Variation of cleaning organ structures and setae of pedipalp tarsus in the family Phrynidae (Arachnida: Amblypygi).

The pedipalp tarsus of Amblypygi is used as a cleaning tool in grooming behavior and as a weapon in prey capture. The tarsus presents several structures with unknown functions that probably relate to both processes. The Amblypygi tarsus possesses a cleaning organ with two lines of projections and a group of setae distributed along the structure. We analyzed the morphological variation of the cleaning organ structures and the setae of the tarsus in species of the family Phrynidae using scanning electron microscopy (SEM). Additionally, we made histological sections to evaluate the sensory function of the structures. We found variation in the shape and size of the setae, projections and granular area of the cleaning organ; however, the observed differences do not allow for differentiation of taxonomic genera. The setae and projections of the cleaning organ have an internal structure similar to tip-pore sensilla. Externally, the setae are similar to the sensorial hairs in spiders and differ from the cleaning organ projections, which probably are modified setae. Mechanoreception function is evident in the setae, and is probable in projections of the cleaning organ; the evidence observed indicates that both structures may be considered as putative chemoreceptors.

Dorsal setae in Raoiella (Acari: Tenuipalpidae): Their functional morphology and implication in fluid secretion.

The setae of mites are not regarded as secretory structures, yet in the flat mite genus Raoiella, each developmental stage presents droplets of fluid associated with the tips of their dorsal setae. To understand the origin of this fluid, the ultrastructure of the dorsal setae is investigated in females of Raoiella bauchani Beard & Ochoa and the invasive pest species Raoiella indica Hirst using scanning and transmission electron microscopy techniques. The dorsal setae are barbed along their entire length and have either a broadened plumose or a flat spatulate tip. Ultrastructurally, they present the typical features of mechanoreceptors, but have a "hollow" axis represented by a protoplasmatic core containing dendritic branches. This combination of ultrastructural characters indicates that the setae might be multimodal receptors: acting as both mechanoreceptors and contact chemoreceptors. The epidermal cells that underlie the setal sockets are columnar and have an ultrastructure that suggests they have a glandular function. Moreover, these cells present regular microvilli apically and form extracellular cuticular canals, containing epicuticular filaments, that are connected with the microvilli proximally and which open via pores onto the surface of the setal base distally. This arrangement indicates that the secretion from the microvilli passes into the canals and is then conducted to pores at the base of the seta, where it then accumulates and moves up the setal shaft, along the longitudinal grooves of the barbs. Based on similar arrangements in some insect taxa, the organization of the structures here observed in Raoiella suggests the passage of a non-polar, water insoluble, lipoid fluid through the cuticle, the function of which is still obscure.

The structural basis for signal promiscuity in a bacterial chemoreceptor.

Signalling through chemosensory pathways is typically initiated by the binding of signal molecules to the chemoreceptor ligand binding domain (LBD). The PcaY_PP chemoreceptor from Pseudomonas putida KT2440 is characterized by an unusually broad signal range, and minimal requisites for signal binding are the presence of a C6-membered ring and that of a carboxyl group. Previous studies have shown that only some of the multiple signals recognized by this chemoreceptor are of apparent metabolic value. We report here high-resolution structures of PcaY_PP-LBD in the absence and presence of four cognate chemoeffectors and glycerol. The domain formed a four-helix bundle (4HB), and both ligand binding sites of the dimer were occupied with the high-affinity ligands protocatechuate and quinate, whereas the lower-affinity ligands benzoate and salicylate were present in only one site. Ligand binding was verified by microcalorimetric titration of site-directed mutants revealing important roles of an arginine and number of polar residues that establish an extensive hydrogen bonding network with bound ligands. The comparison of the apo and holo structures did not provide evidence for this receptor employing a transmembrane signalling mechanism that involves piston-like shifts of the final helix. Instead, ligand binding caused rigid-body scissoring movements of both monomers of the dimer. Comparisons with the 4HB domains of the Tar and Tsr chemoreceptors revealed significant structural differences. Importantly, the ligand binding site in PcaY_PP-LBD is approximately 8 Å removed from that of the Tar and Tsr receptors. Data indicate a significant amount of structural and functional diversity among 4HB domains. DATABASES: The coordinates and structure factors have been deposited in the protein data band with the following IDs: 6S1A (apo form), 6S18 (bound glycerol), 6S33 (bound protocatechuate), 6S38 (bound quinate), 6S3B (bound benzoate) and 6S37 (bound salicylate).

Diversity of Bacterial Chemosensory Arrays.

Chemotaxis is crucial for the survival of bacteria, and the signaling systems associated with it exhibit a high level of evolutionary conservation. The architecture of the chemosensory array and the signal transduction mechanisms have been extensively studied in Escherichia coli. More recent studies have revealed a vast diversity of the chemosensory system among bacteria. Unlike E. coli, some bacteria assemble more than one chemosensory array and respond to a broader spectrum of environmental and internal stimuli. These chemosensory arrays exhibit a great variability in terms of protein composition, cellular localization, and functional variability. Here, we present recent findings that emphasize the extent of diversity in chemosensory arrays and highlight the importance of studying chemosensory arrays in bacteria other than the common model organisms.

Masters of communication: The brain of the banded cleaner shrimp Stenopus hispidus (Olivier, 1811) with an emphasis on sensory processing areas.

The pan-tropic cleaner shrimp Stenopus hispidus (Crustacea, Stenopodidea) is famous for its specific cleaning behavior in association with client fish and an exclusively monogamous life-style. Cleaner shrimps feature a broad communicative repertoire, which is considered to depend on superb motor skills and the underlying mechanosensory circuits in combination with sensory organs. Their most prominent head appendages are the two pairs of very long biramous antennules and antennae, which are used both for attracting client fish and for intraspecific communication. Here, we studied the brain anatomy of several specimens of S. hispidus using histological sections, immunohistochemical labeling as well as X-ray microtomography in combination with 3D reconstructions. Furthermore, we investigated the morphology of antennules and antennae using fluorescence and scanning electron microscopy. Our analyses show that in addition to the complex organization of the multimodal processing centers, especially chemomechanosensory neuropils associated with the antennule and antenna are markedly pronounced when compared to the other neuropils of the central brain. We suggest that in their brains, three topographic maps are present corresponding to the sensory appendages. The brain areas which provide the neuronal substrate for these maps share distinct structural similarities to a unique extent in decapods, such as size and characteristic striated and perpendicular layering. We discuss our findings with respect to the sensory landscape within animal's habitat. In an evolutionary perspective, the cleaner shrimp's brain is an excellent example of how sensory potential and functional demands shape the architecture of primary chemomechanosensory processing areas.

Morphological and electrophysiological differences in tarsal chemosensilla between the wild silkmoth Bombyx mandarina and the domesticated species Bombyx mori.

Gustatory and olfactory senses of phytophagous insects play important roles in the recognition of host plants. In the domestic silkmoth Bombyx mori and its wild species Bombyx mandarina, the morphologies and responses of adult olfactory organs (antennae) have been intensely investigated. However, little is known about these features of adult gustatory organs and the influence of domestication on the gustatory sense. Here we revealed that both species have two types of sensilla (thick [T] and slim [S] types) on the fifth tarsomeres of the adult legs. In both species, females have 3.6-6.9 times more T-sensilla than males. Therefore, T-sensilla seem to play more important roles in females than in males. Moreover, gustatory cells of T-sensilla of B. mandarina females responded intensely to mulberry leaf extract in electrophysiological experiments, while T-sensilla of B. mori females (N4 strain) hardly responded to mulberry leaf extract. These results suggest that T-sensilla of B. mandarina females are involved in the recognition of oviposition sites. We also observed that, in three B. mori strains (N4, p50T, and Kinshu × Showa), the densities of sensilla on the fifth tarsomeres were much lower than in B. mandarina. These results indicate that domestication has influenced the tarsal gustatory system of B. mori.

Morphometric Analysis of Chemoreception Organ in Male and Female Ticks (Acari: Ixodidae).

The Haller's organ plays a crucial role in a tick's ability to detect hosts. Even though this sensory organ is vital to tick survival, the morphology of this organ is not well understood. The objective of this study was to characterize variation in the morphological components of the Haller's organ of three medically important tick species using quantitative methods. The Haller's organs of Ixodes scapularis Say (Ixodida: Ixodidae) (black-legged tick), Amblyomma americanum (L.) (Ixodida: Ixodidae) (lone star tick), and Dermacentor variabilis (Say) (Ixodida: Ixodidae) (American dog tick) were morphologically analyzed using environmental scanning electron microscopy and geometric morphometrics, and the results were statistically interpreted using canonical variate analysis. Our data reveal significant, quantitative differences in the morphology of the Haller's organ among all three tick species and that in D. variabilis the sensory structure is sexually dimorphic. Studies like this can serve as a quantitative basis for further studies on sensor physiology, behavior, and tick species life history, potentially leading to novel methods for the prevention of tick-borne disease.

The antennae of damselfly larvae.

The larval antennal sensilla of two Zygoptera species, Calopteryx haemorroidalis (Calopterygidae) and Ischnura elegans (Coenagrionidae) are investigated with SEM and TEM. These two species have different antennae (geniculate, setaceous) and live in different environments (lotic, lentic waters). Notwithstanding this, similarities in the kind and distribution of sensilla are outlined: in both species the majority of sensilla types is located on the apical portion of the antenna, namely a composed coeloconic sensillum (possible chemoreceptor), two other coeloconic sensilla (possible thermo-hygroreceptors) and an apical seta (direct contact mechanoreceptor). Other mechanoreceptors, such as filiform hairs sensitive to movements of the surrounding medium or bristles positioned to sense the movements of the flagellar segments, are present on the antenna. Similarities in the antennal sensilla types and distribution are observed also with other dragonfly species, such as Onychogomphus forcipatus and Libellula depressa. A peculiar structure with an internal organization similar to that of a gland is observed in the apical antenna of C. haemorroidalis and I. elegans and it is present also in O. forcipatus and L. depressa. The possible function of this structure is at the moment unknown but deserves further investigations owing to its widespread presence in Odonata larvae.

Progress and Potential of Electron Cryotomography as Illustrated by Its Application to Bacterial Chemoreceptor Arrays.

Electron cryotomography (ECT) can produce three-dimensional images of biological samples such as intact cells in a near-native, frozen-hydrated state to macromolecular resolution (∼4 nm). Because one of its first and most common applications has been to bacterial chemoreceptor arrays, ECT's contributions to this field illustrate well its past, present, and future. While X-ray crystallography and nuclear magnetic resonance spectroscopy have revealed the structures of nearly all the individual components of chemoreceptor arrays, ECT has revealed the mesoscale information about how the components are arranged within cells. Receptors assemble into a universally conserved 12-nm hexagonal lattice linked by CheA/CheW rings. Membrane-bound arrays are single layered; cytoplasmic arrays are double layered. Images of in vitro reconstitutions have led to a model of how arrays assemble, and images of native arrays in different states have shown that the conformational changes associated with signal transduction are subtle, constraining models of activation and system cooperativity. Phase plates, better detectors, and more stable stages promise even higher resolution and broader application in the near future.

Ultrastructure of chemoreceptive tarsal sensilla in an armored harvestman and evidence of olfaction across Laniatores (Arachnida, Opiliones).

Harvestmen (Arachnida, Opiliones) are especially dependent on chemical cues and are often regarded as animals that rely mainly on contact chemoreception. Information on harvestman sensilla is scarce when compared to other arachnid orders, especially concerning internal morphology. Using scanning (SEM) and transmission (TEM) electron microscopy, we investigated tarsal sensilla on the distal tarsomeres (DT) of all leg pairs in Heteromitobates discolor (Laniatores, Gonyleptidae). Furthermore, we explored the typological diversity of sensilla present on the DT I and II in members of the suborder Laniatores, which include two thirds of the formally described opilionid fauna, using species from 17 families representing all main laniatorian lineages. Our data revealed that DT I and II of H. discolor are equipped with wall-pored falciform hairs (two types), wall-pored sensilla chaetica (two types) and tip-pored sensilla chaetica, while DT III and IV are mainly covered with trichomes (non-sensory) and tip-pored sensilla chaetica. The ultrastructural characteristics support an olfactory function for all wall-pored sensilla and a dual gustatory/mechanoreceptive function for tip-pored sensilla chaetica. Based on our comparative SEM survey, we show that wall-pored sensilla occur in all investigated Laniatores, demonstrating their widespread occurrence in the suborder and highlighting the importance of both legs I and II as the sensory appendages of laniatorean harvestmen. Our results provide the first morphological evidence for olfactory receptors in Laniatores and suggest that olfaction is more important for harvestmen than previously thought.

Function of desiccate in gustatory sensilla of drosophila melanogaster.

Desiccate (Desi), initially discovered as a gene expressing in the epidermis of Drosophila larvae for protection from desiccation stress, was recently found to be robustly expressed in the adult labellum; however, the function, as well as precise expression sites, was unknown. Here, we found that Desi is expressed in two different types of non-neuronal cells of the labellum, the epidermis and thecogen accessory cells. Labellar Desi expression was significantly elevated under arid conditions, accompanied by an increase in water ingestion by adults. Desi overexpression also promoted water ingestion. In contrast, a knockdown of Desi expression reduced feeding as well as water ingestion due to a drastic decrease in the gustatory sensillar sensitivity for all tested tastants. These results indicate that Desi helps protect insects from desiccation damage by not only preventing dehydration through the integument but also accelerating water ingestion via elevated taste sensitivities of the sensilla.

Nano-architecture of gustatory chemosensory bristles and trachea in Drosophila wings.

In the Drosophila wing anterior margin, the dendrites of gustatory neurons occupy the interior of thin and long bristles that present tiny pores at their extremities. Many attempts to measure ligand-evoked currents in insect wing gustatory neurons have been unsuccessful for technical reasons. The functions of this gustatory activity therefore remain elusive and controversial. To advance our knowledge on this understudied tissue, we investigated the architecture of the wing chemosensory bristles and wing trachea using Raman spectroscopy and fluorescence microscopy. We hypothesized that the wing gustatory hair, an open-ended capillary tube, and the wing trachea constitute biological systems similar to nano-porous materials. We present evidence that argues in favour of the existence of a layer or a bubble of air beneath the pore inside the gustatory hair. We demonstrate that these hollow hairs and wing tracheal tubes fulfil conditions for which the physics of fluids applied to open-ended capillaries and porous materials are relevant. We also document that the wing gustatory hair and tracheal architectures are capable of trapping volatile molecules from the environment, which might increase the efficiency of their spatial detection by way of wing vibrations or during flight.

Spatiotemporal Coding of Individual Chemicals by the Gustatory System.

Four of the five major sensory systems (vision, olfaction, somatosensation, and audition) are thought to use different but partially overlapping sets of neurons to form unique representations of vast numbers of stimuli. The only exception is gustation, which is thought to represent only small numbers of basic taste categories. However, using new methods for delivering tastant chemicals and making electrophysiological recordings from the tractable gustatory system of the moth Manduca sexta, we found chemical-specific information is as follows: (1) initially encoded in the population of gustatory receptor neurons as broadly distributed spatiotemporal patterns of activity; (2) dramatically integrated and temporally transformed as it propagates to monosynaptically connected second-order neurons; and (3) observed in tastant-specific behavior. Our results are consistent with an emerging view of the gustatory system: rather than constructing basic taste categories, it uses a spatiotemporal population code to generate unique neural representations of individual tastant chemicals. SIGNIFICANCE STATEMENT: Our results provide a new view of taste processing. Using a new, relatively simple model system and a new set of techniques to deliver taste stimuli and to examine gustatory receptor neurons and their immediate followers, we found no evidence for labeled line connectivity, or basic taste categories such as sweet, salty, bitter, and sour. Rather, individual tastant chemicals are represented as patterns of spiking activity distributed across populations of receptor neurons. These representations are transformed substantially as multiple types of receptor neurons converge upon follower neurons, leading to a combinatorial coding format that uniquely, rapidly, and efficiently represents individual taste chemicals. Finally, we found that the information content of these neurons can drive tastant-specific behavior.

The antenna of a burrowing dragonfly larva, Onychogomphus forcipatus (Anisoptera, Gomphidae).

The larva of the dragonfly Onychogomphus forcipatus (Anisoptera, Gomphidae) has a burrowing lifestyle and antennae composed of four short and broad segments (scape, pedicel and a two-segmented flagellum). The present ultrastructural investigation revealed that different sensilla and one gland are located on the antenna. There is a great diversity of mechanoreceptors of different kinds. In particular club-shaped sensilla, sensilla chaetica, and tree-like sensilla show the typical structure of bristles, the most common type of mechanoreceptors, usually responding to direct touch, while numerous long thin thorny trichoid sensilla show a morphology recalling the structure of filiform hair mechanoreceptors. The latter ones are presumably important in larval Odonata for current detection and rheotactic orientation, especially in a burrowing species. On the smooth apical cuticle of the second flagellar segment, three structures are visible: (1) a small ellipsoidal pit hosting a convoluted peg, the morphology of which resembles that of a typical chemoreceptor (even if pores are lacking), (2) a couple of small pits (not investigated under TEM), and (3) one wide depression with spherical structures, the internal morphology of which lets us assume that it is a gland with unknown function. This is the first report of an antennal gland in palaeopteran insects.

Towards an Understanding of Molecule Capture by the Antennae of Male Beetles Belonging to the Genus Rhipicera (Coleoptera, Rhipiceridae).

Working on the hypothesis that an important function of the lamellate antennae of adult male beetles belonging to the genus Rhipicera is to detect scent associated with female conspecifics, and using field observations, anatomical models derived from X-ray microcomputed tomography, and scanning electron microscopy, we have investigated the behavioral, morphological, and morphometric factors that may influence molecule capture by these antennae. We found that male beetles fly upwind in a zigzag manner, or face upwind when perching, behavior consistent with an animal that is tracking scent. Furthermore, the ultrastructure of the male and female antennae, like their gross morphology, is sexually dimorphic, with male antennae possessing many more of a particular type of receptor-the sensillum placodeum-than their female counterparts (approximately 30,000 vs. 100 per antenna, respectively). Based on this disparity, we assume that the sensilla placodea on the male antennae are responsible for detecting scent associated with female Rhipicera beetles. Molecule capture by male antennae in their alert, fanned states is likely to be favoured by: (a) male beetles adopting prominent, upright positions on high points when searching for scent; (b) the partitioning of antennae into many small segments; (c) antennal morphometry (height, width, outline area, total surface area, leakiness, and narrow channels); (d) the location of the sensilla placodea where they are most likely to encounter odorant molecules; and (e) well dispersed sensilla placodea. The molecule-capturing ability of male Rhipicera antennae may be similar to that of the pectinate antennae of certain male moths.

Cholinergic epithelial cell with chemosensory traits in murine thymic medulla.

Specialized epithelial cells with a tuft of apical microvilli ("brush cells") sense luminal content and initiate protective reflexes in response to potentially harmful substances. They utilize the canonical taste transduction cascade to detect "bitter" substances such as bacterial quorum-sensing molecules. In the respiratory tract, most of these cells are cholinergic and are approached by cholinoceptive sensory nerve fibers. Utilizing two different reporter mouse strains for the expression of choline acetyltransferase (ChAT), we observed intense labeling of a subset of thymic medullary cells. ChAT expression was confirmed by in situ hybridization. These cells showed expression of villin, a brush cell marker protein, and ultrastructurally exhibited lateral microvilli. They did not express neuroendocrine (chromogranin A, PGP9.5) or thymocyte (CD3) markers but rather thymic epithelial (CK8, CK18) markers and were immunoreactive for components of the taste transduction cascade such as Gα-gustducin, transient receptor potential melastatin-like subtype 5 channel (TRPM5), and phospholipase Cß2. Reverse transcription and polymerase chain reaction confirmed the expression of Gα-gustducin, TRPM5, and phospholipase Cß2. Thymic "cholinergic chemosensory cells" were often in direct contact with medullary epithelial cells expressing the nicotinic acetylcholine receptor subunit α3. These cells have recently been identified as terminally differentiated epithelial cells (Hassall's corpuscle-like structures in mice). Contacts with nerve fibers (identified by PGP9.5 and CGRP antibodies), however, were not observed. Our data identify, in the thymus, a previously unrecognized presumptive chemosensitive cell that probably utilizes acetylcholine for paracrine signaling. This cell might participate in intrathymic infection-sensing mechanisms.

The epipharyngeal sensilla of the damselfly Ischnura elegans (Odonata, Coenagrionidae).

The knowledge on Odonata adult mouthparts sensilla is scanty and, notwithstanding the epipharynx in the labrum is considered an organ of taste, no ultrastructural investigation has been performed so far on this structure in Odonata. The labrum of the adult of the damselfly Ischnura elegans (Odonata, Coenagrionidae) shows on its ventral side the epipharynx with sensilla represented by articulated hairs and by small pegs located at the apex of slightly raised domes. Under scanning and transmission electron microscope, the articulated hairs, with a well developed socket and tubular body, have the typical structure of bristles, the most common type of insect mechanoreceptors, usually responding to direct touch; the pegs, showing an apical pore together with a variable number of sensory neurons (from two to five), the outer dendritic segments of which show a dendrite sheath stopping along their length, have features typical of contact chemoreceptors.

Distribution and innervation of putative arterial chemoreceptors in the bullfrog (Rana catesbeiana).

Peripheral arterial chemoreceptors have been located previously in the carotid labyrinth, the aortic arch, and the pulmocutaneous artery of frogs. In the present study we used cholera toxin B neuronal tract tracing and immunohistochemical markers for cholinergic cells (vesicular acetylcholine transporter [VAChT]), tyrosine hydroxylase (TH), and serotonin (5HT) to identify putative O2-sensing cells in Rana catesbeiana. We found potential O2-sensing cells in all three vascular areas innervated by branches of the vagus nerve, whereas only cells in the carotid labyrinth were innervated by the glossopharyngeal nerve. Cells containing either 5HT or TH were found in all three sites, whereas cells containing both neurotransmitters were found only in the carotid labyrinth. Cell bodies containing VAChT were not found at any site. The morphology and innervation of putative O2-sensing cells were similar to those of glomus cells found in other vertebrates. The presence of 5HT- and TH-immunoreactive cells in the aorta, pulmocutaneous artery, and carotid labyrinth appears to reflect a phylogenetic transition between the major neurotransmitter seen in the putative O2-sensing cells of fish (5HT) and those found in the glomus cells of mammals (acetylcholine, adenosine, and catecholamines).

How do mandibles sense? The sensory apparatus of larval mandibles in Palaemon elegans Rathke, 1837 (Decapoda, Palaemonidae).

The mandibles of decapod zoea-I larvae are robustly built masticating mouthparts equipped with several processes and spines. Superficial examination of these sturdy, inflexible structures can suggest that they are lacking sensory receptors. However, detailed TEM analysis of their ultrastructure revealed up to 11 sensillar cell clusters on the gnathal edges of the mandibles of the zoea-I in Palaemon elegans Rathke, 1837. Based on ultrastructural criteria we distinguish 7 types of sensilla: mechanoreceptors, chemoreceptors and mechano- and chemoreceptors. One sensory unit located at the base of the 'lacinia mobilis' exhibits the typical features of a crustacean mechanosensitive sensillum with an external seta and corresponding ultrastructure. Another unit shows features indicating bimodal contact chemosensitivity. A third one is similar to known olfactory chemoreceptors. Using the concept of modality-specific structures we analyse the structure and functional morphology of each sensillum, and give a comprehensive overview of the sensory abilities of zoea mandibles. We take a closer look at the ultrastructure of the 'lacinia mobilis', providing further features to trace its evolutionary history in Decapoda, and thus contributing to a better understanding of malacostracan phylogeny.