Subfamily of Olfactory Receptors Characterized by Unique Structural Features and Expression Patterns

Abstract: The complex chemospecificity of the olfactory system is probably due to the large family of short-looped, heptahelical receptor proteins expressed in neurons widely distributed throughout one of the several zones within the nasal neuroepithelium. In this study, a subfamily of olfactory receptors has been identified that is characterized by distinct structural features as well as a unique expression pattern. Members of this receptor family are found in mammals, such as rodents and opossum, but not in lower vertebrates. All identified subtypes comprise an extended third extracellular loop that exhibits amphiphilic properties and contains numerous charged amino acids in conserved positions. Olfactory sensory neurons expressing these receptor types are segregated in small clusters on the tip of central turbinates, thus representing a novel pattern of expression for olfactory receptors. In mouse, genes encoding the new subfamily of receptors were found to be harbored within a small contiguous segment of genomic DNA. Based on species specificity as well as the unique structural properties and expression pattern, it is conceivable that the novel receptor subfamily may serve a special function in the olfactory system of mammals.     

What is it like to be a rat? Rat sensory perception and its implications for experimental design and rat welfare

This review of rat sensory perception spans eight decades of work conducted across diverse research fields. It covers rat vision, audition, olfaction, gustation, and somatosensation, and describes how rat perception differs from and coincides with ours. As Nagel's seminal work (1974) implies, we cannot truly know what it is like to be a rat, but we can identify and acknowledge their perceptual biases. These primarily nocturnal rodents are extremely sensitive to light, with artificial lighting frequently causing retinal degeneration, and their vision extends into the ultraviolet. Their olfactory sensitivity and ultrasonic hearing means they are influenced by environmental factors and conspecific signals that we cannot perceive. Rat and human gustation are similar, being opportunistic omnivores, yet this sense becomes largely redundant in the laboratory, where rodents typically consume a single homogenous diet. Rat somatosensation differs from ours in their thigmotactic tendencies and highly sensitive, specialised vibrissae. Knowledge of species-specific perceptual abilities can enhance experimental designs, target resources, and improve animal welfare. Furthermore, the sensory environment has influences from neurone to behaviour, so it can not only affect the senses directly, but also behaviour, health, physiology, and neurophysiology. Research shows that environmental enrichment is necessary for normal visual, auditory, and somatosensory development. Laboratory rats are not quite the simple, convenient models they are sometimes taken for; although very adaptable, they are complex mammals existing in an environment they are not evolutionarily adapted for. Here, many important implications of rat perception are highlighted, and suggestions are made for refining experiments and housing.     

Dopaminergic systems in the European eel: characterization,brain distribution,and potential role in migration and reproduction

In fish like in mammals, dopamine (DA) is a major catecholaminergic neurotransmitter that contributes to many functions of the nervous system like sensory perception, tuning of sensori-motor cues, and hypothalamic and pituitary functions. In the eel, DA inhibits gonadal development, and juvenile silver eels remain blocked at a prepubertal stage if their reproductive migration does not occur. From data in other teleosts and vertebrates, it is suggested that DA would be involved also in the last steps of eel reproduction (oocyte maturation, ovulation, and spermiation) as well as in eel reproductive migration (locomotion and olfaction). Investigating dopaminergic systems in the eel may help in understanding the mechanisms of its complex life cycle and provide new data for its conservation and reproduction. In this article we review the biosynthesis and catabolism of catecholamines and discuss available methods to investigate brain dopaminergic systems in vertebrates and their application to the eel. Immunocytochemistry, in situ hybridization, and different tracing methods are used to map dopaminergic neurons and projections in the brain and pituitary and infer their potential functions. Moreover, variations in dopaminergic activity may be approached by means of quantitative methods like quantitative real-time RT-PCR and HPLC. These tools are currently used to study dopaminergic systems in the eel brain, their anatomy, regulation, and potential roles with special emphasis on the regulation of reproduction and reproductive migration. Guest editors: S. Dufour, E. Prévost, E. Rochard & P. Williot Fish and diadromy in Europe (ecology, management, conservation)     

Crystal structure of Apis mellifera OBP14, a C-minus odorant-binding protein, and its complexes with odorant molecules

Apis mellifera (Amel) relies on its olfactory system to detect and identify new-sources of floral food. The Odorant-Binding Proteins (OBPs) are the first proteins involved in odorant recognition and interaction, before activation of the olfactory receptors. The Amel genome possess a set of 21 OBPs, much fewer compared to the 60-70 OBPs found in Diptera genomes. We have undertaken a structural proteomics study of Amel OBPs, alone or in complex with odorant or model compounds. We report here the first 3D structure of a member of the C-minus class OBPs, AmelOBP14, characterized by only two disulfide bridges of the three typical of classical OBPs. We show that AmelOBP14 possesses a core of 6 α-helices comparable to that of classical OBPs, and an extra exposed C-terminal helix. Its binding site is located within this core and is completely closed. Fluorescent experiments using 1-NPN displacement demonstrate that AmelOBP14 is able to bind several compounds with sub micromolar dissociation constants, among which citralva and eugenol exhibit the highest affinities. We have determined the structures of AmelOBP14 in complex with 1-NPN, eugenol and citralva, explaining their strong binding. Finally, by introducing a double cysteine mutant at positions 44 and 97, we show that a third disulfide bridge was formed in the same position as in classical OBPs without disturbing the fold of AmelOBP14.     

Anatomical organization of antennal-lobe glomeruli in males and females of the scarab beetle Holotrichia diomphalia (Coleoptera: Melolonthidae)

The glomerular organization of the primary olfactory brain center, the antennal lobe, was studied in males and females of Holotrichia diomphalia adults using serial histological sections labeled by the reduced silver-stain technique. The results revealed an apparent sexual dimorphism. Whereas an enlarged cap-shaped glomerulus was found at the antennal nerve entrance into the antennal lobe in males, no such unit was present in females. Also the size of the antennal lobe differed between the sexes, the antennal lobe of males being larger than that of females. We estimated the total number of glomeruli at approximately 60 units in the female antennal lobe. In males, we could discriminate only those glomeruli that were located in the anterior area of the antennal lobe.     

New insights into the mechanism of odorant detection by the malaria-transmitting mosquito Anopheles gambiae

Anopheles gambiae mosquitoes that transmit Plasmodium falciparum malaria use a series of olfactory cues present in human sweat to locate their hosts for a blood meal. Recognition of these odor cues occurs through the interplay of odorant receptors and odorant-binding proteins (OBPs) that bind to odorant molecules and transport and present them to the receptors. Recent studies have implicated potential heterodimeric interactions between two OBPs, OBP1 and OBP4, as important for perception of indole by the mosquito (Biessmann, H., Andronopoulou, E., Biessmann, M. R., Douris, V., Dimitratos, S. D., Eliopoulos, E., Guerin, P. M., Iatrou, K., Justice, R. W., Kröber, T., Marinotti, O., Tsitoura, P., Woods, D. F., and Walter, M. F. (2010) PLoS ONE 5, e9471; Qiao, H., He, X., Schymura, D., Ban, L., Field, L., Dani, F. R., Michelucci, E., Caputo, B., della Torre, A., Iatrou, K., Zhou, J. J., Krieger, J., and Pelosi, P. (2011) Cell. Mol. Life Sci. 68, 1799–1813). Here we present the 2.0 Å crystal structure of the OBP4-indole complex, which adopts a classical odorant-binding protein fold, with indole bound at one end of a central hydrophobic cavity. Solution-based NMR studies reveal that OBP4 exists in a molten globule state and binding of indole induces a dramatic conformational shift to a well ordered structure, and this leads to the formation of the binding site for OBP1. Analysis of the OBP4-OBP1 interaction reveals a network of contacts between residues in the OBP1 binding site and the core of the protein and suggests how the interaction of the two proteins can alter the binding affinity for ligands. These studies provide evidence that conformational ordering plays a key role in regulating heteromeric interactions between OBPs.     

Endogenous oxytocin is necessary for preferential Fos expression to male odors in the bed nucleus of the stria terminalis in female Syrian hamsters

Successful reproduction in mammals depends on proceptive or solicitational behaviors that enhance the probability of encountering potential mates. In female Syrian hamsters, one such behavior is vaginal scent marking. Recent evidence suggests that the neuropeptide oxytocin (OT) may be critical for regulating this behavior. Blockade of OT receptors in the bed nucleus of the stria terminalis (BNST) or the medial preoptic area (MPOA) decreases vaginal marking responses to male odors; lesion data suggest that BNST, rather than MPOA, mediates this effect. However, how OT interacts with sexual odor processing to drive preferential solicitation is not known. To address this issue, intact female Syrian hamsters were exposed to male or female odors and their brains processed for immunohistochemistry for Fos, a marker of recent neuronal activation, and OT. Additional females were injected intracerebroventricularly (ICV) with an oxytocin receptor antagonist (OTA) or vehicle, and then tested for vaginal marking and Fos responses to sexual odors. Colocalization of OT and Fos in the paraventricular nucleus of the hypothalamus was unchanged following exposure to male odors, but decreased following exposure to female odors. Following injections of OTA, Fos expression to male odors was decreased in BNST, but not in MPOA or the medial amygdala (MA). Fos expression in BNST may be functionally relevant for vaginal marking, given that there was a positive correlation between Fos expression and vaginal marking for BNST, but not MPOA or MA. Together, these data suggest that OT facilitation of neuronal activity in BNST underlies the facilitative effects of OT on solicitational responses to male odors.     

Discrimination of chemical stimuli in conspecific fecal pellets by a visually adept iguanid lizard,Crotaphytus collaris

Iguanid lizards are known for visual acuity and a diminished vomeronasal organ, which has led to mixed conclusions on whether iguanids use chemical cues. The collared lizard, Crotaphytus collaris, is a territorial iguanid that lives in open rocky habitats. Fecal pellets placed prominently on open rocky perches may provide an ideal mechanism for intraspecific chemical signaling. In order to determine whether collared lizards can discriminate between chemical stimuli found in conspecific fecal pellets, we collected 24 males and 25 females to analyze sex-specific behavioral responses via tongue-flicks and a newly observed behavior for the species, gular pumps, to cotton swabs containing water, cologne, chemical stimuli from conspecific male and female fecal pellets, and the lizard’s own fecal pellet. Both sexes were able to discriminate chemical stimuli from water via at least one behavior. Male collared lizards exhibited greater rates of response (tongue-flick and gular pumps) toward male fecal pellets when compared to the negative water control. Our results also suggest individuals may be able to discriminate between fecal pellets, as indicated by generally greater (but non-significant) counts of male tongue-flick responses to male fecal pellets when compared to their own. Collared lizard chemical discrimination appears to utilize tongue-flick and gular pump behaviors, possibly associated with distinct chemosensory modes (vomerolfaction and olfaction). Based on this study, we suggest that chemical signals may play a greater role in intraspecific communication than previously thought in this highly visual lizard.     

Activity modulation in cockroach sensillum: the role of octopamine

The plasticity of sensory perception is provided partially by modulation of receptor cells. The electrical activity of American cockroach chemoreceptor cells in response to sex pheromone was measured under the influence of octopamine treatment and tracheal anoxia. Both experimental procedures caused decreased electroantennograms but affected spike activity differently: octopamine treatment increased firing rate, whereas anoxia decreased it. Spike frequency under octopamine treatment was elevated in response to pheromone stimulation and at background activity. Experiments with perfusion of isolated antennae showed a direct effect of octopamine on spike activity of pheromone sensilla, and excluded the possibility of indirect effects via octopamine-dependent release of other biologically active substances. The suggested mechanism of octopamine action is receptor cell membrane depolarization.     

Preventing physical interactions between parturient ewes and their neonate differentially impairs the development of maternal responsiveness and selectivity depending on maternal experience

The present study investigated whether deprivation of nursing or of any physical interaction with the lamb impairs the onset of maternal responsiveness and the establishment of selectivity in primiparous and multiparous ewes. Just after parturition, ewes could interact freely with their lamb, or could only lick their lamb but not nurse them, or could neither lick nor nurse them for the first 4 h. At 4 h, mothers were successively observed for 3 min with an alien lamb and then with their own lamb. Only 5/14 primiparous mothers accepted their own lamb following deprivation of all physical contact whereas all multiparous mothers did so (19/19). Also, deprivation of all physical contact resulted in a significantly higher proportion of multiparous mothers accepting an alien lamb (6/18) than following deprivation of nursing only (0/19) indicating an impairment in selectivity. Finally, there was no indication that preventing only nursing led to a deficit either in maternal responsiveness or selectivity, neither in primiparous nor in multiparous ewes. These findings underline the importance of perception of olfactory cues from amniotic fluids through licking and/or physical contact and interacting with the lamb for the development of maternal responsiveness and selectivity.