Joerg Strotmann

Rapid activation of alternative second messenger pathways in olfactory cilia from rats by different odorants.

The molecular mechanisms mediating the chemo‐electrical signal transduction in olfactory receptor cells are still elusive. In this study odor induced formation of second messengers in rat olfactory cilia was monitored in a subsecond time range using a rapid kinetic device. Application of micromolar concentration of citralva induced a rapid, transient elevation of the cyclic adenosine monophosphate level, whereas the concentration of inositol trisphosphate was not affected. In contrast, pyrazine caused a rise in the concentration of inositol trisphosphate, not affecting the level of cyclic adenosine monophosphate. Analysis of the kinetic parameter for the odorant induced reaction indicated that apparently two systems are operating simultaneously. The activating effects of odorants appear to be mediated via different G‐proteins. Thus, at least two different second messenger pathways appear to be involved in olfactory signal transduction.

Two classes of olfactory receptors in xenopus laevis

Xenopus laevis possess a gene repertoire encoding two distinct classes of olfactory receptors: one class related to receptors of fish and one class similar to receptors of mammals. Sequence comparison indicates that the fish-like receptors represent closely related members of only two subfamilies, whereas mammalian-like receptors are more distantly related, most of them representing a different subfamily. The fish-like receptor genes are exclusively expressed in the lateral diverticulum of the frogs nose, specialized for detecting water-soluble odorants, whereas mammalian-like receptors are expressed in sensory neurons of the main diverticulum, responsible for the reception of volatile odors.

Rostro-caudal patterning of receptor-expressing olfactory neurones in the rat nasal cavity

The rostro-caudal extent of odorant receptor expression zones in the rat olfactory epithelium was analysed by means of in situ hybridization. Three broad non-overlapping zones were identified that extended along almost the entire anterior-posterior axis; each zone was composed of several separate bands running anterior to posterior throughout the olfactory epithelium. Superimposed onto these broad zones was the expression area of a particular receptor subtype (OR37); it was restricted to a small region of the epithelial sheet with a high density of reactive neurones in the centre and declining numbers towards the periphery of the region. A quantitative evaluation of the reactive cells revealed that, despite their diferent distribution patterns, all receptor subtypes were expressed in an equal number of neurones.

Olfactory neurones expressing distinct odorant receptor subtypes are spatially segregated in the nasal neuroepithelium.

In situ hybridization techniques have been employed to explore the olfactory epithelium of the rat for the distribution of odorant receptor gene transcripts. We demonstrate that olfactory neurone subpopulations expressing distinct receptor subtypes are spatially segregated within the olfactory epithelium. A compartmentalization of the neuroepithelium into distinct expression zone is apparent, cells expressing a specific receptor are randomly distributed within a given zone. Structurally related receptor subtypes share a common distribution pattern.

Olfactory Receptor Proteins in Axonal Processes of Chemosensory Neurons

Olfactory receptors are supposed to act not only as molecular sensors for odorants but also as cell recognition molecules guiding the axons of olfactory neurons to their appropriate glomerulus in the olfactory bulb. This concept implies that olfactory receptor proteins are located in sensory cilia and in the axons. To approach this critical issue, antibodies were generated against two peptides, one derived from olfactory receptor mOR256-17, one derived from the “mOR37” subfamily. By means of immunohistochemistry and double-labeling studies using transgenic mouse lines as well as Western blot analyses, it was demonstrated that the newly generated antibodies specifically recognized the receptor proteins. To scrutinize the hypothesis that olfactory receptor proteins may also be present in the axonal processes and the nerve terminals, serial sections through the olfactory bulb were probed with the antibodies. Two glomeruli in each bulb were stained by anti-mOR256-17, one positioned in the medial, one in the lateral hemisphere. Fiber bundles approaching the glomeruli through the outer nerve layer also displayed intense immunofluorescence. A similar picture emerged for the antibody anti-mOR37, a small number of glomeruli in the ventral domain of the bulb was stained. On serial sections through the olfactory bulb of mOR37-transgenic mouse lines, double-labeling experiments demonstrated that distinct immunoreactive glomeruli corresponded to glomeruli that were targeted by neurons expressing a particular member of the mOR37 receptor subfamily. These data indicate that olfactory receptor (OR) proteins are indeed present in the axonal processes and nerve terminals of olfactory sensory neurons, thus supporting the notion that ORs may participate in the molecular processes underlying the fasciculation and targeting of olfactory axons.

Cloning of biogenic amine receptors from moths (Bombyx mori and Heliothis virescens)

Based on the similarity of genes which code for guanine-nucleotide binding protein- (G-protein-) coupled receptors, cDNA clones encoding new members of the receptor family have been isolated from Bombyx mori and Heliothis virescens. The deduced protein structures exhibit highest similarity to tyramine/octopamine and serotonin receptors of Drosophila. One of the receptor clones (K50Hel) was permanently expressed in the mammalian cell line LLC-PK1. In stimulation experiments its responded to octopamine leading to an inhibition of adenylate cyclase activity in a dose-dependent manner. Pharmacological studies revealed a higher affinity for mianserin than for yohimbine suggesting, that the K50Hel clone encoded a neuronal type 3 octopamine receptor. As revealed by in situ hybridization, this receptor type is expressed in the central nervous system and antennae of moth.

The sense of smell: multiple olfactory subsystems

Abstract.The mammalian olfactory system is not uniformly organized but consists of several subsystems each of which probably serves distinct functions. Not only are the two major nasal chemosensory systems, the vomeronasal organ and the main olfactory epithelium, structurally and functionally separate entities, but the latter is further subcompartimentalized into overlapping expression zones and projection-related subzones. Moreover, the populations of ‘OR37’ neurons not only express a unique type of olfactory receptors but also are segregated in a cluster-like manner and generally project to only one receptor-specific glomerulus. The septal organ is an island of sensory epithelium on the nasal septum positioned at the nasoplatine duct; it is considered as a ‘mini-nose’ with dual function. A specific chemosensory function of the most recently discovered subsystem, the so-called Grueneberg ganglion, is based on the expression of olfactory marker protein and the axonal projections to defined glomeruli within the olfactory bulb. This complexity of distinct olfactory subsystems may be one of the features determining the enormous chemosensory capacity of the sense of smell.

Signaling in the Chemosensory Systems

Abstract.The mammalian olfactory system is not uniformly organized but consists of several subsystems each of which probably serves distinct functions. Not only are the two major nasal chemosensory systems, the vomeronasal organ and the main olfactory epithelium, structurally and functionally separate entities, but the latter is further subcompartimentalized into overlapping expression zones and projection-related subzones. Moreover, the populations of ‘OR37’ neurons not only express a unique type of olfactory receptors but also are segregated in a cluster-like manner and generally project to only one receptor-specific glomerulus. The septal organ is an island of sensory epithelium on the nasal septum positioned at the nasoplatine duct; it is considered as a ‘mini-nose’ with dual function. A specific chemosensory function of the most recently discovered subsystem, the so-called Grueneberg ganglion, is based on the expression of olfactory marker protein and the axonal projections to defined glomeruli within the olfactory bulb. This complexity of distinct olfactory subsystems may be one of the features determining the enormous chemosensory capacity of the sense of smell.

Mammalian olfactory receptors

Perception of chemical stimuli from the environment is essential to most animals; accordingly, they are equipped with a complex olfactory system capable of receiving a nearly unlimited number of odorous substances and pheromones. This enormous task is accomplished by olfactory sensory neurons (OSNs) arranged in several chemosensory compartments in the nose. The sensitive and selective responsiveness of OSNs to odorous molecules and pheromones is based on distinct receptors in their chemosensory membrane; consequently, olfactory receptors play a key role for a reliable recognition and an accurate processing of chemosensory information. They are therefore considered as key elements for an understanding of the principles and mechanisms underlying the sense of smell. The repertoire of olfactory receptors in mammals encompasses hundreds of different receptor types which are highly diverse and expressed in distinct subcompartments of the nose. Accordingly, they are categorized into several receptor families, including odorant receptors (ORs), vomeronasal receptors (V1Rs and V2Rs), trace amine-associated receptors (TAARs), formyl peptide receptors (FPRs), and the membrane guanylyl cyclase GC-D. This large and complex receptor repertoire is the basis for the enormous chemosensory capacity of the olfactory system.

Cells in the vomeronasal organ express odorant receptors but project to the accessory olfactory bulb

Recent evidence indicates that the vomeronasal organ (VNO) of mice not only responds to pheromones but also to odorants. To analyze whether genes encoding odorant receptors (ORs) are expressed in the VNO, reverse transcriptase‐polymerase chain reaction analyses were performed. These led to the identification of 44 different OR genes, comprising class‐I and class‐II receptors. The genes encoding these receptors were scattered over several gene clusters. The respective OR genes were concomitantly expressed in cells of the main olfactory epithelium (MOE). Although the cells in the MOE were zonally distributed, no such patterns were displayed in the VNO. Cells expressing ORs in the VNO were positive for the TRP2‐channel and Gαi, a marker for vomeronasal neurons of the apical layer. In transgenic mice, which coexpress histological markers with the receptor mOR18‐2, characteristic morphological differences between cells expressing this receptor in the VNO compared with the MOE became evident. Visualizing the axonal processes of VNO cells expressing distinct ORs revealed that they project to the accessory olfactory bulb (AOB). Axon fibers were visible exclusively in the anterior subdomain; here, they converged into glomerular‐like structures positioned at the very rostral tip of the AOB. The findings that a set of ORs is expressed in cells located in the apical layer of the VNO with typical features of VNO sensory neurons that project their axons to the anterior part of the AOB suggest that this population of sensory cells may be considered as a unique facet of the complex chemosensory system. J. Comp. Neurol. 498:476–490, 2006.