass spectrometry. Results: The efficiency of male effect was low in stimulated ewes as only 3 females displayed elevated progesterone levels in their blood. Besides, half of control ewes (non-stimulated ones) were cycled. We noticed a common OS profile in ewes in anoestrus, versus OS of cycled ones. A very clear and important result was the apparition of O-GlcNAcylation, previously detected only in sexual activity, after only 30 min of male introduction into the flock. Discussion: This exploratory study paves the way for further experiments with larger flock to confirm and reinforce these results, and for eventually exploiting the nasal mucus as an indicator of females' receptivity to male odors.