Molecule capture by olfactory antennules: Mantis shrimp

 A critical step in the process of olfaction is the movement of odorant molecules from the environment to the surface of a chemosensory structure. Many marine crustaceans capture odorant molecules with arrays of chemosensory sensilla (aesthetascs) on antennules that they flick through the water. We developed a model to calculate molecule flux to the surfaces of aesthetascs in order to study how the size, aesthetasc spacing, and flick kinematics of olfactory antennules affect their performance in capturing molecules from the surrounding water. Since the three-dimensional geometry of an aesthetasc-bearing antennule is complex, dynamically-scaled physical models can often provide an efficient method of determining the fluid velocity field through the array. Here we present a method to optimize the incorporation of such measured velocity vector fields into a numerical simulation of the advection and diffusion of odorants to aesthetasc surfaces. Furthermore, unlike earlier models of odorant interception by antennae, our model incorporates odorant concentration distributions that have been measured in turbulent ambient flows. By applying our model to the example of the olfactory antennules of mantis shrimp, we learned that flicking velocity can have profound effects on odorant flux to the aesthetascs if they operate in the speed range in which the leakiness of the gaps between the aesthetascs to fluid movement is sensitive to velocity. This sensitivity creates an asymmetry in molecule fluxes between outstroke and return stroke, which results in an antennule taking discrete samples in space and time, i.e. “sniffing”. As stomatopods grow and their aesthetasc Reynolds number increases, the aesthetasc arrangement on the antennule changes in a way that maintains these asymmetries in leakiness and molecule flux between the outstroke and return stroke, allowing the individual to continue to take discrete samples as it develops. Received: 24 May 2000 / Revised version: 8 May 2001 / Published online: 7 December 2001     

Identification of chemosensory sensilla mediating antennular flicking behavior in Panulirus argus, the Caribbean spiny lobster

Crustaceans sample odorants by a rapid series of flicks of the two flagella composing the distal segments of each of the paired antennules. The lateral flagella contain aesthetasc sensilla that house unimodal chemosensory neurons. Nine types of nonaesthetasc setae with putative chemosensory and mechanosensory functions are distributed on the lateral and medial flagella. Sensory neurons in aesthetascs and nonaesthetasc sensilla terminate in separate regions of the brain, the olfactory lobe, and the lateral antennular neuropil, resulting in two odorant-processing pathways. Distilled water ablation of flagella and excision of specific setae were used to identify chemosensory sensilla mediating antennular flick behavior in Panulirus argus. The flick rates of sham-ablated and ablated or excised lobsters toward squid extract were compared. Complete attenuation of flick response to squid extract occurred as a result of (1) distilled water ablation of lateral flagella, (2) excision of aesthetascs and asymmetric sensilla, and (3) excision of aesthetascs. Distilled water ablation of medial flagella resulted in a mean flick rate 52% of that observed for sham-ablated lobsters toward squid extract. Flicking was unaffected by excision of asymmetric, guard, or companion sensilla. We propose that odorant mediation of flicking behavior requires both the aesthetasc and nonaesthetasc pathways.     

Fluid dynamic design of lobster olfactory organs: high speed kinematic analysis of antennule flicking by Panulirus argus

Many organisms use olfactory appendages bearing arrays of microscopic hairs to pick up chemical signals from the surrounding water or air. We report a morphometric and high speed kinematic analysis of the olfactory organs (lateral flagella of the antennules, which bear chemosensory aesthetasc hairs) of the spiny lobster, Panulirus argus. Panulirus argus sample specific locations by executing a rapid series of antennule flicks at one position, moving the antennule to a different spot and then performing another series of flicks. Odorant delivery to an aesthetasc depends on the water motion near it, which depends on its Reynolds number (Re, proportional to both the diameter and speed of the hair). High speed video enabled us to resolve that during a series of flicks, an antennule moves down rapidly (aesthetasc Re = 2) and up more slowly (Re = 0.5), pausing briefly ( approximately 0.54 s) before the next downstroke. The antennules of P. argus operate in a range of Re values and inter-aesthetasc spacings in which penetration of fluid between the hairs in an array is especially sensitive to changes in speed. Therefore, when antennules flick 'old' water is flushed out of the aesthetasc array during the leaky downstroke and is not picked up again during the less leaky upstroke, hence the antennules can take discrete samples. Thus, by operating in this critical Re range these antennules should be particularly effective at sniffing.     

Morphological characteristics facilitating stimulus access and removal in the olfactory organ of the spiny lobster, Panulirus argus: insight from the design

The olfactory organ (antennule) of the spiny lobster, Panulirusargus, has from 1000–2000 olfactory sensilla (aesthetascs)which are grouped in a dense tuft along the distal portion ofthe lateral filament. This assemblage of aesthetascs, togetherwith other associated sensilla, forms a substantial boundarylayer through which odor stimuli must diffuse in moving to andfrom the aesthetascs. Periodic flicking of the antennule, abehavior analogous to sniffing in certain vertebrate species,is considered to be a means of reducing the thickness of thisboundary layer. In this report we describe the structure ofthe aesthetasc tuft and examine certain of its dynamic properties.We propose that the unique configuration of the aesthetasces,together with their orientation, serves to channel water flowbetween these sensilla during a flick, thereby reducing diffusiondistances and consequently facilitating the access and removalof odor stimuli in a rapid, synchronized manner. The functionalsignificance of this and other design features of the aesthetasctuft is considered in light of the current understanding offundamental olfactory process.     

The fluid mechanics of arthropod sniffing in turbulent odor plumes

Many arthropods capture odorant molecules from the environment using antennae or antennules bearing arrays of chemosensory hairs. The penetration of odorant-carrying water or air into the spaces between these chemosensory hairs depends on the speed at which they are moved through the surrounding fluid. Therefore, antennule flicking by crustaceans and wing fanning by insects can have a profound impact on the odorant encounter rates of the chemosensory sensilla they bear; flicking and fanning are examples of sniffing. Odors are dispersed in the environment by turbulent wind or water currents. On the scale of an antenna or antennule, an odor plume is not a diffuse cloud but rather is a series of fine filaments of scent swirling in odor-free water. The spatiotemporal pattern of these filaments depends on distance from the odor source. The physical interaction of a hair-bearing arthropod antennule with the surrounding fluid affects the temporal patterns of odor concentration an animal intercepts when it sniffs in a turbulent odor plume.     

Physiological characterization of antennular flicking reflexes in the crayfish

The cellular substrates of antennular flicking behavior in the crayfish Procambarus clarkii were investigated. Flicking involves fast downward movements of the external filament of each biramous antennule (1st antenna), and is mediated by phasic contractions of a short muscle, the external filament depressor. Phasic contractions of the external filament depressor depend upon stereotyped impulse bursts in a single motorneuron (P1). These bursts have a characteristic impulse frequency profile that is consistent upon successive occurrences. The temporal characteristics of the impulse burst suggest that the central depolarizations generating each burst may be similar to driver potentials described for motor neurons in crustacean cardiac ganglia. Responses of the external filament to odorants have a long latency and are characterized by repetitive bursts and tonic activity in some external filament depressor fibers. Tonic activity in a slowly contracting muscle, the antennular depressor muscle, is also evoked by chemical stimulation. Flicking is consistently evoked only by mechanical or hydrodynamic stimulation of the cephalothorax, antennae and antennules. The sensitivity and short latency of the hydrodynamic antennule-generated flick reflex is consistent with the sensitivity of rapidly conducting, hydrodynamically activated mechanoreceptor neurons in both antennular filaments. I propose that antennular flicking, which has been shown to enhance the dynamic response characteristics of olfactory receptor neurons on the external antennular filament, has evolved as a response to the turbulence associated with fluid movement, within which chaotic odorant concentration fronts may be imbedded. Accepted: 23 October 1996     

Identification of chemosensory sensilla activating antennular grooming behavior in the Caribbean spiny lobster,Panulirus argus

Crustaceans such as crabs and lobsters clean or 'groom' their olfactory organ, the antennule, by wiping it through a pair of mouthpart appendages, the third maxillipeds. In the lobster, only a few chemicals found in prey extracts, especially glutamate, elicit grooming. Chemosensory input driving grooming is likely to be mediated via sensilla located on antennules and third maxillipeds. Chemosensory antennular sensilla are innervated by neurons with central projections either to the glomerular olfactory lobe (aesthetasc sensilla) or to non-glomerular antennular neuropils (nonaesthetasc sensilla). By selectively ablating the chemosensory sensilla on the antennules and the third maxillipeds we have determined that the aesthetascs are necessary and sufficient to drive grooming behavior. Chemosensory activation of antennular grooming behavior likely follows a 'labeled-line' model in that aesthetasc neurons tuned to glutamate provide adequate input via the olfactory lobe to motor centers in the brain controlling antennular movements.     

Ultrastructure of the peduncular aesthetascs of Speleonectes tanumekes (Crustacea, Remipedia) in comparison with crustaceans from stressful environments

Abstract. Osmotic stress associated with the freshwater environment and desiccation stress associated with the terrestrial environment may have a shortening effect on the length of the innervation of crustacean aesthetascs. Physical stress of the littoral environment may have a similar effect on the length of the cuticular portion of aesthetascs. The aesthetascs of crustaceans that inhabit these environments share a similar ultrastructural feature, which may help animals cope with these environmental stresses. This ultrastructural feature, the position of the basal bodies proximal to the lumen of the aesthetasc, is absent from the aesthetascs of crustaceans that occur in the typical marine environment. Interestingly, the ultrastructural feature associated with these stressful habitats is present in the peduncular aesthetascs of the remipede Speleonectes tanumekes , even though the environmental stresses that may invoke the reduction of aesthetascs are absent in the marine-cave environment where this animal occurs. The importance of the sensitivity of aesthetascs for survival in this lightless environment may result in a selective pressure that favors basal bodies to be positioned proximal to the lumen of the aesthetasc.     

The modality of the dominance signal in snapping shrimp (Alpheus heterochaelis) and the corresponding setal types on the antennules

Previous behavioural experiments showed that snapping shrimp lacking lateral antennular filaments, i.e. without chemosensory aesthetascs, lose the ability to distinguish between conspecifics that are inexperienced in fighting and former winners. A chemosensory dominance signal was assumed to be present, although other receptors unique to the lateral filaments may have been responsible for the behavioural changes. In the present study, the antennules of snapping shrimp were examined for differences between the lateral and medial antennule filaments to identify the modality of the dominance signal. We found six different types of setae and two types of pores. A new probably bimodal setal type is described, the broad long simple seta. Only the chemosensory aesthetascs and their associated hydrosensory companion setae are unique to the lateral filament. Thus we conclude that the dominance signal is chemical, because a hydrodynamic signal would be also received by the simple setae distributed on both filaments.     

The modality of the dominance signal in snapping shrimp (Alpheus heterochaelis) and the corresponding setal types on the antennules

Previous behavioural experiments showed that snapping shrimp lacking lateral antennular filaments, i.e. without chemosensory aesthetascs, lose the ability to distinguish between conspecifics that are inexperienced in fighting and former winners. A chemosensory dominance signal was assumed to be present, although other receptors unique to the lateral filaments may have been responsible for the behavioural changes. In the present study, the antennules of snapping shrimp were examined for differences between the lateral and medial antennule filaments to identify the modality of the dominance signal. We found six different types of setae and two types of pores. A new probably bimodal setal type is described, the broad long simple seta. Only the chemosensory aesthetascs and their associated hydrosensory companion setae are unique to the lateral filament. Thus we conclude that the dominance signal is chemical, because a hydrodynamic signal would be also received by the simple setae distributed on both filaments.     

Learned odor discrimination in Drosophila without combinatorial odor maps in the antennal lobe

A unifying feature of mammalian and insect olfactory systems is that olfactory sensory neurons (OSNs) expressing the same unique odorant-receptor gene converge onto the same glomeruli in the brain [1-7]. Most odorants activate a combination of receptors and thus distinct patterns of glomeruli, forming a proposed combinatorial spatial code that could support discrimination between a large number of odorants [8-11]. OSNs also exhibit odor-evoked responses with complex temporal dynamics [11], but the contribution of this activity to behavioral odor discrimination has received little attention [12]. Here, we investigated the importance of spatial encoding in the relatively simple Drosophila antennal lobe. We show that Drosophila can learn to discriminate between two odorants with one functional class of Or83b-expressing OSNs. Furthermore, these flies encode one odorant from a mixture and cross-adapt to odorants that activate the relevant OSN class, demonstrating that they discriminate odorants by using the same OSNs. Lastly, flies with a single class of Or83b-expressing OSNs recognize a specific odorant across a range of concentration, indicating that they encode odorant identity. Therefore, flies can distinguish odorants without discrete spatial codes in the antennal lobe, implying an important role for odorant-evoked temporal dynamics in behavioral odorant discrimination.     

A computational study of odorant transport and deposition in the canine nasal cavity: implications for olfaction

Olfaction begins when an animal draws odorant-laden air into its nasal cavity by sniffing, thus transporting odorant molecules from the external environment to olfactory receptor neurons (ORNs) in the sensory region of the nose. In the dog and other macrosmatic mammals, ORNs are relegated to a recess in the rear of the nasal cavity that is comprised of a labyrinth of scroll-like airways. Evidence from recent studies suggests that nasal airflow patterns enhance olfactory sensitivity by efficiently delivering odorant molecules to the olfactory recess. Here, we simulate odorant transport and deposition during steady inspiration in an anatomically correct reconstructed model of the canine nasal cavity. Our simulations show that highly soluble odorants are deposited in the front of the olfactory recess along the dorsal meatus and nasal septum, whereas moderately soluble and insoluble odorants are more uniformly deposited throughout the entire olfactory recess. These results demonstrate that odorant deposition patterns correspond with the anatomical organization of ORNs in the olfactory recess. Specifically, ORNs that are sensitive to a particular class of odorants are located in regions where that class of odorants is deposited. The correlation of odorant deposition patterns with the anatomical organization of ORNs may partially explain macrosmia in the dog and other keen-scented species.     

Fine structure of the aesthetasc hairs of Coenobita compressus Edwards

The aesthetascs, short thin-walled pegs on the antennule flagella of Coenobita clypeatus, a terrestrial hermit crab, are similar to those of other decapod crustacea in containing the dendrites of many bipolar neurons whose cell bodies are grouped in spindle-shaped masses beneath the bases of each hair. The dendrites contain rootlets, basal bodies, and cilia, which divide dichotomously before entering the aesthetasc, so that within the hair, each cilium becomes represented by a group of slender branches. The aesthetascs themselves are short, blunt, and partially recumbent so that each has an exposed and an unexposed side. The cuticle on the exposed side is thinner and more tenuous than that on the protected side, and the dendrite branches are concentrated just underneath. The protected side, on the other hand, is lined with nondendritic supporting cells, and the cuticle is thicker, more lamellar, and probably less permeable. All dendritic elements proximal to the dendrite branches are enclosed within the main body of the antennular flagellum, and the initial segments of the cilia lie within a vacuole. In these respects, the aesthetascs of Coenobita resemble the thin-walled pegs on insect antennae more than they do those of the marine decapods thus far examined. This convergence in the terrestrial forms may be in response to the need to conserve water.     

Morphology of stomatopod chemosensory sensilla facilitates fluid sampling

Abstract. Stomatopods, like many marine crustaceans, rely on their sense of smell to detect prey and to find mates (Ache 1982; Zimmer-Faust 1989; Atema & Voigt 1995). In lobsters, crabs, crayfishes, prawns, leptostracans, anaspidans, mysids, amphipods, tanaids, isopods, ostracodes, phyllopods, and cumaceans (Heimann 1984; Hallberg et al. 1992), this detection of odors from distant sources involves specialized chemosensory setae called aesthetascs located on the antennules. The external structure of stomatopod sensilla appears to follow the typical crustacean aesthetasc pattern, but their internal structure has not been previously examined. In this study, we use serial reconstruction from transmission electron microscopy to show that the stomatopod sensilla are aesthetascs. For chemoreception to occur, chemical-containing fluid must be very close to the surface of the aesthetascs, such that odor molecules can diffuse to chemoreceptors on the olfactory receptor neurons inside the aesthetasc. Flicking of stomatopod antennules maximizes fluid penetration near the parts of the sensilla where the cuticle is thinnest, and where the outer dendritic segments are most fully branched with the greatest surface area. Thus, the external and the internal structure of the stomatopod aesthetasc are "matched" to maximize the efficiency of odor arrival at the surface of the olfactory receptor neurons.     

Experimental and numerical determination of odorant solubility in nasal and olfactory mucosa

Odorant deposition in the nasal and olfactory mucosas is dependent on a number of factors including local air/odorant flow distribution patterns, odorant mucosal solubility and odorant diffusive transport in the mucosa. Although many of these factors are difficult to measure, mucosal solubility in the bullfrog mucus has been experimentally determined for a few odorants. In the present study an experimental procedure was combined with computational fluid dynamic (CFD) techniques to further describe some of the factors that govern odorant mucosal deposition. The fraction of odorant absorbed by the nasal mucosa (eta) was experimentally determined for a number of odorants by measuring the concentration drop between odorant 'blown' into one nostril and that exiting the contralateral nostril while the subject performed a velopharyngeal closure. Odorant concentrations were measured with a photoionization detector. Odorants were delivered to the nostrils at flow rates of 3.33 and 10 l/min. The velopharyngeal closure nasal air/odorant flows were then simulated using CFD techniques in a 3-D anatomically accurate human nose modeland the mucosal odorant uptake was numerically calculated. The comparison between the numerical simulations and the experimental results lead to an estimation of the human mucosal odorant solubility and the mucosal effective diffusive transport resistance. The results of the study suggest that the increase in diffusive resistance of the mucosal layer over that of a thin layer of water seemed to be general and non-odorant-specific; however, the mucosa solubility was odorant specific and usually followed the trend that odorants with lower water solubility were more soluble in the mucosa than would be predicted from water solubility alone. The ability of this approach to model odorant movement in the nasal cavity was evaluated by comparison of the model output with known values of odorant mucosa solubility.     

Broad activation of the glomerular layer enhances subsequent olfactory responses

Early olfactory experience with a specific odorant enhances the subsequent response of the glomerular layer of the rat olfactory bulb to that same odorant. Because different odorants activate different glomerular layer regions, it seemed plausible that experience with a large number of odorants might result in enhanced glomerular activation during subsequent exposure to both the previously experienced odorants and the novel odorants evoking activity in regions that overlapped with those previously stimulated by different odorants. To this end, 7 odorants were selected using our glomerular response data archive that together stimulated much of the glomerular layer (alpha-phellandrene, benzaldehyde, L-carvone, decanal, pentanol, santalol, and valeric acid). Young rats were exposed to a different odorant each day for 7 days, and this cycle was repeated 3 times from postnatal days 1-21. The [(14)C]2-deoxyglucose technique was used to measure neural activity in response to both previously experienced and novel odorants. The 2 novel odorants (alpha-ionone and L-menthone) activate regions of the glomerular layer that overlap with those stimulated by the 7 enrichment odorants. Our results indicate that early experience with multiple odorants results in increased responsiveness both to previously experienced odorants and to novel odorants that stimulate previously activated regions of the bulb.     

Numerical simulations of odorant detection by biologically inspired sensor arrays

The antennules of many marine crustaceans enable them to rapidly locate sources of odorant in turbulent environmental flows and may provide biological inspiration for engineered plume sampling systems. A substantial gap in knowledge concerns how the physical interaction between a sensing device and the chemical filaments forming a turbulent plume affects odorant detection and filters the information content of the plume. We modeled biological arrays of chemosensory hairs as infinite arrays of odorant flux-detecting cylinders and simulated the fluid flow around and odorant flux into the hair-like sensors as they intercepted a single odorant filament. As array geometry and sampling kinematics were varied, we quantified distortion of the flux time series relative to the spatial shape of the original odorant filament as well as flux metrics that may be important to both organisms and engineered systems attempting to measure plume structure and/or identify chemical composition. The most important predictor of signal distortion is the ratio of sensor diameter to odorant filament width. Achieving high peak properties (e.g. sharpness) of the flux time series and maximizing the total number of odorant molecules detected appear to be mutually exclusive design goals. Sensor arrays inspired specifically by the spiny lobster Panulirus argus and mantis shrimp Gonodactylaceus falcatus introduce little signal distortion but these species' neural systems may not be able to resolve plume structure at the level of individual filaments via temporal properties of the odorant flux. Current chemical sensors are similarly constrained. Our results suggest either that the spatial distribution of flux across the aesthetasc array is utilized by P. argus and G. falcatus, or that such high spatiotemporal resolution is unnecessary for effective plume tracking.     

Identification of Odorant-Receptor Interactions by Global Mapping of the Human Odorome

The human olfactory system recognizes a broad spectrum of odorants using approximately 400 different olfactory receptors (hORs). Although significant improvements of heterologous expression systems used to study interactions between ORs and odorant molecules have been made, screening the olfactory repertoire of hORs remains a tremendous challenge. We therefore developed a chemical systems level approach based on protein-protein association network to investigate novel hOR-odorant relationships. Using this new approach, we proposed and validated new bioactivities for odorant molecules and OR2W1, OR51E1 and OR5P3. As it remains largely unknown how human perception of odorants influence or prevent diseases, we also developed an odorant-protein matrix to explore global relationships between chemicals, biological targets and disease susceptibilities. We successfully experimentally demonstrated interactions between odorants and the cannabinoid receptor 1 (CB1) and the peroxisome proliferator-activated receptor gamma (PPARγ). Overall, these results illustrate the potential of integrative systems chemical biology to explore the impact of odorant molecules on human health, i.e. human odorome.     

Ultrastructure of aesthetasc innervation and external morphology of the lateral antennule setae of the spiny lobster Panulirus interruptus (Randall)

Summary Six types of setae and one type of cuticular depression were examined on the lateral antennule of the spiny lobster Panulirus interruptus using scanning electron microscopy. The organization and ultrastructure of the innervation of the most numerous setal type, the aesthetasc, were investigated using light-and transmission electron microscopy.Each aesthetasc is innervated by approximately 300 bipolar neurons whose sensory dendrites penetrate the hair and extend toward the tip, and whose axons project towards the central nervous system. The neuronal somata and two types of glia form a cluster within the antennular lumen. The inner sheath-cell somata encircle the dendritic tract distal to the sensory somata. These cells appear to extend distal processes which wrap the dendritic tract to the base of the aesthetasc. Elongate outer sheath cells are interposed between the glia-wrapped dendritic tract and the hypodermis which underlies the antennule cuticle. A continuous investment of neural lamella separates the hypodermis, the entire cluster of somata, and sensillar nerve from the antennule lumen. The organization of the neuronal somata and their association with outer and inner sheath cells in this marine species appear similar to those of crustaceans from freshwater and terrestrial habitats.