Vomeronasal Research Articles

Does the Environmental Air Impact the Condition of the Vomeronasal Organ? A Mouse Model for Intensive Farming.

Chemical communication in mammals is ensured by exchanging chemical signals through the vomeronasal organ (VNO) and its ability to detect pheromones. The alteration of this organ has been proven to impact animal life, participating in the onset of aggressive behaviors in social groups. To date, few studies have highlighted the possible causes leading to these alterations, and the farming environment has not been investigated, even though irritant substances such as ammonia are known to induce serious damage in the respiratory tract. The goal of this study was to investigate the environmental impact on the VNO structure. Thirty mice were split into three groups, one housed in normal laboratory conditions and the other two in confined environments, with or without the release of litter ammonia. VNOs were analyzed using histology and immunohistochemistry to evaluate the effect of different environments on their condition. Both restricted conditions induced VNO alterations (p = 0.0311), soft-tissue alteration (p = 0.0480), and nonsensory epithelium inflammation (p = 0.0024). There was glycogen accumulation (p < 0.0001), the olfactory marker protein was underexpressed (p < 0.0001), and Gαi2 positivity remained unchanged while Gαo expression was upregulated in confined conditions. VNO conditions seemed to worsen with ammonia, even if not always significantly. These murine model results suggest that the housing environment can strongly impact VNO conditions, providing novel insights for improving indoor farming systems.

Natural and Pathological Aging Distinctively Impacts the Pheromone Detection System and Social Behavior

Normal aging and many age-related disorders such as Alzheimer’s disease cause deficits in olfaction; however, it is currently unknown how natural and pathological aging impacts the detection of social odors which might contribute to the impoverishment of social behavior at old age further worsening overall health. Analysis of the vomeronasal organ, the main gateway to pheromone-encoded information, indicated that natural and pathological aging distinctively affects the neurogenic ability of the vomeronasal sensory epithelium. Whereas cell proliferation remained majorly preserved in 1-year-old APP/PS1 mice, naturally aged animals exhibited significant deficiencies in the number of mature, proliferative, and progenitor cells. These alterations may support age-related deficits in the recognition of social cues and the display of social behavior. Our findings indicate that aging disrupts the processing of social olfactory cues decreasing social odor exploration, discrimination, and habituation in both wild-type senescent (2-year-old) mice and in 1-year-old double mutant model of Alzheimer’s disease (APP/PS1). Furthermore, social novelty was diminished in 1-year-old APP/PS1 mice, indicating that alterations in the processing of social cues are accelerated during pathological aging. This study reveals fundamental differences in the cellular processes by which natural and pathological aging disrupts the exploration of social information and social behavior.

Parallel Olfactory Systems Synergistically Activate the Posteroventral Part of the Medial Amygdala Upon Alarm Pheromone Detection in Rats

In rats, a mixture of hexanal and 4-methylpentanal is a main component of the alarm pheromone. When detected by the main olfactory system (MOS) and the vomeronasal system, respectively, they activate the anterior part of the bed nucleus of the stria terminalis (BNSTa). Therefore, the information from the two olfactory systems is expected to be integrated before being transmitted to the BNSTa. To specify the integration site, we examined Fos expression in 16 brain regions in response to water (n = 10), hexanal (n = 9), 4-methylpentanal (n = 9), the mixture (n = 9), or the alarm pheromone (n = 9) in male rats. The posteroventral part of the medial amygdala showed increased Fos expression to hexanal and 4-methylpentanal. The expression was further increased by the mixture. Therefore, this region is suggested as the integration site. In addition, the BNSTa, paraventricular nucleus of the hypothalamus, and anteroventral, anterodorsal, and posterodorsal parts of the medial amygdala were suggested to be located downstream of the integrated site because only the mixture increased Fos expression. We suggest that the posterolateral part of the cortical amygdala is upstream of the integration site in the MOS because all stimuli increased Fos expression. The posterior part of the bed nucleus of the stria terminalis and posteromedial part of the cortical amygdala were suggested as being located upstream in the vomeronasal system because 4-methylpentanal and the mixture increased Fos expression. These results provide information about the neural pathway underlying the alarm pheromone effects.

Transient Receptor Potential channels (TRP) in GtoPdb v.2023.1

Transient Receptor Potential channels (TRP) in GtoPdb v.2023.1

Spatiotemporal dynamics of the development of mouse olfactory system from prenatal to postnatal period.

The olfactory epithelium (OE) and olfactory bulb (OB) are the major components of the olfactory system and play critical roles in olfactory perception. However, the embryonic development of OE and OB by using the olfactory specific genes has not been comprehensively investigated yet. Most previous studies were limited to a specific embryonic stage, and very little is known, till date, about the development of OE. The current study aimed to explore the development of mouse olfactory system by spatiotemporal analysis of the histological features by using the olfactory specific genes of olfactory system from the prenatal to postnatal period. We found that OE is divided into endo-turbinate, ecto-turbinate, and vomeronasal organs, and that putative OB with putative main and accessory OB is formed in the early developmental stage. The OE and OB became multilayered in the later developmental stages, accompanied by the differentiation of olfactory neurons. Remarkably, we found the development of layers of olfactory cilia and differentiation of OE to progress dramatically after birth, suggesting that the exposure to air may facilitate the final development of OE. Overall, the present study laid the groundwork for a better understanding of the spatial and temporal developmental events of the olfactory system.

Bilateral ablation of the vomeronasal organs produces dramatic changes in the EEG of the main olfactory system during paradoxical sleep

Background: The vomeronasal organ (VNO) plays an important role in reproductive physiology, behavior, sexual behavior and aggression. Objective: The aim is to report a surprising effect of the removal of the VNO on the cortical electrical activity of the olfactory bulb, piriform lobe and frontal cortex exclusively during paradoxical sleep (PS). Methodology: Seventeen adult armadillos chronically prepared for electrographic recordings were employed. Some animals were subjected to VNO bilateral remove. They were studied during wakefulness and sleep phases. Results: An outstanding phenomenon occurs during PS of armadillos submitted to bilateral ablation of the VNO. There is a dramatic change in the oscillations of the olfactory bulb electrical activity strictly confined to PS. The change consists of the sudden appearance, as soon as PS initiates, of high amplitude gamma oscillations during the whole duration of each PS episode. They disappear as soon as PS comes to an end. The oscillations are also seen in the paleocortex and the frontal neocortex. They disappear after the transverse section of the olfactory peduncles. Conclusions: Our results show the absence of VNO deeply modifies the electrophysiological expression of PS in the olfactory bulb and other extensive brain areas. They are elicited by the centrifugal input to the olfactory bulb. From there they propagate to the piriform and frontal neocortex. These results show a new and subtle role for the organs. Some hypotheses related to the effects of bilateral removal of the VNO are presented.

Bilateral ablation of the vomeronasal organs produces dramatic changes in the EEG of the main olfactory system during paradoxical sleep

Bilateral ablation of the vomeronasal organs produces dramatic changes in the EEG of the main olfactory system during paradoxical sleep

Type 1 vomeronasal receptor expression in juvenile and adult lungfish olfactory organ

Lungfish are the most closely related fish to tetrapods. The olfactory organ of lungfish contains lamellae and abundant recesses at the base of lamellae. Based on the ultrastructural and histochemical characteristics, the lamellar olfactory epithelium (OE), covering the surface of lamellae, and the recess epithelium, contained in the recesses, are thought to correspond to the OE of teleosts and the vomeronasal organ (VNO) of tetrapods. With increasing body size, the recesses increase in number and distribution range in the olfactory organ. In tetrapods, the expression of olfactory receptors is different between the OE and VNO; for instance, the type 1 vomeronasal receptor (V1R) is expressed only in the OE in amphibians and mainly in the VNO in mammals. We recently reported that V1R-expressing cells are contained mainly in the lamellar OE but also rarely in the recess epithelium in the olfactory organ of lungfish of approximately 30 cm body length. However, it is unclear whether the distribution of V1R-expressing cells in the olfactory organ varies during development. In this study, we compared the expression of V1Rs in the olfactory organs between juveniles and adults of the African lungfish Protopterus aethiopicus and South American lungfish, Lepidosiren paradoxa. The density of V1R-expressing cells was higher in the lamellae than in the recesses in all specimens evaluated, and this pattern was more pronounced in juveniles than adults. In addition, the juveniles showed a higher density of V1R-expressing cells in the lamellae compared with the adults. Our results imply that differences in lifestyle between juveniles and adults are related to differences in the density of V1R-expressing cells in the lamellae of lungfish.

Contactin-6-deficient male mice exhibit the abnormal function of the accessory olfactory system and impaired reproductive behavior.

Contactin-6 (CNTN6), also known as NB-3, is a neural recognition molecule and a member of the contactin subgroup of the immunoglobulin superfamily. Gene encoding CNTN6 is expressed in many regions of the neural system, including the accessory olfactory bulb (AOB) in mice. We aim to determine the effect of CNTN6 deficiency on the function of the accessory olfactory system (AOS). We examined the effect of CNTN6 deficiency on the reproductive behavior of male mice through behavioral experiments such as urine sniffing and mate preference tests. Staining and electron microscopy were used to observe the gross structure and the circuitry activity of the AOS. Cntn6 is highly expressed in the vomeronasal organ (VNO) and the AOB, and sparsely expressed in the medial amygdala (MeA) and the medial preoptic area (MPOA), which receive direct and/or indirect projections from the AOB. Behavioral tests to examine reproductive function in mice, which is mostly controlled by the AOS, revealed that Cntn6-/- adult male mice showed less interest and reduced mating attempts toward estrous female mice in comparison with their Cntn6+/+ littermates. Although Cntn6-/- adult male mice displayed no obvious changes in the gross structure of the VNO or AOB, we observed the increased activation of granule cells in the AOB and the lower activation of neurons in the MeA and the MPOA as compared with Cntn6+/+ adult male mice. Moreover, there were an increased number of synapses between mitral cells and granule cells in the AOB of Cntn6-/- adult male mice as compared with wild-type controls. These results indicate that CNTN6 deficiency affects the reproductive behavior of male mice, suggesting that CNTN6 participated in normal function of the AOS and its ablation was involved in synapse formation between mitral and granule cells in the AOB, rather than affecting the gross structure of the AOS.

The clinical significance of the human vomeronasal organ

ObjectiveTo find out whether the vomeronasal organ (VNO) can be identified in the nose as a mucosal pit in the anterior nasal septum, to elucidate its function in man and to determine whether it is important to preserve the VNO during septal surgery.MethodsLiterature review.Results and conclusionThe VNO is histologically present in almost all humans, but a macroscopically visible septal pit does not necessarily correspond with the actual VNO. The human VNO is probably a vestigial organ with a non-operational sensory function. It is not necessary to take particular care not to damage the VNO during septal surgery.

Behavioural disorder in a dog with congenital agenesis of the vomeronasal organ and the septum pellucidum

ABSTRACTA 1.5‐year‐old, entire male mongrel dog with a bifid nose and primary cleft palate showed aggressiveness towards unknown dogs despite having adequate canine socialisation and the absence of previous traumatic experiences; furthermore, he does not exhibit sexual behaviour towards females in heat. He was previously treated for fear‐related aggression towards unknown people and environmental phobia due to poor human socialisation. Pharmacological treatment with fluoxetine and trazodone, together with behaviour modification, was started without success. Therefore, it was decided to repeat medical tests. CT and MRI revealed a rostral middle raphe ossification defect incompatible with the presence of the vomeronasal organ (VNO) and the absence of the septum pellucidum (SP). The dog was presumptively diagnosed with aggressiveness towards unknown dogs due to an organic cause, a congenital anomaly with agenesis of the VNO and SP. This case shows the importance of both structures in the regular intraspecific interactions of dogs.

Atavistic and vestigial anatomical structures in the head, neck, and spine: an overview.

Organisms may retain nonfunctional anatomical features as a consequence of evolutionary natural selection. Resultant atavistic and vestigial anatomical structures have long been a source of perplexity. Atavism is when an ancestral trait reappears after loss through an evolutionary change in previous generations, whereas vestigial structures are remnants that are largely or entirely functionless relative to their original roles. While physicians are cognizant of their existence, atavistic and vestigial structures are rarely emphasized in anatomical curricula and can, therefore, be puzzling when discovered incidentally.In addition, the literature is replete with examples of the terms atavistic and vestigial being used interchangeably without careful distinction between them. We provide an overview of important atavistic and vestigial structures in the head, neck, and spine that can serve as a reference for anatomists and clinical neuroscientists. We review the literature on atavistic and vestigial anatomical structures of the head, neck, and spine that may be encountered in clinical practice. We define atavistic and vestigial structures and employ these definitions consistently when classifying anatomical structures. Pertinent anatomical structures are numerous and include human tails, plica semilunaris, the vomeronasal organ, levator claviculae, and external ear muscles, to name a few. Atavistic and vestigial structures are found throughout the head, neck, and spine. Some, such as human tails and branchial cysts may be clinically symptomatic. Literature reports indicate that their prevalence varies across populations. Knowledge of atavistic and vestigial anatomical structures can inform diagnoses, prevent misrecognition of variation for pathology, and guide clinical interventions.

Flehmen, Osteophagia, and Other Behaviors of Giraffes (Giraffa giraffa angolensis): Vomeronasal Organ Adaptation.

The size of adult male giraffes (Giraffa giraffa angolensis) far exceeds the size of the females. At the Namutoni waterholes in Etosha National Park, bulls were seen many times each day screening adult females for their pending sexual receptivity by provoking them to urinate; this mainly involved sniffing their genitalia. If the female accedes to the male's invitation, she widens her hindleg stance, braces her body, and then urinates, usually for at least five seconds. The male places his muzzle and tongue in the urine stream, and then performs flehmen, often raising his head high in the air. Males never investigated urine on the ground. The bilateral papillae on the giraffe's hard palate connect with the nasopalatine ducts, which enter the bilateral vomeronasal organ (VNO). Unlike many mammals, the giraffe's VNO lacks a prominent connection to the nasal cavity and its connections to the oral cavity are primarily via the incisive papillae. Most days, some giraffes were observed searching for bones for extended periods, chewing them, and sometimes being troubled by a bone stuck in their mouth. A giraffe killed by lions was frequented for several days by a procession of giraffes investigating the carcass. A very dark-colored bull giraffe emitted loud pulsed growls that drove off most of the surrounding giraffes.

The olfactory limbus of the red fox (Vulpes vulpes). New insights regarding a noncanonical olfactory bulb pathway.

Introduction: The olfactory system in most mammals is divided into several subsystems based on the anatomical locations of the neuroreceptor cells involved and the receptor families that are expressed. In addition to the main olfactory system and the vomeronasal system, a range of olfactory subsystems converge onto the transition zone located between the main olfactory bulb (MOB) and the accessory olfactory bulb (AOB), which has been termed the olfactory limbus (OL). The OL contains specialized glomeruli that receive noncanonical sensory afferences and which interact with the MOB and AOB. Little is known regarding the olfactory subsystems of mammals other than laboratory rodents. Methods: We have focused on characterizing the OL in the red fox by performing general and specific histological stainings on serial sections, using both single and double immunohistochemical and lectin-histochemical labeling techniques. Results: As a result, we have been able to determine that the OL of the red fox (Vulpes vulpes) displays an uncommonly high degree of development and complexity. Discussion: This makes this species a novel mammalian model, the study of which could improve our understanding of the noncanonical pathways involved in the processing of chemosensory cues.

Glycoconjugate-Specific Developmental Changes in the Horse Vomeronasal Organ

The vomeronasal organ (VNO) is a tubular pheromone-sensing organ in which the lumen is covered with sensory and non-sensory epithelia. This study used immunohistochemistry and lectin histochemistry techniques to evaluate developmental changes, specifically of the glycoconjugate profile, in the horse VNO epithelium. Immunostaining analysis revealed PGP9.5 expression in some vomeronasal non-sensory epithelium (VNSE) cells and in the vomeronasal receptor cells of the vomeronasal sensory epithelium (VSE) in fetuses, young foals, and adult horses. Olfactory marker protein expression was exclusively localized in receptor cells of the VSE in fetuses, young foals, and adult horses and absent in VNSE. To identify the glycoconjugate type, lectin histochemistry was performed using 21 lectins. Semi-quantitative analysis revealed that the intensities of glycoconjugates labeled with WGA, DSL, LEL, and RCA₁₂₀ were significantly higher in adult horse VSE than those in foal VSE, whereas the intensities of glycoconjugates labeled with LCA and PSA were significantly lower in adult horse VSE. The intensities of glycoconjugates labeled with s-WGA, WGA, BSL-II, DSL, LEL, STL, ConA, LCA, PSA, DBA, SBA, SJA, RCA₁₂₀, jacalin, and ECL were significantly higher in adult horse VNSE than those in foal VNSE, whereas the intensity of glycoconjugates labeled with UEA-I was lower in adult horse VNSE. Histochemical analysis of each lectin revealed that various glycoconjugates in the VSE were present in the receptor, supporting, and basal cells of foals and adult horses. A similar pattern of lectin histochemistry was also observed in the VNSE of foals and adult horses. In conclusion, these results suggest that there is an increase in the level of N-acetylglucosamine (labeled by WGA, DSL, LEL) and galactose (labeled by RCA₁₂₀) in horse VSE during postnatal development, implying that they may influence the function of VNO in adult horses.

Микроморфологическая характеристика рецепторной части дополнительной обонятельной системы бобра обыкновенного

The article presents the microscopic organization of the vomeronasal organ (VNO), membranes, vascular system and innervation of the VNO of the common beaver. A comparative analysis of the patterns and features of the structure of the organ in comparison with other representatives of the mammalian family is given. The research was carried out on the basis of the Department of Anatomy and Histology of Animals named after Professor A. F. Klimov of the Moscow State Academy of Veterinary Medicine and Biotechnology – MBA named after K. I. Scriabin. The objects of research were 10 common beavers aged 2 to 6 years, without pronounced signs of pathology of the nasal and oral cavities. The animals were selected from hunting farms of the Moscow region in connection with planned slaughter. When studying the macro- and micromorphological features of the vomeronasal organ, a comprehensive methodological approach was used, including classical and fine anatomical dissection under the control of a binocular magnifying glass with illumination, followed by a description of the studied structures, as well as histological examination with further light microscopy of sections of VNO stained with hematoxylin and eosin and toluidine blue by the Nissl method. It was found that the common beaver has a vomeronasal organ, which is a receptor part of the additional olfactory system, has general patterns of morphological organization characteristic of representatives of other taxonomic categories. At the same time, we have identified species-specific micromorphological features characteristic of representatives of the rodent family. So in the middle part of the ventral wall of the cavity of the external cavity there is a specific depression, the multilayer epithelium of which differs from the border epithelium on both sides. In addition, lacunar-type capillaries are located in the own plate of the sensory epithelium, a large muscle-type vein with a large number of somatic-type capillaries is located contralaterally. The endings of the vomeronasal nerve (VNN) in the walls of the ordinary beaver have their own topographic features. In the area of the mucous membrane of the sensory epithelium, the nerve branches are located transversely along the course of the VNO channel. The innervation of the mucous membrane of the respiratory epithelium has the opposite direction – the branches of the nerve are located along the course of the organ channel. In the submucosal base of the mucous membrane from the respiratory epithelium, ventrally from the wall of the muscle-type vein, there are single, small singly located neurons of the intramural ganglion.

Characterization of Vomeronasal Receptor Class 2 in Danio rerio

The best three distinct families of putative pheromone receptors have been identified in the vomeronasal organ (V1Rs, V2Rs and V3Rs). All are G protein-coupled receptors but are only distantly related to the receptors of the main olfactory system, highlighting their different role. Objective: To characterize the Vomeronasal receptor 2 gene family in Zebra Fish (Danio rerio). Methods: Extensive survey was done to choose top V2R genes. Different software and tools were used to characterize those genes including Eggnog 2.0, MAFFT, ITOL, Weblogo and SOSUI Signal. Results: In order to get insights into this gene family in Zebra fish, we performed an extensive survey of V2R derived datasets. We identified 62 genes distributed among Danio rerio encoding putative vomeronasal proteins. V2R gene family was found to be highly conserved in this study by using Weblogo. It evolved at the level of eukaryotes. The V2R is involved mainly in olfaction. Conclusions: The basic repertoire of V2R genes seems to be larger for most of the species including Danio rerio and gene duplication still plays a role in lineage-specific increases in diversity. V2R gene family is very ancient, has high duplicability suggesting its essentiality.

Pheromones, binding proteins, and olfactory systems in the pig (Sus scrofa): An updated review.

Pigs utilize multimodal communication for reproductive and other behaviors, and chemical communication is one of the key components. The success of reproduction relies on chemical communication favored by the steroid pheromones from boar saliva. These steroids were proven to be involved in advancing puberty in gilts (the boar effect) and in promoting estrus behaviors in gilts/sows, thereby helping to detect estrus and facilitating the timing of artificial insemination. The steroid pheromones bound with carrier proteins are evidenced in the mandibular (submandibular) salivary secretions of the boar. These salivary steroids bind with carrier proteins in the nasal mucus and vomeronasal organ (VNO) of the sows, eventually triggering a cascade of activities at the olfactory and endocrine levels. Besides steroid pheromones, pig appeasing pheromones (from mammary skin secretions of sows) have also been demonstrated to bind with carrier proteins in the nasal mucus and VNO of the piglets. Thus far, four different proteins have been identified and confirmed in the nasal mucus and VNO of pigs, including odorant binding proteins (OBPs), salivary lipocalin (SAL), pheromaxein, and Von Ebner's Gland Protein (VEGP). The critical roles of the chemosensory systems, main olfactory systems and VNO, have been comprehensively reported for pigs. This review summarizes the current knowledge on pheromones, their receptor proteins, and the olfactory systems of porcine species.

Interpretive review: Semiochemicals in domestic pigs and dogs.

This interpretive review includes discussion of the available scientific literature with interpretations by the authors. The broad field of semiochemicals can be confusing to scientists and consumers. This review attempts to summarize the known scientific studies for pig and dog semiochemicals while at the same time attempting to refine our use of terminology. The specific objectives of this interpretive review are to summarize and interpret much of the key scientific literature (but not the lay literature) on semiochemicals in pigs and dogs to include (1) definitions of semiochemicals and related molecules including pheromones, (2) to briefly summarize olfactory organs, and (3) and to examine the scientific literature for semiochemical mechanisms and applications in dogs and pigs (two domesticated species with known olfactory acuity). Dogs and pigs have olfactory features that are similar in that they both lack certain olfactory organs (Grueneberg ganglion and Septal Organ) and they have a small vomeronasal organ (VNO) without some major receptors that are found in other species. The primary olfactory organs for both pigs and dogs are the main olfactory epithelium and perhaps the trigeminal nerve. Several examples of pheromones activating the brain via the MOE or Trigeminal nerve rather than the VNO challenge the concept that the VNO is the site of pheromone sensing. We believe it is not appropriate to label something a pheromone when evidence is not available to show that it is a pheromone. We offer definitions for the terms semiochemicals, pheromones, interomones and others and then determine if the evidence is sufficient to call certain semiochemicals a pheromone. Here we review mixed, largely negative, scientific reports of the efficacy of some products labeled as "pheromones" that are more appropriately called semiochemicals. Interomones can have a more powerful effect on dog behavior and physiology than semiochemicals marketed as pheromones. Because marketing of semiochemicals is far ahead of the science, bringing some logic and uniformity to the field will benefit animals and hopefully cause less consumer confusion. Semiochemicals have the potential to offer powerful solutions to behavioral problems using more naturally occurring molecules.

Sex separation unveils the functional plasticity of the vomeronasal organ in rabbits

Chemosensory cues are vital for social and sexual behaviours and are primarily detected and processed by the vomeronasal system (VNS), whose plastic capacity has been investigated in mice. However, studying chemosensory plasticity outside of laboratory conditions may give a more realistic picture of how the VNS adapts to a changing environment. Rabbits are a well-described model of chemocommunication since the discovery of the rabbit mammary pheromone and their vomeronasal organ (VNO) transcriptome was recently characterised, a first step to further study plasticity-mediated transcriptional changes. In this study, we assessed the plastic capacity of the rabbit male and female VNO under sex-separation vs. sex-combined scenarios, including adults and juveniles, to determine whether the rabbit VNO is plastic and, if so, whether such plasticity is already established at early stages of life. First, we characterised the number of differentially expressed genes (DEGs) between the VNO of rabbit male and female under sex-separation and compared it to sex-combined individuals, both in adults and juveniles, finding that differences between male and female were larger in a sex-separated scenario. Secondly, we analysed the number of DEGs between sex-separated and sex-combined scenarios, both in males and females. In adults, both sexes showed a high number of DEGs while in juveniles only females showed differences. Additionally, the vomeronasal receptor genes were strikingly downregulated in sex-separated adult females, whereas in juveniles upregulation was shown for the same condition, suggesting a role of VRs in puberty onset. Finally, we described the environment-modulated plastic capacity of genes involved in reproduction, immunity and VNO functional activity, including G-protein coupled receptors. Our results show that sex-separation induces sex- and stage-specific gene expression differences in the VNO of male and female rabbit, both in adults and juveniles. These results bring out for the first time the plastic capacity of the rabbit VNO, supporting its functional adaptation to specifically respond to a continuous changing environment. Finally, species-specific differences and individual variability should always be considered in VNO studies and overall chemocommunication research.