Trichogen Cells Research Articles

Nanopore Formation in the Cuticle of an Insect Olfactory Sensillum

Nanometer-level patterned surface structures form the basis of biological functions, including superhydrophobicity, structural coloration, and light absorption [1-3]. In insects, the cuticle overlying the olfactory sensilla has multiple small (50- to 200-nm diameter) pores [4-8], which are supposed to function as a filter that admits odorant molecules, while preventing the entry of larger airborne particles andlimiting water loss. However, the cellular processes underlying the patterning of extracellular matrices into functional nano-structures remain unknown. Here, we show that cuticular nanopores in Drosophila olfactory sensilla originate from a curved ultrathin film that is formed in the outermost envelope layer of the cuticle and secreted from specialized protrusions in the plasma membrane of the hair forming (trichogen) cell. The envelope curvature coincides with plasma membrane undulations associated with endocytic structures. The gore-tex/Osiris23 gene encodes an endosomal protein that is essential for envelope curvature, nanopore formation, and odor receptivity and is expressed specifically in developing olfactory trichogen cells. The 24-member Osiris gene family is expressed in cuticle-secreting cells and is found only in insect genomes. These results reveal an essential requirement for nanopores for odor reception and identify Osiris genes as a platform for investigating the evolution of surface nano-fabrication in insects.

Atypical mechanoreceptors in larvae of biting midges Forcipomyia nigra (Diptera: Ceratopogonidae)

The dorsolateral setae of Forcipomyia nigra have been investigated. They are mechanoreceptors, sensilla trichoidea, innerved by a single neuron. The tubular body is located at the base of the hair shaft. Although the fine structure of the dorsolateral setae is similar to dorsal setae that perform a double function: secretion of a hygroscopic viscous substance through the pores and mechanoreception dorsolateral setae, they do not secrete any fluid. In both types of setae, trichogen cells (hair forming cells) produce the hair shafts and thereafter do not retract from the cavity of the setae. They contain a large polyploid nucleus and expanded bundles of microtubules. In dorsolateral mechanoreceptors, the microtubules form a network around the nucleus of the trichogen cell and are especially numerous in the cytoplasm invading the interior of the seta, which is evidenced by immunofluorescence light microscopy. No tormogen cell, responsible for the production of the setal socket, was found. Our observations indicate that the dorsolateral setae are solely mechanoreceptors but their trichogen cells reveal some glandular activity.

Ultrastructure of mediodorsal setae in biting midge larvae of the genus Atrichopogon Kieffer with notes on their biological significance (Diptera: Ceratopogonidae)

The ultrastructure of the strong mediodorsal setae in terrestrial stage IV larvae of Atrichopogon (Meloehelea) oedemerarum and A. (M.) meloesugans was examined using light, scanning and transmission electron microscopy. Serrated setae placed on prominent processes are distributed in pairs on all thoracic and abdominal segments. Setae are innervated by a single dendrite and their surface has no pores. The trichogen cell is not retracted from the setal lumen on completion of the hair-forming process but fills the mediodorsal seta also when the larval cuticle is fully sclerotised. Such a phenomenon was previously reported in terrestrial larvae of the genus Forcipomyia. We suggest that the mediodorsal setae described in Atrichopogon are plesiotypic mechanoreceptors for the subfamily Forcipomyiinae. They are preserved in the truly terrestrial larvae of Atrichopogon, but modified to secretory setae in the genus Forcipomyia. Both genera bearing distinct mediodorsal setae have developed functional tracheal gills, unknown in other biting midges.

Morphology and histology of secretory setae in terrestrial larvae of biting midges of the genus Forcipomyia (Diptera: Ceratopogonidae)

Apneustic larvae of the genus Forcipomyia possess unique secretory setae located on the dorsal surface along the body in two rows, one pair on each thoracic and abdominal segment and two pairs on the head. Morphological and histological studies of secretory setae in fourth instar larvae of Forcipomyia nigra (Winnertz) and Forcipomyia nigrans Remm indicate they are modified mechanoreceptors (sensilla trichodea) in which the trichogen cell is a glandular cell producing a hygroscopic secretion. The cytoplasm of the glandular trichogen cell fills the lumen of a secretory seta, which shows one or more pores on the apex. The cytoplasm contains numerous microtubules responsible for transportation of proteinaceous vesicles, and an extremely large polyploid nucleus typical of gland cells. The main role of the hygroscopic secretion is to moist the body and thus facilitate cuticular respiration.

Discrete regulatory regions control early and late expression of D‐Pax2 during external sensory organ development

The transcription factor D-Pax2 is required for the correct differentiation of several cell types in Drosophila sensory systems. While the regulation of its expression in the developing eye has been well studied, little is known about the mechanisms by which the dynamic pattern of D-Pax2 expression in the external sensory organs is achieved. Here we demonstrate that early activation of D-Pax2 in the sensory organ lineage and its maintenance in the trichogen and thecogen cells are governed by separate enhancers. Furthermore, the initial activation is controlled in part by proneural proteins whereas the later maintenance expression is regulated by a positive feedback loop.

Developmental expression of odorant‐binding proteins and chemosensory proteins in the embryos of Locusta migratoria

We have investigated the development of chemosensilla and the secretion of odorant-binding proteins (OBPs) and chemosensory proteins (CSPs) in the embryo of Locusta migratoria manilensis. We first report the changes of each sensillum in embryo just preceding hatch in detail and show that different sensilla have different developmental processes. Trichogen cells are first involved in forming the structure of pegs, and then, after retraction, they start secreting OBPs and CSPs in the sensillar lymph. The synthesis of LmigOBP1 starts during the embryogenesis about 0.5 h preceding hatching, specifically in sensilla trichodea and basiconica of the antenna. LmigOBP2, instead, was only found in the outer sensillum lymph (oSl) of sensilla chaetica of the antenna, while we could not detect LmigOBP3 in any type of sensilla of the antenna. The ontogenesis of CSPs in the embryos is similar to that of OBPs. Expression of CSPI homolog in Locusta migratoria is detected using the antiserum raised against SgreCSPI. CSPI is specifically expressed in the outer sensillum lymph of sensilla chaetica of the antenna, and anti-LmigCSPII dose not label any sensilla of the embryos. These data indicate that in locusts, OBPs and CSPs follow different temporal expression patterns, and also that OBPs are expressed in different types of sensilla.

Photographic map of the polytene chromosomes of Cochliomyia hominivorax

Cochliomyia hominivorax (Coquerel) (Diptera: Calliphoridae) is one of the most important myiasis-causing flies and is responsible for severe economic losses to the livestock industry throughout the Neotropical region. A polytene chromosome map is an invaluable tool for the genetic analysis and manipulation of any species because it allows the integration of physical and genetic maps. Cochliomyia hominivorax has a diploid number of 12 chromosomes (2n = 12): five pairs of autosomes and one pair of sex chromosomes (XX/XY), which do not polytenize. We created a new photomap of the polytene chromosomes of C. hominivorax describing its five autosomes (chromosomes 2-6). Pupal trichogen cells, which have chromosomes with a high degree of polytenization, were used to elaborate this map. The photomap was made by comparing 20 different nuclei and choosing, for each chromosome segment, the region with the highest resolution. Thus, we present a new photomap of the five autosomes of this species, with a total resolution of 1450 bands.

Polytene chromosome maps in four species of tsetse flies Glossina austeni, G. pallidipes, G. morsitans morsitans and G. m. submorsitans (Diptera: Glossinidae): a comparative analysis

Photographic polytene chromosome maps from pupal trichogen cells of four tsetse species, Glossina austeni, G. pallidipes, G. morsitans morsitans and G. m. submorsitans were constructed and compared. The homology of chromosomal elements between the species was achieved by comparing banding patterns. The telomeric and subtelomeric chromosome regions were found to be identical in all species. The pericentromeric regions were found to be similar in the X chromosome and the left arm of L1 chromosome (L1L) but different in L2 chromosome and the right arm of L1 chromosome (L1R). The L2 chromosome differs by a pericentric inversion that is fixed in the three species, G. pallidipes, G. morsitans morsitans and G. m. submorsitans. Moreover, the two morsitans subspecies appeared to be homosequential and differ only by two paracentric inversions on XL and L2L arm. Although a degree of similarity was observed across the homologous chromosomes in the four species, the relative position of specific chromosome regions was different due to chromosome inversions established during their phylogeny. However, there are regions that show no apparent homology between the species, an observation that may be attributed to the considerable intra--chromosomal rearrangements that have occurred following the species divergence. The results of this comparative analysis support the current phylogenetic relationships of the genus Glossina.

Scale length/wing length correlation in Lepidoptera (Insecta)

Significant positive correlation between wing length and scale length was found in a broad sample of lepidopteran taxa. No breakdown of the correlation was observed in the lowest wing size range. The best fit is a non-linear (power) curve. The minimum individual scale length observed was 40 µm (in a nepticulid), while the maximum was close to 500 µm (in a castniid; there are literature records of even larger scales in this family). Scale size variation probably reflects variation in the size of scale-forming trichogen cells. The latter, in turn, may at least partly reflect difference in genome size, such as has long been known to be the case with the cells that form, respectively, the cover and ground scales in some higher moths.

Cellular organization and fine structure of type II microtrich sensilla in gammaridean amphipods (Crustacea)

The cellular organization of type II microtrich sensilla was studied in male Anonyx lilljeborgi Boeck, 1871 (Lysianassoidea) by light and transmission electron microscopy. The sensillum consists of two bipolar sensory neurons and three concentric sheath cells. The sensory cell bodies are subepidermal. In each sensillum both dendrites are enclosed by the thecogen cell process. The inner dendritic segments are filled with mitochondria and lucent vesicles and expand in the epidermis into a spindle-shaped swelling. One of the neurons gives rise to two cilia and the second to a single cilium. These three outer dendritic segments lie in the receptorlymph cavity. The dendritic sheath, secreted by the thecogen cell process, completely ensheaths the outer dendritic segments. The trichogen (middle) cell and the tormogen (outer) cell incompletely enclose the thecogen cell, but their processes form autojunctions around the dendritic sheath in the apical epidermis. In premolt, the trichogen cell processes project into the exuvial space. The cytoplasm of the tormogen cell and the bordering epidermal cells contains coarse osmiophilic inclusions. All the cells of the sensillum are joined by desmosomes. The sensilla structurally resemble chemosensory (gustatory) insect sensilla.

Developmental expression of Ultraspiracle proteins in the epidermis of the tobacco hornworm, Manduca sexta, during larval life and the onset of metamorphosis.

During the final two larval instars, a changing pattern of three Ultraspiracle (Usp) proteins (50.5, 52.5, and 57kDa) was detected in immunoblots of the dorsal abdominal epidermis of the tobacco hornworm, Manduca sexta, by a monoclonal antibody against Drosophila Usp that was shown to detect MsUsp. The 57- and 52.5-kDa bands were present during the intermolt periods and the 50.5- and 52.5-kDa bands during the molting phases. The antibody detected a nuclear antigen present in epidermis, muscle, fat body, and the central nervous system from the time of hatching. In the epidermis Usp was present in all cell nuclei but was especially prominent in the tormogen and trichogen cells immediately after ecdysis in both the penultimate and final instars. This latter immunoreactivity disappeared within 12h whereas the remainder of the epidermis retained high levels throughout the feeding period. During the molt immunostaining reappeared in the hair cell nuclei. During the wandering stage at the onset of metamorphosis and just before pupal ecdysis, immunoblots showed high levels of Usp, but nuclei showed little or no staining. This discrepancy is likely due to the loss of one Usp isoform from the nucleus and its dispersal in the cytoplasm in preparation for the appearance of the second isoform.

Primary culture of antennal cells of Mamestra brassicae: morphology of cell types and evidence for biosynthesis of pheromone-binding proteins in vitro.

A culture technique for the in vitro growth and differentiation of antennal cells of Mamestra brassicae is described. The morphology of the growing cells is described. At least five morphological cell types can be distinguished in the cultures. Neuronal cells (type A) were characterized morphologically and immunocytochemically with anti-HRP staining and could be separated into two subtypes, A1 and A2, on the basis of the size of the cell body. Non-neuronal cells could be divided into four types. Type-B cells consisted of large and flat cells with a veil-like cytoplasm. They usually adhered to each other and formed groups of 20 to 50 cells. Type C were large and flat cells, highly variable in size and shape. They closely resembled insect glial cells in culture. Type-D cells grew thick processes, up to 100 microm long. These elongated processes resembled trichoid hairs, and D cells are most probably trichogen cells. Type E were spindle-shaped cells present in low number in the cultures. A high proportion of similar cells in the hemolymph of pupae suggests that E cells are hemocytes. Pheromone-binding proteins were detected using specific antisera in the medium of 3- to 4-week-old cultures.

Fine Structure of the Putative Pheromone-Sensitive Sensilla on the Antennae of Andraca bipunctata (Lepidoptera: Bombycidae)

The antennae of Andraca bipunctata Walker are sexually dimorphic; those of males are bipectinate with 43 pairs of branches, and those of females are filiform without branches. In both sexes, most sensilla are on the frontal (windward, ventral) side, but the types of sensilla differ in the 2 sexes. The main type in males is sensillum trichodeum (s. trichodeum), and in females are s. basiconicum and the short s. chaeticum. Each putative pheromone-sensitive sensillum, trichodeum, is innervated by 2 bipolar neurons containing many mitochondria, multivesicular bodies, rough endoplasmic reticulum, microtubles, and lamellar bodies. The dendrite is subdivided into inner and outer dendritic segments between the ciliary region, which is characterized by proximal and distal bodies. The outer dendritic segment, which contains microtubules, penetrates the cuticular canal and extends into the hair shaft. Outside the neurons, thecogen, trichogen, and tormogen cells surround the neural bodies, forming the inner, intermediate, and outer sheath below the hair shaft. This s. trichodeum has multipores and each pore has 2-3 pore tubules within the pore canal. These pore tubules extend into the hair lumen and probably help the pheromone molecules contact the dendritic membrane receptor, because most pheromones are water insoluble.

Structure and development of larval antennae in embryos ofLytta viridanaLeConte (Coleoptera: Meloidae)

At time of hatch (252–264 h at 25 ± 0.5 °C), each antenna in Lytta viridana has three flagellomeres, three extrinsic muscles, and 25 sensilla of five different types, including a large composite sensillum of 19 sensory units on flagellomere II. Each antenna evaginates from epidermis on either side the stomodaeum beginning at 16% of embryogenesis. At 21%, a cell near its apex divides into two pioneer neurons that move into its lumen and project their axons to the brain by 29%. Sensillar stem cells begin to emerge at 23%, those of the appendix within a large embryonic placode and, from 26 to 48%, divide asymmetrically to generate the neurons and accessory cells of each sensillum. Sensillar axonogenesis begins at 34%, the first axons contact the brain at 35%, and antennal glomeruli begin to form within the deutocerebra at 57%. At 35%, the trichogen cell of each sensillum begins to grow out and larval cuticle is deposited about these, beginning at 57%. Upon withdrawal of trichogen cytoplasm from within the appendix at 81%, the dendrites of each sensory unit grow into it and begin to branch. Functional aspects are addressed and the observations compared with the limited information available on embryos of other insects.

Fine Structure of the Trichobothria and their Regeneration during Moulting in the Whip Scorpion Typopeltis crucifer Pocock, 1894

Abstract The highly complicated structure of the trichobothrium in the whip scorpion Typopeltis crucifer (Uropygi, Arachnida) has been reconstructed from ultra‐thin serial sections. The spatial positions of two trichobothria on the first pair of legs, as well as their innervation by several dendrites point to an effective spatial localization of stimulus sources. Numerous cells are involved in the regeneration of the trichobothrium during moulting. They can be classified according to their position and function in forming the sensillum: there are 11 sensory cells, four inner envelope cells, about 21 trichogen cells and numerous outer envelope cells, including tormogen cells. The complicated cell pattern is compared to conditions in similar sensilla of other arthropods in terms of function and phylogeny.

The infrared receptor of Melanophila acuminata De Geer (Coleoptera: Buprestidae): ultrastructural study of a unique insect thermoreceptor and its possible descent from a hair mechanoreceptor

The paired infrared organs of Melanophila acuminata consist of 50–100 sensilla situated at the bottom of a pit next to the coxae of the mesothoracic legs, where no exocuticle is developed. Each sensillum is accompanied by a wax gland and has a cuticular lens-like spherule (diameter 12–15 μm) bulging out with its upper hemisphere above the surface, covered only by a thin cuticle of about 1 μm. Distal processes of two enveloping cells surround the entire spherule in the form of a flattened protoplasmatic layer with the exception of a small apical stalk connecting the spherule to the outer cuticle. The spherule is innervated by a single sensory neuron of the ciliary type which is anchored ventrally with the distal tip of its cylindrical and unbranched DOS in the spherule. The insertion of the dendrite, which contains a well-developed tubular body, is always eccentric like in a hair mechanoreceptor (sensillum trichodeum) and there is no evidence of any optical function of the spherule. Three enveloping cells exist, but only one — probably the trichogen cell — forms a relatively small outer receptor lymph cavity. In the posterior wall of the pit — where exocuticle is developed — so-called suppressed systems can be found which remain completely below the cuticle with their otherwise well-developed spherules. Additionally, there is a tendency towards basally flattening and longitudinally stretching of spherules which are situated more peripherally. They strongly resemble the basal regions of hair mechanoreceptors (sensilla trichodea) in their immediate neighbourhood which are also accompanied by wax glands. Because of the existence of these transitional stages and the great ultrastructural resemblance between infrared receptors and hair mechanoreceptors concerning the bauplan of the sensory neurons and their mode of innervating the cuticular apparatus, we conclude that the infrared sensilla are probably derived from hair mechanoreceptors. Based on these results and transmission measurements of infrared radiation through the cuticular components of the organ, a model of the possible function of the infrared receptor is presented.

Electron microscope investigation of polytene chromosomes in the Mediterranean fruit fly Ceratitis capitata.

Ultrastructural analyses of polytene chromosomes from male pupal orbital bristle cells and from larval salivary glands of Ceratitis capitata were carried out. It was shown that chromatin complexes corresponding to the X chromosome heterochromatic network are surrounded by material containing ribonucleoprotein (RNP) granules 250-300 A (1 A = 0.1 nm) in diameter. RNP granules of similar size surround the spherical Y chromosome. These data point out the presence of transcriptional activity in both of these chromosomes. The absence of clear structure in chromosomal regions situated between large bands in both types of tissues was observed. These results support the hypothesis of weak synapsis between chromatids or small chromomeres of polytene chromosomes in this species. In addition, we describe a specific puff revealed in both orbital trichogen cells and salivary glands that is morphologically similar to the 93D puff of Drosophila melanogaster.

Antagonistic activities of Suppressor of Hairless and Hairless control alternative cell fates in the Drosophila adult epidermis.

Successive alternative cell fate choices in the imaginal disc epithelium lead to the differentiation of a relatively invariant pattern of multicellular adult sensory organs in Drosophila. We show here that the activity of Suppressor of Hairless is required for both the sensory organ precursor (SOP) versus epidermal cell fate decision, and for the trichogen (shaft) versus tormogen (socket) cell fate choice. Complete loss of Suppressor of Hairless function causes most proneural cluster cells to accumulate high levels of the achaete and Delta proteins and to adopt the SOP fate. Late or partial reduction in Suppressor of Hairless activity leads to the apparent transformation of the tormogen (socket) cell into a second trichogen (shaft) cell, producing a 'double shaft' phenotype. We find that overexpression of Suppressor of Hairless has the opposite phenotypic effects. SOP determination is prevented by an early excess of Suppressor of Hairless activity, while at a later stage, the trichogen (shaft) cell is transformed into a second tormogen (socket) cell, resulting in 'double socket' bristles. We conclude that, for two different cell fate decisions in adult sensory organ development, decreasing or increasing the level of Suppressor of Hairless function confers mutant phenotypes that closely resemble those associated with gain and loss of Hairless activity, respectively. These results, along with the intermediate SOP phenotype observed in Suppressor of Hairless; Hairless double mutant imaginal discs, suggest that the two genes act antagonistically to commit imaginal disc cells stably to alternative fates.

Morphology and ontogeny of single-walled multiporous sensilla of hemimetabolous insects

Comparative morphological investigations were made to determine the common organization plan of single-walled multiporous sensilla. The development of multiporous chemoreceptive sensilla of Gryllus, Oncopeltus and Lepisma follows the same path. Each chemoreceptive sensillum is associated with four types of enveloping cell. During ontogeny, enveloping cell 1 secretes the dendritic sheath. Enveloping cell 4 builds the connection of the hair base with the antennal cuticle. In Gryllus and Oncopehus, enveloping cells 2 and 3 build the hair shaft, the wall pores and pore tubules in nearly equal parts. Enveloping cells 2 and 3 lie side by side in the hair process, in which enveloping cell 2 produces the inner part, enveloping cell 3 the outer part of the hair shaft. In Lepisma the predominant part of the hair shaft with the wall pores is formed by the doubled enveloping cells 3. Interpreting our findings and the literature data, a new proposal is given for the homology of the enveloping cells. In singlewalled chemoreceptors, enveloping cell 1 is considered as thecogen and enveloping cell 4 as tormogen cell. Enveloping cell 2 is interpreted as inner trichogcn cell and enveloping cell 3 as outer trichogen cell.

Polytene chromosomes of monogenic and amphogenic Chrysomya species (Calliphoridae, Diptera): analysis of banding patterns and in situ hybridization with Drosophila sex determining gene sequences.

Standard maps for the five banded polytene chromosomes found in trichogen cell nuclei of the monogenic blowfly Chrysomya rufifacies and the amphogenic Chrysomya pinguis are presented. The chromosomes are highly homologous in the two species; differences in banding patterns are predominantly caused by one pericentric and ten paracentric inversions. In chromosome 5 of the amphogenic Chrysomya phaonis, also analysed in this paper, an additional paracentric inversion was observed. The distribution of species specific inversions indicates that the monogenic C. rufifacies is phylogenetically older than the amphogenic species. The maternal sex realizer locus F'/f on polytene chromosome 5 of C. rufifacies is not associated with a structural heterozygosity. Chromosome pair 6 of C. rufifacies and the sex chromosome pair of C. pinguis are under-replicated in polytene nuclei; they consist of irregular chromatin granules, frequently associated with nucleolus material. Evolution of heteromorphic sex chromosomes in Chrysomya is probably correlated with heterochromatin accumulation. A search for sex determining genes in Chrysomya was initiated using sex determining sequences from Drosophila melanogaster for in situ hybridization. The polytene band 41A1 on chromosome 5 of monogenic and amphogenic Chrysomya species contains sequences homologous to the maternal sex determining gene daughterless (da). Homology to the zygotic gene Sex-lethal (Sxl) of Drosophila is detected in band 39A1 on chromosome 5 of C. rufifacies. The findings reported here are the first evidence for a possible homology between the da gene of Drosophila and the maternal sex realizer F' of C. rufifacies. An hypothesis for the evolution of the maternal effect sex determination of C. rufifacies is proposed.