Natural ligands of hamster aphrodisin

The chemical nature of vertebrate pheromones remains largely to be deciphered. Hamster aphrodisin is a rare instance of mammal proteinaceous sexual pheromone. This protein, found in vaginal secretions, facilitates the mounting behaviour of males via activation of a specialized sensory structure named the vomeronasal organ, which activates the accessory olfactory bulb. Since it might carry small pheromonal ligands due to its lipocalin structure, we analysed organic extracts from natural aphrodisin. We identified five predominant compounds specifically bound onto natural aphrodisin as 1-hexadecanol (44.7%), 1-octadecanol (19.5%), Z-9-octadecen-1-ol (18.2%), E-9-octadecen-1-ol (15.4%) and hexadecanoic acid (2.2%). Interestingly, these compounds are also described as part of insect pheromone blends, disclosing the continuing story of amazing coincidences of chemical communication shared by mammals and insects.     

Evidence of an odorant-binding protein in the human olfactory mucus: location,structural characterization,and odorant-binding properties

Odorant-binding proteins (OBPs) are small abundant extracellular proteins belonging to the lipocalin superfamily. They are thought to participate in perireceptor events of odor detection by carrying, deactivating, and/or selecting odorant molecules. Putative human OBP genes (hOBP) have recently been described [Lacazette et al. (2000) Hum. Mol. Genet. 9, 289-301], but the presence of the corresponding proteins remained to be established in the human olfactory mucus. This paper reports the first evidence of such expression in the mucus covering the olfactory cleft, where the sensory olfactory epithelium is located. On the contrary, hOBPs were not observed in the nasal mucus covering the septum and the lower turbinate. To demonstrate the odorant binding activity of these proteins, a corresponding recombinant protein variant, hOBP(IIa)(alpha), was secreted by the yeast Pichia pastoris and thoroughly characterized. It appears as a monomer with one disulfide bond located between C59 and C151, a conservative feature of all other vertebrate OBPs. By measuring the displacement of several fluorescent probes, we show that hOBP(IIa)(alpha) is able to bind numerous odorants of diverse chemical structures, with a higher affinity for aldehydes and large fatty acids. A computed 3D model of hOBP(IIa)(alpha) is proposed and reveals that two lysyl residues of the binding pocket may account for the increased affinity for aldehydes. The relatively limited specificity of hOBP(IIa)(alpha) suggests that other human OBPs are expected to take into account the large diversity of odorant molecules.     

Inflammatory obstruction of the olfactory clefts and olfactory loss in humans: a new syndrome?

The first step in the olfactory perception is the activation by odorants of sensory neurones in the olfactory epithelium. In humans, this sensory epithelium is located at 2 narrow passages, the olfactory clefts, at the upper part of the nasal cavities. Little is known about the physiology of these clefts. We examined, in 34 patients, the impact of obstructed clefts upon detection and postlearning identification of 5 odorants. The location and extension of the obstructions were assessed using endoscopy, CT scans, and MRI. The inflammatory obstruction was usually bilateral, extending anteroposteriorly, and confined to the clefts, with no sign of obstruction or any inflammatory disease in the rest of the nasal cavities and sinuses. When tested with 5 odorants, these patients showed greatly impaired olfaction compared with a group of 73 normosmic subjects. The majority of these 34 patients had sensory deficits equivalent to that found in another group of 41 congenital anosmic patients, where inspection with MRI indicated the lack of olfactory bulbs. This study demonstrates that the olfactory clefts, in human, function as an entity that is different from other regions of the nasal cavity and is the target for local inflammatory events that are apparently not responding to corticoid and antibiotic treatments.     

Scanning electron microscopy and gramicidin patch clamp recordings of microvillous receptor neurons dissociated from the rat vomeronasal organ

Vomeronasal organs from female rats were dissociated and isolated microvillous receptor neurons were studied. The isolated receptor neurons kept the typical bipolar shape which they have in situ as observed by scanning electron microscopy. We applied the perforated patch-clamp technique using the cation-selective ionophore gramicidin on freshly isolated and well differentiated receptor neurons. The mean resting potential was -58+/-14 mV (n=39). The contribution of the sodium pump current to the resting potential was demonstrated by lowering the K+ concentration in the bath or by application of 100 microM dihydro-ouabain. The input resistance was in the range of 1-6 GOmega and depolarizing current pulses of a few pA were sufficient to trigger overshooting action potentials. In voltage clamp conditions a fast transient sodium inward current and a sustained outward potassium current were activated by membrane depolarization. These observations indicate that freshly isolated vomeronasal receptor neurons of rats can be recorded, using gramicidin, with little modification of the intracellular content. Their electrophysiological properties are very similar to those observed in situ. Four out of eight female vomeronasal receptor cells were depolarized by diluted rat male urine.      

Functional Architecture of the Vomeronasal Organ of the Frog (Genus Rana)

Abstract The vomeronasal organ in the frog, genus Rana, is composed of three interconnected cavities; superior, middle and inferior, which are separated from and anterior to the principal olfactory cavity. The superior cavity is found just underneath the external naris and forms a vestibule both for the principal olfactory organ and the vomeronasal organ. The vomeronasal sensory epithelium is located in the medial region of the inferior cavity and contains ciliated cells and microvillous receptor cells. Inspection of microscopic sections of frogs that had been swimming in fluorescent colorants revealed fluorescence on the surface of the vomeronasal organ, but not on that of the olfactory organ. Observations in vivo show that water enters via the external naris by two fissures, one on each side of the movable nasal lid, passes the middle cavity to flow via the sensory epithelium of the inferior cavity. The design of the frog nose makes it possible for this amphibious animal to sample the chemical composition of its environment; above water the frog can inhale air and expose its olfactory organ to volatile substances; in water the vomeronasal organ samples water-borne substances. These new findings are discussed in relation to the air/water interface and the position of the amphibians in the evolution of terrestrial vertebrates.     

Aphrodisin, an aphrodisiac lipocalin secreted in hamster vaginal secretions

Vertebrates communicate through pheromones, which favor biological regulations within each species. Aphrodisin, a protein belonging to the lipocalin superfamily, found in hamster vaginal secretions, is detected by the male accessory olfactory system and induces or facilitates its copulatory behavior. Although much is known about aphrodisin structure, the question of whether aphrodisin bears itself the pheromonal function or is simply a carrier for hydrophobic small pheromones has not been definitely solved. Arguments based on use of recombinant aphrodisin deprived of any natural ligand and its capability to convey hamster pheromonal compounds will be discussed, together with progresses concerning putative natural ligand(s).     

Water Selective Imaging and bSSFP Banding Artifact Correction in Humans and Small Animals at 3T and 7T,Respectively

Introduction

The purpose of this paper is to develop an easy method to generate both fat signal and banding artifact free 3D balanced Steady State Free Precession (bSSFP) images at high magnetic field.

Methods

In order to suppress fat signal and bSSFP banding artifacts, two or four images were acquired with the excitation frequency of the water-selective binomial radiofrequency pulse set On Resonance or shifted by a maximum of 3/4TR. Mice and human volunteers were imaged at 7T and 3T, respectively to perform whole-body and musculoskeletal imaging. “Sum-Of-Square” reconstruction was performed and combined or not with parallel imaging.

Results

The frequency selectivity of 1-2-3-2-1 or 1-3-3-1 binomial pulses was preserved after (3/4TR) frequency shifting. Consequently, whole body small animal 3D imaging was performed at 7T and enabled visualization of small structures within adipose tissue like lymph nodes. In parallel, this method allowed 3D musculoskeletal imaging in humans with high spatial resolution at 3T. The combination with parallel imaging allowed the acquisition of knee images with ~500μm resolution images in less than 2min. In addition, ankles, full head coverage and legs of volunteers were imaged, demonstrating the possible application of the method also for large FOV.

Conclusion

In conclusion, this robust method can be applied in small animals and humans at high magnetic fields. The high SNR and tissue contrast obtained in short acquisition times allows to prescribe bSSFP sequence for several preclinical and clinical applications.     

Odor similarities in structurally related odorants

Similarity assessments against one or several references wereused to estimate "earthiness" in 7 pure cyclic alcohols. The "earthy" odor quality of 3 isomers (I, II, IV) of the moldmetabolite "geosmin" (1, 10dimethyl-9-decalol C₁₂H₂₂O) was foundto be present in 2 other alcohols possessing a partial geosminstructure: exo-2-ethyl-fenchol (C₁₁H₂₂O) (V) and cis-cis 2,6-di-methyl-cyclohexanol(C₈H₁₆O) (VI). All have in common an axial hydroxy group attachedto a 5 or 6 carbon ring with alpha methyls on both sides. The trans-cis and trans-trans isomers of 2,6 dimethyl cyclohexanol(VII, VIII) both with equatorial hydroxy groups, were rankedas different from the above group. Instead their odor resemblesmore that of their aromatic precursor — 2,4 dimethyl phenol(IX) — than that of their axial isomer (VI). The two trans-ring geosmin isomers (I,II) and 2-ethylfenchol(V) were still judged earthy at up to 1000x the lowest concentrationtested. However, the cis-ring geosmin isomer (IV) and the axial cyclohexanol(VI) were perceived by most subjects to change in quality: earthyat lower concentrations but increasingly different at higherones. Discrimination between earthy odorants decreased with decreasingconcentration. Geosmins I, IV and 2 ethyl-fenchol (V) were particularlyconfused. However, discrimination did not appear to be affectedby prior adaptation to geosmin IV. Subjects appeared to be able to focus on the earthy odor qualitycomponent in single odorants IV, V, VI in which several otherquality components could be discerned as well.     

Functional role of receptor neurons in encoding olfactory information

In the present review we have considered the properties of the olfactory receptor neurons and discuss the strategy these cells use to perform their signaling task. Special emphasis is laid on the mechanisms for setting the membrane potential at rest and the mechanisms that the cell can use to respond with action potentials to significant stimuli only. We demonstrate that the firing properties of the receptor neurons depend upon the initial level of the membrane potential. We present the idea that the olfactory glomerulus can function as a unit in olfactory processing. In this perspective the olfactory receptor neuron is a subunit of the olfactory glomerulus. © 1996 John Wiley & Sons, Inc.     

Human genetic polymorphisms in T1R1 and T1R3 taste receptor subunits affect their function

Umami is the typical taste induced by monosodium glutamate (MSG), which is thought to be detected by the heterodimeric G protein-coupled receptor, T1R1 and T1R3. Previously, we showed that MSG detection thresholds differ substantially between individuals and we further showed that nontaster and hypotaster subjects are associated with nonsynonymous single polymorphisms occurring in the T1R1 and T1R3 genes. Here, we show using functional expression that both amino acid substitutions (A110V and R507Q) in the N-terminal ligand-binding domain of T1R1 and the 2 other ones (F749S and R757C), located in the transmembrane domain of T1R3, severely impair in vitro T1R1/T1R3 response to MSG. A molecular model of the ligand-binding region of T1R1/T1R3 provides a mechanistic explanation supporting functional expression data. The data presented here support causal relations between the genotype and previous in vivo psychophysical studies in human evaluating sensitivity to MSG.