Ignacio Salazar

Differential development of binding sites for four lectins in the vomeronasal system of juvenile mouse: from the sensory transduction site to the first relay stage.

Four lectins -the galactose-specific BSI-B(4) (from Bandeiraea simplicifolia), the N-acetyl-galactosamine-specific DBA (from Dolichos biflorus), the L-fucose-specific UEA-I (from Ulex europaeus) and the (oligomeric N-acetylglucosamine)-specific LEA (from Lycopersicum esculentum)- were used to study the vomeronasal organ, vomeronasal nerves and accessory olfactory bulb of the mouse on embryonic days 11, 13, 15, 17 and 19, during the first 3 weeks after birth, at age 25 days, and after reaching maturity. No lectins labelled any structure before the 17th day of gestation, and even on the 19th day staining was sporadic and/or diffuse. During the early postnatal period, the lectin binding patterns differed from those of adults, but the division of the accessory olfactory bulb into anterior, rostral posterior and caudal posterior regions was already present and was shown up by the four lectins in a way that was coherent with the known zone-to-zone correspondence between the apical and basal zones of the sensory epithelium and the anterior and posterior accessory olfactory bulb, respectively. By age 25 days, the staining patterns were essentially those of the adult mouse. BSI-B(4) appears to be specific for the accessory vs. the main olfactory bulb throughout life.

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

Abstract 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.

Anatomy, histochemistry, and immunohistochemistry of the olfactory subsystems in mice.

The four regions of the murine nasal cavity featuring olfactory neurons were studied anatomically and by labeling with lectins and relevant antibodies with a view to establishing criteria for the identification of olfactory subsystems that are readily applicable to other mammals. In the main olfactory epithelium and the septal organ the olfactory sensory neurons (OSNs) are embedded in quasi-stratified columnar epithelium; vomeronasal OSNs are embedded in epithelium lining the medial interior wall of the vomeronasal duct and do not make contact with the mucosa of the main nasal cavity; and in Grünebergs ganglion a small isolated population of OSNs lies adjacent to, but not within, the epithelium. With the exception of Grünebergs ganglion, all the tissues expressing olfactory marker protein (OMP) (the above four nasal territories, the vomeronasal and main olfactory nerves, and the main and accessory olfactory bulbs) are also labeled by Lycopersicum esculentum agglutinin, while Ulex europaeus agglutinin I labels all and only tissues expressing Gαi2 (the apical sensory neurons of the vomeronasal organ, their axons, and their glomerular destinations in the anterior accessory olfactory bulb). These staining patterns of UEA-I and LEA may facilitate the characterization of olfactory anatomy in other species. A 710-section atlas of the anatomy of the murine nasal cavity has been made available on line.

Anatomical, immnunohistochemical and physiological characteristics of the vomeronasal vessels in cows and their possible role in vomeronasal reception

The general morphology of the vomeronasal vessels in adult cows was studied following a classic protocol, including optical, confocal and ultrastructural approaches. This anatomical work was completed immunohistochemically. The vomeronasal organ in cows is well developed, and its vessels are considerable in size. This fact allowed some functional properties of the vomeronasal arteries to be evaluated and, for the first time, their isometric tension to be recorded.

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. Microsc. Res. Tech. 41:492–505, 1998.

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.

The risk of extrapolation in neuroanatomy: the case of the Mammalian vomeronasal system.

The sense of smell plays a crucial role in mammalian social and sexual behaviour, identification of food, and detection of predators. Nevertheless, mammals vary in their olfactory ability. One reason for this concerns the degree of development of their pars basalis rhinencephali, an anatomical feature that has been considered in classifying this group of animals as macrosmatic, microsmatic or anosmatic. In mammals, different structures are involved in detecting odours: the main olfactory system, the vomeronasal system (VNS), and two subsystems, namely the ganglion of Grüneberg and the septal organ. Here, we review and summarise some aspects of the comparative anatomy of the VNS and its putative relationship to other olfactory structures. Even in the macrosmatic group, morphological diversity is an important characteristic of the VNS, specifically of the vomeronasal organ and the accessory olfactory bulb. We conclude that it is a big mistake to extrapolate anatomical data of the VNS from species to species, even in the case of relatively close evolutionary proximity between them. We propose to study other mammalian VNS than those of rodents in depth as a way to clarify its exact role in olfaction. Our experience in this field leads us to hypothesise that the VNS, considered for all mammalian species, could be a system undergoing involution or regression, and could serve as one more integrated olfactory subsystem.

A detailed morphological study of the vomeronasal organ and the accessory olfactory bulb of cats

The organization of the vomeronasal system (VNS) of fetal, newborn, and adult cats was investigated by microdissection and microscopic examination of sections stained conventionally or with lectins (UEA‐1, LEA) or antibodies against proteins Gαi2 (associated with vomeronasal receptor type1) and Gαo (associated with receptor vomeronasal receptor type2). The feline VNS is morphologically similar to that of other mammals. Staining with lectins and anti‐Gαi2 was uniform throughout the sensory epithelium of the vomeronasal organ, and throughout the nervous and glomerular layers of the accessory olfactory bulb (AOB); anti‐Gαo stained no VNS tissue. This organization places the cat together with several other domestic or farm animals (dog, horse, sheep, goat, pig) in a group of mammals with just a single path of communication between the sensory epithelium of the vomeronasal organ and AOB, in contrast to the two‐path model found in rodents and other mammals (in which apical and basal sensory epithelium layers project to rostral and caudal AOB areas, respectively). However, the cat differs from the sheep and pig in that the development of its VNS is still incomplete at birth. Microsc. Res. Tech. 2011.

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

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.