Analysis of the vomeronasal organ transcriptome reveals variable gene expression depending on age and function in rabbits.

The vomeronasal organ (VNO) is a chemosensory organ specialized in pheromone detection that shows a broad morphofunctional and genomic diversity among mammals. However, its expression patterns have only been well-characterized in mice. Here, we provide the first comprehensive RNA sequencing study of the rabbit VNO across gender and sexual maturation stages. We characterized the VNO transcriptome, updating the number and expression of the two main vomeronasal receptor families, including 128 V1Rs and 67 V2Rs. Further, we defined the expression of formyl-peptide receptor and transient receptor potential channel families, both known to have specific roles in the VNO. Several sex hormone-related pathways were consistently enriched in the VNO, highlighting the relevance of this organ in reproduction. Moreover, whereas juvenile and adult VNOs showed significant transcriptome differences, male and female did not. Overall, these results contribute to understand the genomic basis of behavioural responses mediated by the VNO in a non-rodent model.

Histochemical identification of carbohydrate moieties in the accessory olfactory bulb of the mouse using a panel of lectins.

Lectin binding patterns in the olfactory bulb of the mouse were investigated using 12 biotinylated lectins. Three, with specificities for galactose, N-acetylgalactosamine and L-fucose, stained only the nervous and glomerular layers of the accessory olfactory bulb; four, with specificities for galactose or N-acetylglucosamine, stained these layers in both the accessory and the main olfactory bulbs; three, with specificities for N-acetylgalactosamine or L-fucose, effected general staining with little contrast between the background and the accessory olfactory bulb or other structures; the remaining two, both of them specific for mannose, stained no part of the tissue studied. In the nervous and glomerular layers of the accessory olfactory bulb six lectins stained the anterior and posterior halves with different intensities and two of these six similarly differentiated between rostral and caudal regions of the posterior half. We conclude that: (i) three lectins binding to different monosaccharides are specific stains for the vomeronasal system when used in this area of the mouse brain; (ii) it may be appropriate to distinguish three parts in the mouse accessory olfactory bulb, instead of the hitherto generally accepted two.

A descriptive and comparative lectin histochemical study of the vomeronasal system in pigs and sheep.

The accessory olfactory bulb (AOB) is the primary target of the sensory epithelium of the vomeronasal organ (VNO), and thus constitutes a fundamental component of the accessory olfactory system, which is involved in responses to behaviour-related olfactory stimuli. In this study we investigated the characteristics of the AOB, VNO, vomeronasal nerves (VNNs) and caudal nasal nerve (CdNN) in pigs and sheep, species in which olfaction plays a key behavioural role both in the neonatal period and in adulthood. The patterns of staining of the AOB by the Bandeiraea simplicifolia and Lycopersicon esculentum lectins were the same in the 2 species, whereas the Ulex europeus and Dolichos biflorus lectins gave different patterns. In both species, lectin staining of the AOB was consistent with that of the VNNs, while the CdNN did not label any of the structures studied. The entire sensory epithelium of the pig was labelled by Ulex europeus and Lycopersicum esculentum lectins, and all 4 lectins used labelled the mucomicrovillar surface of the sensory epithelium in sheep.

Lectin binding patterns in the vomeronasal organ and accessory olfactory bulb of the rat.

A number of previous studies have indicated that lectin histochemistry is an obvious choice for characterizing the vomeronasal system. However, apparently inconsistent results have been obtained: notably, the affinity with which various lectins bind to the accessory olfactory bulb varies among taxa, even considering closely related species. In the present study, the binding patterns of seven lectins in the rat accessory olfactory bulb, vomeronasal nerves and vomeronasal duct were investigated. The Bandeiraea simplicifolia lectin bound exclusively to the vomeronasal nerve and glomerular layers of the accessory olfactory bulb, while the Ulex europeus and Lycopersicon esculentum lectins bound to these regions and additionally to the nerve and glomerular layers of the main olfactory bulb. Soybean agglutinin showed a similar pattern to that obtained with the Ulex europeus and Lycopersicon esculentum lectins, though it also faintly labelled other parts of the structures examined. The Vicia villosa and Erythrina cristagalli lectins were not specific for the vomeronasal system, since they labelled grey and white matters in structures including the lateral olfactory tract and the anterior olfactory nuclei. The Dolichos biflorus lectin did not bind to vomeronasal tissues. The observed patterns of binding in the accessory olfactory bulb were consistent with those observed in the vomeronasal nerves, but unlike those observed in the epithelium of the vomeronasal duct. This latter result probably reflects binding of lectins to sugar residues contained in secreted mucus rather than those in epithelial nerve endings.

Supporting tissue and vasculature of the mammalian vomeronasal organ: the rat as a model.

The blood supply and osseocartilaginous support structures of the vomeronasal organ of the rat were studied. The study focused on adults, though 3- to 18-day-old animals were also examined. The techniques used included dissection and microdissection, injection of the vascular system with Araldite or with Indian ink in agar or gelatine, conventional histology, and scanning and transmission electron microscopy. The results indicated that blood reaches the vomeronasal organ via a branch of the sphenopalatine artery, and drains into an associated vein. Within the organ, one vein stood out by virtue of its size; this vein is accompanied by lesser veins, together with arterioles, capillaries, and lymphatic vessels. Connective tissue was readily apparent, though its distribution was heterogeneous. Analysis of series of transverse sections indicates that, in adults, the capsule that encases the vomeronasal organ is bony; in younger animals, the capsule is bony externally and cartilaginous internally; in very young animals, the capsule is entirely cartilaginous. However, it was noted that the change from cartilage to bone was due not to ossification of the existing cartilage, but to physical displacement of that cartilage by an extension of the vomer and incisive bones. Taken together, these results confirm the importance of considering the morphology of the vomeronasal organ as a whole, since there are major changes from rostral to caudal ends. Secondly, our findings regarding blood supply and the nature of the capsule support the view that the vomeronasal organ acts as a pump.

Origin and regional distribution of the arterial vessels of the vomeronasal organ in the sheep. A methodological investigation with scanning electron microscopy and cutting-grinding technique.

The origin and location of the arteries of the vomeronasal organ (VNO) in the sheep were studied by means of dissection, scanning electron microscopy of corrosion casts, and the cutting-grinding technique after injection with Araldite CY23-HY2967 via one of the carotid arteries. Dissection revealed that the most ventral of the three main branches of the sphenopalatine artery is responsible for the blood supply to the VNO. Scanning electron microscopy of corrosion casts revealed that the arterioles of the vomeronasal organ form a microvascular network. Cross sections of the region of the nasal cavity containing the VNO, obtained by the cutting-grinding technique, showed that the arterioles of the vomeronasal plexus are located medial and ventral to the vomeronasal duct. These results confirm the usefulness of the cutting-grinding technique as a complementary procedure in morphological studies of structures containing hard tissues.

Distribution of the arterial supply to the vomeronasal organ in the cat.

BACKGROUND: The main goal of this work was to investigate the general distribution of arterial blood around and inside the vomeronasal organ (VNO) of the cat. METHODS: Macro- and microdissection methods together with light and scanning electron microscopy were used. Heads were injected with an India ink/agar mixture (the VNO subsequently being cut in transverse, sagittal and horizontal sections), with clear latex (the VNO subsequently being cut in transverse sections), or with an epoxy resin to obtain casts for examination by scanning electron microscopy. RESULTS: Dissection and microdissection show that the infraorbital, minor palatine, and descending palatine arteries have a common origin, rostral to the Rete mirabile arteria maxillaris. In transverse series and in the rostral half of the VNO, an arteriole is consistently observed between the vomeronasal duct and the lateral sheet of the vomeronasal cartilage. In this same segment, arterial branches with different orientations (perpendicular, horizontal, or transverse with respect to the main axis of the organ) are observed. Scanning electron microscopy of arterial casts shows that arterial vessels of the mucosa of the nasal septum have a direct relationship with the VNO. CONCLUSIONS: Branches of the sphenopalatine artery are the chief route of blood supply to the VNO. The vomeronasal parenchyma has few arterial vessels, and these are usually situated in the same position. Differences observed between the arteries inside and outside the VNO and the dilation of both by isoproterenol support the idea that the VNO is similar to erectile tissue organs and that it may act as a physiological pump.

The vomeronasal organ of the cat.

The vomeronasal organ of the cat was studied macroscopically, by light microscopy and by immunohistochemical techniques. 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.) Examination of series of transverse sections showed that the wall of the vomeronasal duct bears 44 different types of epithelium: simple columnar in the caudal part of the duct, respiratory and receptor respectively on the lateral and medial walls of the middle part of the duct, and stratified squamous rostrally. The pattern of distribution of other soft tissue components was closely associated with that of epithelium types. In areas where the duct wall was lined with receptor epithelium, nerves and connective tissue were present between the epithelium and the medial sheet of the vomeronasal cartilage. Most glands and blood vessels were located lateral to those areas of the duct wall lined with respiratory epithelium. Numerous basal cells were present in the sensory epithelium. Understanding of the distribution of the soft tissue components of this organ may shed light on its function.

Comparative anatomy of the vomeronasal cartilage in mammals: mink, cat, dog, pig, cow and horse.

The vomeronasal cartilages of mink, cat, dog, pig, cow and horse were studied by dissection, microdissection and by means of series of transverse sections. In all the species studied the cartilage is of hyaline type and the medial sheet is well-defined and perfectly moulded to the adjacent bone. However, interspecies differences are apparent in the manner in which the medial sheet associates and eventually fuses with the cartilage of the incisive duct; the morphology of the horse vomeronasal cartilage is particularly distinctive in this respect. The lateral sheet of the vomeronasal cartilage, although always present, has a different arrangement in each species studied. Similarly, the gaps in the lateral sheet (corresponding to the opening of the vomeronasal organ) differ among the species studied in form, location and number.

Structural, morphometric, and immunohistological study of the accessory olfactory bulb in the dog.

BACKGROUND: The study of the morphological, morphometric, and immunohistological characteristics of the accessory olfactory bulb (AOB) in the dog is the main goal of this work. METHODS: Horizontal sections of the AOB where stained by four different methods (haematoxilin/eosin, Tolivia, Nissl, and Bielchowsky). The avidin-biotin-peroxidase complex (ABC) was used, whereas the monoclonal antibodies to neuron-specific enolase, neurofilaments, glial fibrillary acidic protein, and synaptophysin were selected for the immunohistological study. A computer-assisted image analysis was employed in order to define the morphometric characteristics of de AOB. RESULTS: The general morphology of the AOB indicates that it comprises a thick glomerular layer and a thinner internal layer containing mitral/tufted, granular, and glial cells. The mitral/tufted cells have large pale-staining nuclei with intensely staining nucleoli. There does not appear to be a clearly defined granular layer. No reactivity with antibodies to neuron-specific enolase or to neurofilaments was observed in any part of the AOB, but there was some reactivity with an antibody to glial fibrillary acidic protein and widespread reactivity with an antibody to synaptophysin. CONCLUSIONS: The stratification of the AOB is simpler and less well defined than that of the main olfactory bulb (MOB), unlike in rodents in which the structure of the AOB corresponds closely to that of the MOB. According to the scale of Frahm and Bhatnagar (1980. J. Anat., 130: 349-365) the AOB of the adult dog has an intermediate position.