Morphological and immunohistochemical study of the rabbit vomeronasal organ
Authors: Villamayor Paula R. , Cifuentes Jose Manuel , Fdz.‐de‐Troconiz Patricia , Sanchez‐Quinteiro Pablo 
Published: September 2018

The characterization of the rabbit mammary pheromone, which is sensed by the main olfactory system, has made this species a unique model for the study of pheromonal communication in mammals. This discovery has brought attention to the global understanding of chemosensory communication in this species. Chemocommunication is mediated by two distinct organs located in the nasal cavity, the main olfactory epithelium and the vomeronasal organ (VNO). However, there is a lack of knowledge about the vomeronasal system in rabbits. To understand the role of this system, an exhaustive anatomical and histological study of the rabbit VNO was performed. The rabbit VNO was studied macroscopically by light microscopy, and by histochemical and immunohistochemical techniques. We employed specific histological staining techniques (periodic acid‐Schiff, Alcian blue, Gallego's trichrome), confocal autofluorescence, histochemical labelling with the lectin Ulex europaeus agglutinin (UEA‐I), and immunohistochemical studies of the expression of the Gαi2 and Gαo proteins and olfactory marker protein. The opening of the vomeronasal duct into the nasal cavity and its indirect communication with the oral cavity through a functional nasopalatine duct was demonstrated by classical dissection and microdissection. In a series of transverse histological sections, special attention was paid to the general distribution of the various soft‐tissue components of this organ (duct, glands, connective tissue, blood vessels and nerves) and to the nature of the capsule of the organ. Among the main morphological features that distinguish the rabbit VNO, the presence of a double envelope, which is bony externally and cartilaginous internally, and highly developed venous sinuses stand out. This observation indicates the crucial role played in this species by the pumping mechanism that introduces chemical signals into the vomeronasal duct. The functional properties of the organ are also confirmed by the presence of a well‐developed neuroepithelium and profuse glandular tissue that is positive for neutral mucopolysaccharides. The role of glycoconjugates was assessed by the identification of the α1‐2 fucose glycan system in the neuroepithelium of the VNO employing UEA‐I lectin. The pattern of labelling, which was concentrated around the commissures of the sensory epithelium and more diffuse in the central segments, is different from that found in most mammals studied. According to the expression of G‐proteins, two pathways have been described in the VNOs of mammals: neuroreceptor cells expressing the Gαi2 protein (associated with vomeronasal receptor type 1); and cells expressing Gαo (associated with vomeronasal receptor type 2). The latter pathway is absent in most mammals studied. The expression of both G‐protein families in the rabbit VNO places Lagomorpha together with rodents and insectivores in a small group of mammals belonging to the two‐path model. These findings support the notion that the rabbit possesses a highly developed VNO, with many specific morphological features, which highlights the significance of chemocommunication in this species.