The chemosensilla on tarsi of Locusta migratoria (Orthoptera: Acrididae): Distribution,ultrastructure, expression of chemosensory proteins

The chemosensilla on the tarsi of Locusta migratoria were mapped using light microscopy, as well as scanning and transmission electron microscopy. Only chemosensilla chaetica were found on the tarsi. On the basis of their ultrastructure, these can be grouped into three main subtypes: short, long, and sunken sensilla chaetica. Short sensilla chaetica can be further divided into two groups containing 6 or 7 neurons. Long sensilla chaetica are mainly located on the lateral surface of the tarsi. Short sensilla chaetica were mainly found on the dorsal surface of the tarsi. Sunken sensilla chaetica were only found on the ventral surface, such as the pulvilli and arolium. Immunocytochemical localization of chemosensory protein (CSP) was performed on ultrathin sections of chemosensilla on tarsi. The antiserum against LmigCSP‐II intensively labeled all three types of sensilla chaetica. Gold granules were concentrated in the outer sensillum lymph surrounding the dendrite sheath, while the inner sensillum lymph containing dendrite branches was never labeled. Massive labeling with the anti‐LmigCSP‐II was also found in cuticle of the pulvilli on the ventral surface of tarsi. J. Morphol. 2009. © 2009 Wiley‐Liss, Inc.     

Intriguing similarities between two novel odorant-binding proteins of locusts

Two novel odorant-binding proteins (OBPs) of locust, LmigOBP2 and LmigOBP3 are very different from each other and from the previously reported LmigOBP1 in their amino acid sequences. Moreover, OBP3 contains three additional cysteines, a fact not previously recorded in standard length OBPs. However, these two proteins exhibit remarkably similar binding affinities to a set of organic compounds. Such behaviour is supported by three-dimensional models, showing very similar folding for LmigOBP2 and LmigOBP3, but clearly different for LmigOBP1. Also several amino acid residues lining the binding pockets of the three proteins appear conserved in LmigOBP2 and LmigOBP3, but not in LmigOBP1. Western blot experiments revealed the presence of LmigOBP2 in antennae, mouth parts and cerci, but could not detected LmigOBP3 in any of these tissues. In immunocytochemistry, antibodies against LmigOBP2 strongly stained the outer lymph of sensilla chaetica of the antennae, in contrast with LmigOBP1, previously reported in sensilla basiconica.     

Expression of odorant-binding and chemosensory proteins and spatial map of chemosensilla on labial palps of Locusta migratoria (Orthoptera: Acrididae)

The sensilla on labial palps in Locusta migratoria were observed and mapped using light microscopy, scanning and transmission electron microscopy. A dome region on the tip of the fourth segment (distal segment) of labial palps is mainly covered with sensilla chaetica (about 98%), and few sensilla basiconica (2%). The total number of both types of sensilla is significantly higher in females than in males. Sensilla chaetica can be further subdivided into three groups containing 6, 7 or 10 neurons. Immunocytochemical localization of odorant-binding protein (OBP) and chemosensory proteins (CSPs) was performed on ultrathin sections of sensilla on labial palps. The antiserum against odorant-binding protein from Locusta migratoria (LmigOBP) only labelled sensilla basiconica, with gold granules only found in the sensillum lymph. Chemosensory protein instead was specifically present in the outer sensillum lymph of all three subgroups of sensilla chaetica with antiserum against CSP-I from Schistocerca gregaria (SgreCSP-I). In contrast these three subgroups were never labelled with antiserum against CSP-II from Locusta migratoria (LmigCSP-II). In addition, a few sensilla chaetica could not be stained with any of the antisera used.     

Three odorant-binding proteins are co-expressed in sensilla trichodea of Drosophila melanogaster

Odorant-binding proteins (OBPs) are small soluble proteins present in the aqueous medium surrounding olfactory receptor neurones. In this study we examine the expression patterns of three Drosophila OBPs (LUSH=OBP76a, OS-E=OBP83b and OS-F=OBP83a), using post-embedding immunocytochemistry. All three OBPs are co-expressed in sensilla trichodea whereas sensilla intermedia show co-expression of OS-E and OS-F only, but not of LUSH. Thus, it is confirmed that an individual sensillum can contain more than one OBP, even if it comprises only a single receptor neurone, such as the subtype T-1. In s. trichodea of lush⁻ mutants, expression of OS-E and OS-F is not impaired. No other sensillum type on antenna or maxillary palp (e.g. sensilla basiconica, sensilla coeloconica) expresses LUSH, OS-E or OS-F. Within the s. trichodea the three OBPs show the same labelling pattern: the extracellular sensillum lymph in the hair lumen and the sensillum-lymph cavities are heavily labelled. Intracellularly, the three OBPs are co-localised in a variety of dense granules in all auxiliary cells, and also in the receptor neurones. Immunocytochemical data from antennal sections of flies where lush gene expression has been tagged with the reporter gene lacZ suggest that LUSH is synthesised only in the trichogen and the thecogen cells. Thus, LUSH OBP is produced and secreted by two auxiliary cells, whereas its turnover and decomposition does not appear to be restricted to these auxiliary cells but may also occur in the tormogen and receptor cells. The immunocytochemical results are discussed with respect to current concepts of the function of odorant-binding proteins.     

Expression of chemosensory proteins in hairs on wings of Locusta migratoria (Orthoptera: Acrididae)

The hairs on the wings of Locusta migratoria were observed and mapped using light microscopy, as well as by scanning and transmission electron microscopy. Based on their ultrastructure, we can distinguish four main types of hairs on the wings of adult L. migratoria , viz, short, medium and long hairs, and sensilla chaetica. The long hairs are located only on the ventral surface of the hindwing, whereas the other three types are present both on the dorsal and ventral surfaces of forewing and hindwing in both sexes. Medium hairs and sensilla chaetica are significantly more abundant on the dorsal surface of forewings in both females and males, than on the ventral surface, whereas the opposite was observed for short hairs (P < 0.01). No significant difference between males and females was observed in the density of any type of hairs (P > 0.1). Several dendritic branches, enveloped by a dendrite sheath, are situated in the lymph cavity of sensilla chaetica. Instead, no dendritic structure was observed in the cavity of the other three types of hairs. Immunocytochemical localization of chemosensory proteins (CSPs) was performed on ultrathin sections of hairs on wings. The antiserum against chemosensory proteins from L. migratoria (LmigCSP-II) strongly labelled sensilla chaetica, with gold granules only found in the outer sensillum lymph. In addition, the epidermal cell membrane of the wing was stained by the antiserum against LmigCSP-II. The other three types of hairs were never labelled. The results indicate that the wings might involve in contact chemoreception process.     

灰茶尺蠖成虫触角及幼虫头部感器超微结构

【目的】明确灰茶尺蠖Ectropis grisescens成虫触角及幼虫头部感器的种类、形态、数量和分布,以探讨灰茶尺蠖的行为机制。【方法】利用扫描电镜技术观察灰茶尺蠖雌、雄成虫触角和5龄幼虫头部感器的超微结构。【结果】灰茶尺蠖成虫触角上分布有8种感器,分别是栓锥形感器、耳形感器、毛形感器(Ⅰ-Ⅳ)、B?hm氏鬃毛、腔锥形感器(Ⅰ和Ⅱ)、鳞形感器、锥形感器(Ⅰ和Ⅱ)和刺形感器。其中,栓锥形感器仅分布在雌蛾触角上,耳形感器、毛形感器(STⅠ-Ⅲ)仅分布在雄虫触角上。5龄幼虫触角上着生1个栓锥形感器、1个锥形感器和2个刺形感器;上唇着生有6对刺形感器,内唇着生有3对刺形感器和1对指形感器;上颚基部外侧着生有2个刺形感器;下颚及下颚须着生有5个刺形感器、9个锥形感器和2个栓锥形感器;下唇须着生有1个锥形感器和1个刺形感器;吐丝器前端着生有1对刺形感器。【结论】灰茶尺蠖雌、雄成虫触角感器存在性二型性,且雄虫上感器种类和数量较多,据此推测雄虫感受寄主植物或性信息素的能力较强;幼虫头部感器具有嗅觉和味觉功能,在其判断食物的种类和适应性等生态行为中发挥重要作用。     

Expression pattern analysis of odorant‐binding proteins in the pea aphid Acyrthosiphon pisum

Odorant‐binding proteins (OBPs) are soluble proteins mediating chemoreception in insects. In previous research, we investigated the molecular mechanisms adopted by aphids to detect the alarm pheromone (E)‐β‐farnesene and we found that the recognition of this and structurally related molecules is mediated by OBP3 and OBP7. Here, we show the differential expression patterns of 5 selected OBPs (OBP1, OBP3, OBP6, OBP7, OBP8) obtained performing quantitative RT‐PCR and immunolocalization experiments in different body parts of adults and in the 5 developmental instars, including winged and unwinged morphs, of the pea aphid Acyrthosiphon pisum. The results provide an overall picture that allows us to speculate on the relationship between the differential expression of OBPs and their putative function. The expression of OBP3, OBP6, and OBP7 in the antennal sensilla suggests a chemosensory function for these proteins, whereas the constant expression level of OBP8 in all instars could suggest a conserved role. Moreover, OBP1 and OBP3 are also expressed in nonsensory organs. A light and scanning electron microscopy study of sensilla on different body parts of aphid, in particular antennae, legs, mouthparts, and cornicles‐cauda, completes this research providing a guide to facilitate the mapping of OBP expression profiles.     

悬铃木方翅网蝽触角感器扫描电镜观察

利用扫描电镜对悬铃木方翅网蝽Corythucha ciliata(Say)雌、雄成虫触角背面和腹面进行观察。结果表明:悬铃木方翅网蝽触角为棒状,共4节,分为柄节、梗节和鞭节。触角上共有4种感器,分别为刺型感器、锥形感器、毛型感器和芽型感器;这些感器不存在性二型现象。其中,刺型感器分为大刺型感器和小刺型感器2种类型;芽型感器首次在异翅亚目昆虫触角上发现。雄成虫触角感器数量明显多于雌成虫,不同类型的感器在触角各节上的数量与分布各不相同。     

Reproductive strategies and parasitization behavior of Ageniaspis citricola, a parasitoid of the citrus leafminer Phyllocnistis citrella

We describe the number, distribution, and function of sensilla located on different organs of Lobesia botrana (Lepidoptera: Tortricidae) females using scanning electron microscopy, selective staining, and contact electrophysiology. The tarsi of the prothoracic legs bear contact chemo‐mechanoreceptor sensilla chaetica (5–13 per tarsomere), arranged in rings mainly concentrated on ventral surfaces, and different mechanosensory structures (sensilla chaetica, sensilla squamiformia, sensilla campaniformia, and spines). A single contact chemo‐mechanoreceptor sensillum chaeticum is present between the claws on the pretarsus. The ventral surface of the ovipositor lobes is covered with numerous mechanosensory sensilla chaetica of different types, out of which 10 have a contact chemosensory function. Putative contact chemo‐mechanoreceptor sensilla were also observed on the proboscis and antenna. Longitudinal rows of alternated sensilla styloconica and basiconica are present on the distal part of the proboscis, and rings of sensilla chaetica are present at the antennal tip. The sensilla on these body parts may play different roles in the selection of an oviposition site.     

Ultrastructural characterization of olfactory sensilla and immunolocalization of odorant binding and chemosensory proteins from an ectoparasitoid Scleroderma guani (Hymenoptera: Bethylidae)

The three-dimensional structures of two odorant binding proteins (OBPs) and one chemosensory protein (CSP) from a polyphagous ectoparasitoid Scleroderma guani (Hymenoptera: Bethylidae) were resolved bioinformatically. The results show that both SguaOBP1 and OBP2 are classic OBPs, whereas SguaCSP1 belongs to non-classic CSPs which are considered as the "Plus-C" CSP in this report. The structural differences between the two OBPs and between OBP and CSP are thoroughly described, and the structural and functional significance of the divergent C-terminal regions (e.g., the prolonged C-terminal region in SguaOBP2 and the additional pair of cysteines in SguaCSP1) are discussed. The immunoblot analyses with antisera raised against recombinant SguaOBP1, OBP2, and CSP1, respectively, indicate that two SguaOBPs are specific to antennae, whereas SguaCSP1, which are more abundant than OBPs and detected in both male and female wasps, expresses ubiquitously across different tissues.We also describe the ultrastructure of the antennal sensilla types in S. guani and compare them to 19 species of parasitic Hymenoptera. There are 11 types of sensilla in the flagellum and pedicel segments of antennae in both male and female wasps. Seven of them, including sensilla placodea (SP), long sensilla basiconica (LSB), sensilla coeloconica (SC), two types of double-walled wall pore sensilla (DWPS-I and DWPS-II), and two types of sensilla trichodea (ST-I and ST-II), are multiporous chemosensilla. The ultralsturctures of these sensilla are morphologically characterized. In comparison to monophagous specialists, the highly polyphagous generalist ectoparasitoids such as S. guani possess more diverse sensilla types which are likely related to their broad host ranges and complex life styles. Our immunocytochemistry study demonstrated that each of the seven sensilla immunoreacts with at least one antiserum against SguaOBP1, OBP2, and CSP1, respectively. Anti-OBP2 is specifically labeled in DWPS-II, whereas the anti-OBP1 shows a broad spectrum of immunoactivity toward four different sensilla (LSB, SP, ST-I and ST-II). On the other hand, anti-CSP1 is immunoactive toward SP, DWPS-I and SC. Interestingly, a cross co-localization pattern between SguaOBP1 and CSP1 is documented for the first time. Given that the numbers of OBPs and CSPs in many insect species greatly outnumber their antennal sensilla types, it is germane to suggest such phenomenon could be the rule rather than the exception.     

The antennae of neopseustid moths: Morphology and phylogenetic implications, with special reference to the sensilla (Insecta, Lepidoptera, Neopseustidae)

The antennae of Lepidoptera Neopseustidae were examined with the scanning electron microscope. The studied species, Nematocentropus cfr. omeiensis, Neopseustis meyricki, Synempora andesae, Apoplania valdiviana and Apoplania penai possess nine types of antennal flagellum sensilla: multiporous large sensilla basiconica, multiporous thin sensilla basiconica, multiporous small sensilla basiconica, multiporous sensilla trichodea, multiporous sensilla coeloconica; uniporous sensilla chaetica; aporous sensilla chaetica, aporous stylus-shaped sensilla chaetica, aporous sensilla styloconica.The multiporous sensillum type here termed “multiporous large sensillum basiconicum” is unknown from other Lepidoptera and probably constitutes an autapomorphy of the family Neopseustidae. This sensillum type is remarkable by having a single base in female Apoplania and Synempora while in male Apoplania it has a bifid or trifid base, and in male Synempora it is composed of two or three incompletely separated hairs. This may be the first recorded example of a sexually dimorphic lepidopteran sensillum type. The stylus-shaped sensillum chaeticum is a primitive type which occurs only in some lower Lepidoptera.     

Structure and sensilla of the antennae and mouthparts of Loxocephala perpunctata Jacobi (Hemiptera: Fulgomorpha: Eurybrachidae)

The ultramorphology of the antennae and mouthparts of the adult Loxocephala perpunctata Jacobi was studied through a scanning electron microscope. Seven types of sensilla were found on antennomeres, including a Böhm bristle on the scape, sensillum trichoideum and plaque organ on the pedicel, two subtypes of sensilla chaetica and two subtypes of sensilla campaniformia on these two antennomeres; and Bourgoin's organ with sensory pegs and sensilla basiconicum on the basal bulb of the flagellum. The mouthparts of L. perpunctata are of the typical piercing-sucking type, similar to mouthparts found in other hemipteran insects. In general, six types of sensilla (i.e., four subtypes of sensilla chaetica, sensillum basiconicum, subapical labial sensillum, uniporous peg-like sensillum, multiporous peg-like sensillum and two subtypes of bristle-like sensilla) were detected on different locations of the labium, with the last three, and numerous cuticular processes, present on the labial tip. The potential functions of these sensilla are discussed.     

Expression patterns of odorant-binding proteins in antennae of the moth<Emphasis Type="Italic"> Manduca sexta</Emphasis>

Monoclonal antibodies (MAbs) were generated to six recombinant proteins (odorant-binding proteins; OBPs) of Manduca sexta. The specificity of each MAb was demonstrated by labeling six immunoblots, each of which contained samples of all six recombinant OBPs. The expression patterns of the six OBPs could be grouped into three classes: (1) one (GOBP1) was expressed in sensilla located throughout each annulus; (2) two (ABPX and ABP2) were expressed in the long sensilla trichoidea bordering a zone that was arranged as an arch on the periphery of each annulus; (3) three (PBP2, PBP3, and GOBP2) were expressed in shorter sensilla occupying a wedge-shaped mid-annular zone of each annulus. In female antennae, sensilla expressing these OBPs were intermixed, and the distinct zonation observed in the male antenna was absent. In males, PBP2 was co-expressed in exactly the same cells of the mid-annular zone as those expressing PBP3 and most of the same cells expressing GOBP2, although its expression overlapped with no or only a few sensilla expressing OBPs of class 1 (GOBP1) or class 2 (ABPX, ABP2). This overlap of expression or lack of overlap between PBP2 and the other OBPs for male antennae was mirrored in female antennae. In view of the restricted spatial expression of OBPs within an annulus and the diversity of possible dimeric combinations of OBPs that arises from the co-expression of multiple OBPs in a given sensillum, OBPs could contribute to the specificity of the olfactory responses of insects.This research was supported by grants from the National Science Foundation (IBN-9604095) and the University of Illinois Critical Research Initiatives     

A large family of divergent Drosophila odorant-binding proteins expressed in gustatory and olfactory sensilla.

We identified a large family of putative odorant-binding protein (OBP) genes in the genome of Drosophila melanogaster. Some of these genes are present in large clusters in the genome. Most members are expressed in various taste organs, including gustatory sensilla in the labellum, the pharyngeal labral sense organ, dorsal and ventral cibarial organs, as well as taste bristles located on the wings and tarsi. Some of the gustatory OBPs are expressed exclusively in taste organs, but most are expressed in both olfactory and gustatory sensilla. Multiple binding proteins can be coexpressed in the same gustatory sensillum. Cells in the tarsi that express OBPs are required for normal chemosensation mediated through the leg, as ablation of these cells dramatically reduces the sensitivity of the proboscis extension reflex to sucrose. Finally, we show that OBP genes expressed in the pharyngeal taste sensilla are still expressed in the poxneuro genetic background while OBPs expressed in the labellum are not. These findings support a broad role for members of the OBP family in gustation and olfaction and suggest that poxneuro is required for cell fate determination of labellar but not pharyngeal taste organs.     

Differential expression of odorant-binding proteins in the mandibular glands of the honey bee according to caste and age

Odorant-binding proteins (OBPs) and chemosensory proteins (CSPs) mediate both perception and release of chemical stimuli in insects. The genome of the honey bee contains 21 genes encoding OBPs and 6 encoding CSPs. Using a proteomic approach, we have investigated the expression of OBPs and CSPs in the mandibular glands of adult honey bees in relation to caste and age. OBP13 is mostly expressed in young individuals and in virgin queens, while OBP21 is abundant in older bees and is prevalent in mated queens. OBP14, which had been found in larvae, is produced in hive workers' glands. Quite unexpectedly, the mandibular glands of drones also contain OBPs, mainly OBP18 and OBP21. We have expressed three of the most represented OBPs and studied their binding properties. OBP13 binds with good specificity oleic acid and some structurally related compounds, OBP14 is better tuned to monoterpenoid structures, while OBP21 binds the main components of queen mandibular pheromone as well as farnesol, a compound used as a trail pheromone in the honey bee and other hymenopterans. The high expression of different OBPs in the mandibular glands suggests that such proteins could be involved in solubilization and release of semiochemicals.     

绿盲蝽触角感器的扫描电镜观察

利用扫描电镜对绿盲蝽雌雄成虫的触角形态和感器进行了观察。结果表明： 绿盲蝽触角为线形, 共4节, 即柄节、梗节和2个鞭节。触角感器共有4种, 分别为毛形感器、刺形感器、锥形感器、Böhm氏鬃毛。其中, 毛形、刺形、锥形感器各有两种类型。雌雄个体之间触角感器的类型、分布均没有明显差异。不同感器在触角各节上的数量与分布各不同。     

Molecular and functional characterization of three odorant binding proteins from the oriental fruit moth Grapholita molesta (Busck) (Lepidoptera: Tortricide)

Odorant binding proteins (OBPs) act in recognizing odor molecules and their most well‐studied functions are transporting odors across the sensillum lymph to olfactory receptor neurons within the insect antennal sensillum. The adults of Grapholita molesta highly depend on olfactory cues in locating host plants and selecting oviposition sites, in which OBPs play an important role in perceiving and recognizing host plant volatiles. Exploring the physiological function of OBPs could facilitate our understanding of their importance in insects’ chemical communication. In this study, three OBP genes were cloned and named GmolOBP4, GmolOBP5, and GmolOBP10. Quantitative real‐time PCR results indicated that GmolOBP4 and GmolOBP10 were predominantly expressed in adult antennae and GmolOBP5 was expressed in multiple tissues, including head, legs, and wings in addition to antennae. The binding affinities of the three recombinant GmolOBPs (rGmolOBPs) with four sex pheromone components and twenty‐nine host plant volatiles were measured using 1‐N‐Phenyl‐naphthylamine as a fluorescence probe. The three rGmolOBPs exhibited specific binding properties to potential ligands, GmolOBP4 and GmolOBP10 bound to minor sex pheromone components, such as (Z)‐8‐dodecenyl alcohol and dodecanol, respectively. rGmolOBP4 showed intermediate binding ability with hexanal, benzyl alcohol, and pear ester, rGmolOBP5 had a weak affinity for benzaldehyde, pear ester and, methyl jasmonate, and rGmolOBP10 showed strong binding capacity toward hexanol, decanol, and α‐ocimene. We speculate that the GmolOBP4 and GmolOBP10 have dual functions in perception and recognition of host plant volatiles and sex pheromone components, while GmolOBP5 may serve other function(s).