The structure and function of the bursa copulatrix of the monarch butterfly (Danaus plexippus)

The bursa compulatrix of the Monarch butterfly was investigated utilizing light microscopy, histochemistry, and scanning and transmission electron microscopy in order to relate its morphology to the release of sperm from the spermatophore. The bursa has a row of large chitinous teeth on either side of the organ. The dorsal and ventral surfaces are covered with chitinous plates, the plates having bristles on one side. A single layer of cells lies under both the plates and teeth, one columnar cell under each plate, one cuboidal cell under each tooth. The toothed area has no muscle cells. However, the dorsal and ventral hemispheres of the bursa each have a crescent-shaped packet of muscle fibers that traverse the organ; there are no longitudinal fibers. Spermatophores with thick walls were found in the bursal lumen. Morphological evidence suggests that the presence of the spermatophores is sensed by the bristles and that the packets are opened by contraction of the muscles bringing the large teeth into contact with the spermatophore wall.     

Functional morphology and biomechanics of cuticular fracture at the elytrophoral autotomy plane of the scaleworm Alentia gelatinosa (Annelida: Polynoidae)

Abstract. Autotomy of the elytra (scales) in the annelid Alentia gelatinosa occurs at a breakage plane near the junction between the elytron and its elytrophore (stalk), and requires fracture of the external epidermal cuticle. The mechanism of cuticular fracture was investigated by light and electron microscopy, glycoconjugate histochemistry, direct observation of autotomy in isolated preparations, and mechanical tests. The breakage plane crosses the elytrophoral wall at a cuticular thickening and passes through the subelytral cavity between the elytron and the terminal septum of the elytrophore. At the cuticular breakage zone (CBZ), the collagenous framework of the normal cuticle is replaced with non‐collagenous microfibrils. The CBZ has a complex glycoconjugate composition and includes a strongly sulfated, uronic acid‐containing glycosaminoglycan and a high proportion of disulfide or sulfydryl linkages. Tonofilament‐rich epidermal cells (tendon cells) are attached to the thick cuticle on the dorsal and ventral sides of the CBZ. Dorsal tendon cells have long processes that extend into the elytron near the roof of the subelytral cavity. Ventral tendon cells are linked by connective tissue to the longitudinal and terminal sphincter muscles of the elytrophore. Mechanical tests showed that the elytrophoral wall is not inherently weaker at the autotomy plane than elsewhere. It is hypothesized that at autotomy (i) contractile force generated by the sphincter muscle is transmitted through elytrophoral tendon cells to the ventral side of the CBZ and (ii) contraction of the longitudinal and main circular muscles of the elytrophore increases hydrostatic pressure in its lumen, everts the terminal septum, and generates tension that is transmitted through elytral tendon cells to the dorsal side of the CBZ. This results in stress concentration at the basal edge of the CBZ and initiates fracture. The distinctive microstructure and macromolecular composition of the CBZ may reduce its fracture toughness and make it more susceptible to brittle failure.     

Fine structure of the spermatheca and of the accessory glands in Orchesella villosa (Collembola, Hexapoda)

The spermatheca and the accessory glands of the collembolan Orchesella villosa are described for the first time. Both organs exhibit ultrastructural differences, according to the time of the intermolt in which the specimens were observed. A thick cuticular layer lines the epithelial cells of the accessory glands. In the reproductive phase, they are involved in secretory activity; a moderately dense secretion found in the apical cell region opens into the gland lumen. Cells with an extracellular cistern are intermingled with the secretory cells. These cells could be involved in fluid secretion, with the secretory product opening into the cistern which is filled with an electron-transparent material. After the reproductive phase, the gland lumen becomes filled with a dense secretion. The accessory gland secretion may play a protective role towards the eggs. The spermatheca is located between the accessory glands; its epithelium is lined by a thin cuticle forming spine-like projections into the lumen and consists of cells provided with an extracellular cistern. Secretory cells, similar to those seen in the accessory glands, are missing. Cells with a cistern could be involved in the production of a fluid secretion determining sperm unrolling and sperm motility.     

The Eltringham Organ and a New Thoracic Gland: Ultrastructure and Presumed Pheromone Function (Insecta, Myrmeleontidae)

Elofsson, R. and Löfqvist, J. (Zoological Institute, University of Lund, S-223 62 Lund, Sweden.) The Eltringham organ and a new thoracic gland: ultrastructure and presumed pheromone function . Zool. Scr. 3 (1): 31–40, 1974.—Eltringham's organ is a club-like projection from the posterior margin of hind-wings of some myrmeleontid insects. It bears laterally directed setae, each having a specialized epidermal cell beneath, which gives off secretion into a sub-setal lumen. The lumen continues into the wall of the moat-like socket of the setae. Eltringham's organ fits into an excavation of the lateral body wall connected with the opening of a thoracic gland. The thoracic gland in these animals has not previously been described. It consists of a hollow tube, feebly developed in females. The tube wall houses two cell types: gland cells and cuticular cells. The latter form the cuticle of the general duct lumen and a specialized duct leading from each gland cell. The gland cells have a microvilli-filled cavity which collects the secretion. The thoracic gland produces a pheromone secretion whose distribution is aided by Eltringham's organ where present.     

Structural investigation of the female genitalia and sperm-storage sites in the terrestrial isopod Armadillidium vulgare (Crustacea, Isopoda)

The cuticular genitalia of the terrestrial isopod, Armadillidium vulgare, have two distinct states during the reproductive cycle of the females. The structural differences between the reproductive and non-reproductive states, and the structure of the sperm storage sites were investigated employing electron and light microscopy. In both states the genitalia consist of a distal segment that connects to the gonopore, and a cuticular tube-like structure lining the lumen of the oviduct in the middle region of the oviduct. Sheath-like projections, apparently consisting of cuticular material, extend laterally along two sides of the cuticular tube. In the proximal region of the oviduct cuticular structures are lacking. In the non-reproductive state the distal segment consists of endo-, exo- and epicuticle. The exocuticle is three layered with unusual spongy and dense layers at the distal side. On one side the endocuticle doubles in thickness to form a cuticular bulge that fills the lumen of the distal segment leaving just a narrow U-shaped space. The cuticular tube consists of endo- and epicuticle only. In the reproductive state the distal segment is funnel-shaped and forms branched cuticular folds that increase in complexity from distal to proximal. In the cuticular tube these folds tightly fill the lumen of the oviduct. At the confluence of the oviduct with the ovary spermatozoa are stored in a seminal receptacle.     

Ejaculatory duct of the migratory grasshopper,Melanoplus sanguinipes (fabr.) (Orthoptera : Acrididae): A histological and ultrastructural analysis

The ejaculatory duct of the migratory grasshopper (Melanoplus sanguinipes [Fabr.]) (Orthoptera : Acrididae) is divisible into 3 regions: upper ejaculatory duct (UED) into whose anterior end the accessory glands and vasa deferentia empty; the funnel characterized by its slit-like lumen; and the lower ejaculatory duct (LED). Anteriorly, the UED has a keyhole-shaped lumen surrounded by a thin intima and highly columnar epithelial cells whose most conspicuous feature is massive aggregations of microtubules. More posteriorly, the UED lumen differentiates into dorsal and ventral chambers, the former having a thick cuticular lining armed with spines. In the hindmost part of the UED, the ventral chamber expands to obliterate the dorsal chamber; its cuticular lining thickens, and conspicuous lateral evaginations develop. The thick cuticle includes 3 distinct layers and on its surface carries numerous spatulate processes. In this region, the epithelial cells develop numerous short microvilli beneath which are many mitochondria. As the funnel is reached, the intima becomes extremely thick, and the epithelial cells lack microvilli and most microtubules. Within the funnel, a new, very distinct form of cuticle appears, which is in “units”, each associated with an epithelial cell and having a rounded epicuticular cap. The new cuticle arises ventrally but rapidly spreads to encircle the entire lumen, at which point the LED is considered to begin. Beneath this new cuticle, the epithelial cells are columnar, have long microvilli, numerous mitochondria in the apical cytoplasm, and rough endoplasmic reticulum basally. Apically, adjacent cells are tightly apposed; however, prominent intercellular channels develop more basally. The ejaculatory duct's features are briefly discussed in terms of its role in spermatophore formation.     

Wing-hearts in Mecoptera (insecta)

The accessory pulsatile organs for hemolymph circulation in the wings of 7 Mecoptera species were investigated by means of serial semi-thin sections, SEM and TEM. The wing-hearts are located in the dorsal meso- and metathorax, and have no connection to the aorta. Each wing-heart consists of a small hemolymph chamber formed above by the convex scutellum, and below by a horizontal muscular diaphragm. The chamber is connected to the posterior wing veins by a cuticular tube on each side of the body. The diaphragm (10–15 μm thick) is convex in cross-section and consists of transversely extended muscle fibers. Their ultrastructure reveals typical characters of myocardial and other visceral muscle fibers. The diaphragm muscle is innervated by a pair of thin nerves originating from the thoracic ganglion of each corresponding segment. The diaphragm is held in a convex position by numerous elastic strands (2 μm in diameter), which extend through the wing-heart lumen between the scutellum and the diaphragm. The diastolic phase of the wing-heart is caused by contraction of the diaphragm muscle fibers. Thus, the diaphragm flattens and hemolymph is drawn from the posterior wing veins. The systolic phase is caused by the elasticity of the suspending strands after relaxation of the muscle fibers. The elastic strands pull the diaphragm back into convex position and hemolymph is expelled out of the scutellum lumen into the thorax cavity through a valvular opening on the anterior side. The hemolymph flow from the posterior wing base to the scutellum lumen, was visualized by staining the hemolymph. In Panorpa communis the volume of the wing-heart lumen measures 1.6 × 10⁻² mm³ in the mesothorax, and 1.2 × 10⁻² mm³ in the metathorax. Each heartbeat transports a maximum of 65% of these volumes. The pumping frequency was 78 ± 20 beats per min, registered with a non-invasive photo-optical method in restrained animals. Corresponding pulsating movements occur as a passive phenomenon of wing-heart activity in a distinct area of the wing base. Only minor differences were found in the construction of wing-hearts among the investigated species, except for Boreus hyemalis, which lacks these accessory circulatory organs. The functional morphology of the wing-hearts in Mecoptera is compared with that of other Holometabola and aspects of the evolution of these organs are discussed.     

The ultrastructure of the female accessory gland,the cement gland,in the insect Rhodnius prolixus

The cement gland of Rhodnius prolixus is an epidermally derived tubular gland consisting of a distal synthetic region and a proximal muscular duct region. The synthetic region consists of numerous secretory units joined to a central chitinous duct via cuticular ductules. Proteinaceous secretion, synthesized by the goblet-shaped secretory cell, passes through the delicate cuticular lattice of a ductule-end apparatus and out through fine ductules to the central duct. Secretory cells are rich in rough endoplasmic reticulum and mitochondria. Light microscopy, SEM and TEM reveal the delicate lattice-like end apparatus structure, its formation and relationship to the secretory cell. The secretory cell associates via septate junctions with a tubular ductule cell that encloses a cuticle-lined ductule by forming an elaborate septate junction with itself. The ductules are continuous with the cuticle lining of the large central duct that conveys secretion to the proximal area. The proximal muscular duct has a corrugated cuticular lining, a thin epithelium rich in microtubules and thick longitudinal, striated muscles which contract during oviposition, forcing the secretion out. Histochemistry and electrophoresis reveal the secretion as proteinaceous.     

Fine structure of the prothoracic mycangium, a chamber for the culture of symbiotic fungi, in the southern pine beetle, Dendroctonus frontalis

The ultrastructure of the prothoracic mycangium of female Dendroctonus frontalis is examined. The mycangium consists of a cuticular invagination within which symbiotic fungi are cultured by the pine beetle and transported to new host trees. Secretions from two types of gland cells pass into the mycangial lumen. The plasma membrane of type-1 cells is invaginated to form an enclosed extracellular cavity. The secretory product passes into the cavity, then through fine cylindrical channels into an end apparatus and finally via an efferent cuticular ductule to the lumen of the mycangium. Secretion of the type-2 cells is released into a cavity just beneath the mycangial cuticle. The cuticle over this cavity is quite thin (1-2mu), consisting mostly of inner epicuticle riddled with irregular canals through which the secretion reaches the lumen. Beneath the patches of porous cuticle are ribs (up to 1Omu in thickness) which flank the cavities and presumably provide structural support for the porous secretory zones.     

The fine structure of the somatic muscles and their attachment to the cuticle in two species of Pycnogonida

The fine structure of the somatic muscles and their attachment to the cuticle in the pyenogonids Nymphon (Chaetonymphon) macronyx G. O. Sars and Boreonymphon cf. abyssorum (Norman) is described. The muscles possess characteristics which are typical of arthropod slow muscle fibers: relatively long sarcomeres, a mean A-band length of about 6 μm and a ratio of thin to thick contractile filaments of 4:1. The sarcotubular system consists of distinct t-tubules, an irregular SR part and randomly distributed dyads and triads, the muscles are attached to the cuticle by specialized epidermal cells containing microtubules extending from the cuticular to the muscular side. The myoepidermal and epidermal-cuticular junctions are described.     

Fine structure of the tip of the labellar taste hair of the blow flies,Phormia regina (Mg.) and Calliphora vicina R.-D. (Diptera,Calliphoridae)

Summary The labellar taste hairs of the blow flies, Phormia regina and Calliphora vicina, have an opening mechanism at the tip which consists of two stump cuticular prongs and a funnel-like cuticular pouch. Opening and folding of these structures are regulated by the pressure within the dendrite-free lumen of the hair. The extrusion of viscous substance at the tip of the taste hair is possible through spongy cuticle and one pore in each prong; it seems likewise to depend on the pressure within the dendrite-free lumen and results in regional collapsing of this lumen. Described and discussed are: The cuticle and pores of the structures at the hair tip, pore filaments which extend from the dendrites, and the number and arrangement of the dendrites.This work was supported by a grant from the 7USDA, Entomology Research Division, Beltsville, Md., and the grant GB-13500 from the National Science Foundation.We thank Dr. J. F. Worley, USDA, Plant Science Research Division, Beltsville, Md., for his collaboration in fluorescence microscopy.     

Formation of the hindgut cuticular lining during embryonic development of Porcellio scaber (Crustacea,Isopoda)

The hindgut and foregut in terrestrial isopod crustaceans are ectodermal parts of the digestive system and are lined by cuticle, an apical extracellular matrix secreted by epithelial cells. Morphogenesis of the digestive system was reported in previous studies, but differentiation of the gut cuticle was not followed in detail. This study is focused on ultrastructural analyses of hindgut apical matrices and cuticle in selected intramarsupial developmental stages of the terrestrial isopod Porcellio scaber in comparison to adult animals to obtain data on the hindgut cuticular lining differentiation. Our results show that in late embryos of stages 16 and 18 the apical matrix in the hindgut consists of loose material overlaid by a thin intensely ruffled electron dense lamina facing the lumen. The ultrastructural resemblance to the embryonic epidermal matrices described in several arthropods suggests a common principle in chitinous matrix differentiation. The hindgut matrix in the prehatching embryo of stage 19 shows characteristics of the hindgut cuticle, specifically alignment to the apical epithelial surface and a prominent electron dense layer of epicuticle. In the preceding embryonic stage – stage 18 – an electron dense lamina, closely apposed to the apical cell membrane, is evident and is considered as the first epicuticle formation. In marsupial mancae the advanced features of the hindgut cuticle and epithelium are evident: a more prominent epicuticular layer, formation of cuticular spines and an extensive apical labyrinth. In comparison to the hindgut cuticle of adults, the hindgut cuticle of marsupial manca and in particular the electron dense epicuticular layer are much thinner and the difference between cuticle architecture in the anterior chamber and in the papillate region is not yet distinguishable. Differences from the hindgut cuticle in adults imply not fully developed structure and function of the hindgut cuticle in marsupial manca, possibly related also to different environments, as mancae develop in marsupial fluid. Bacteria, evenly distributed within the homogenous electron dense material in the hindgut lumen, were observed only in one specimen of early marsupial manca. The morphological features of gut cuticle renewal are evident in the late marsupial mancae, and are similar to those observed in the exoskeleton.     

Drosophila Knickkopf and Retroactive are needed for epithelial tube growth and cuticle differentiation through their specific requirement for chitin filament organization

Precise epithelial tube diameters rely on coordinated cell shape changes and apical membrane enlargement during tube growth. Uniform tube expansion in the developing Drosophila trachea requires the assembly of a transient intraluminal chitin matrix, where chitin forms a broad cable that expands in accordance with lumen diameter growth. Like the chitinous procuticle, the tracheal luminal chitin cable displays a filamentous structure that presumably is important for matrix function. Here, we show that knickkopf (knk) and retroactive (rtv) are two new tube expansion mutants that fail to form filamentous chitin structures, both in the tracheal and cuticular chitin matrices. Mutations in knk and rtv are known to disrupt the embryonic cuticle, and our combined genetic analysis and chemical chitin inhibition experiments support the argument that Knk and Rtv specifically assist in chitin function. We show that Knk is an apical GPI-linked protein that acts at the plasma membrane. Subcellular mislocalization of Knk in previously identified tube expansion mutants that disrupt septate junction (SJ) proteins, further suggest that SJs promote chitinous matrix organization and uniform tube expansion by supporting polarized epithelial protein localization. We propose a model in which Knk and the predicted chitin-binding protein Rtv form membrane complexes essential for epithelial tubulogenesis and cuticle formation through their specific role in directing chitin filament assembly.     

Studies on the tardigrades. II. Fine structure of the pharynx of milnesium tardigradum doyère

The fine structure of the pharynx and associated valve of the tardigrade Milnesium tardigradum is presented. The pharynx consists of a triradiate, cuticle lined lumen surrounded by radially arranged muscle cells and special apical cells which cap each of the ventricles of the lumen. The valve is an unusual structure marking the anterior limit of the pharynx. It is a specialization of the cuticular lining of the foregut and the apical cells of the pharynx. The significance and phyletic affinities of these structures are discussed.     

Specialization of the hindgut wall for the attachment of symbiotic micro-organisms in a termite Procubitermes aburiensis (Isoptera,Termitidae, Termitinae)

The posterior colon of worker-caste termites accommdates an abundant, heterogeneous population of procaryotic organisms which are retained by attachment to prominent cuticular spines elaborated from the gut wall. The spines extend to nearly one half the diameter of the lumen and are each supported by a specialized root cell in which bundles of parallel microtubules traverse the cytoplasm from the apical to the basal surfaces. Additional epithelial cells are present which show infoldings of the apical plasma membrane and are overlain by cuticle containing deep, vase-shaped pits opening to the gut lumen. It is proposed that the root cells are designed to resist shearing forces transmitted to the base of each spine during contractions of the gut. The cuticular pits may represent sites of permeability to the end products of microbial metabolism.     

Histology of adult Litomosoides carinii (Nematoda: Filarioidea)

The histology of male and female Litomosoides carinii was studied in serial sections. The body wall of the female worm consists of a rather thin cuticle, very extended lateral hypodermal chords and small sectors of shallow coelomyarian muscle cells. In the female worm an intestinal lumen is found, which is filled with material throughout the worm's length. Both ovaries are situated close to the posterior end. In both genital tubes all stages of oogenesis and embryogenesis could be observed in approximately equal stages of development in most of the sections. The lateral chords of the male worm are very small, whereas the somatic muscle cells are large and of the circomyarian type. The intestine is a minute tube beside the much thicker testis and vas deferens. Considerable sexual dimorphism could be observed in almost all organs of adult Litomosoides carinii.     

A new type of brachyuran seminal receptacle in the masked crab Ethusa mascarone (Brachyura,Ethusidae)

The reproductive system of the female Ethusa mascarone was studied with a combination of histological and MRI‐techniques. The study reveals a completely new type of eubrachyuran seminal receptacle. This receptacle consists of two largely separate chambers that engage with each other in a manner similar to shaking hands. One chamber facing the medial axis is lined by cuticle while the second chamber consists of a thick holocrine epithelium. Both chambers are connected by two openings of a unique structure. First, the glandular chamber opens ventro‐laterally to the cuticle chamber via a laterally flattened connective duct that is lined by a highly folded cuticle. A second opening connects both chambers dorsally with the oviduct orifice. A distinct character is the cuticular hook‐like projection that is situated in between the connection of oviduct opening, the glandular chamber and the cuticle chamber of the seminal receptacle. The complete seminal receptacle exhibits a combination of plesiomorphic and apomorphic characters. The arrangement of the receptacles featured in two separate chambers, including the ventro‐lateral connection of the glandular chamber to the cuticle chamber, presumably reflects an early evolutionary stage of an eubrachyuran receptacle. In contrast, the dorso‐lateral opening between both chambers, including the hook‐like projection, appears to be an apomorphic character of at least E. mascarone. J. Morphol. 277:1497–1508, 2016. © 2016 Wiley Periodicals, Inc.     

Tarsal attachment devices of the southern green stink bug Nezara viridula (Heteroptera: Pentatomidae)

Based on analyses with cryo‐scanning and transmission electron microscopy, the present study reports on the morphology and ultrastructure of the attachment structures of the green stinkbug Nezara viridula L. (Heteroptera: Pentatomidae), a cosmopolitan pest of different crops in most areas of the world. In addition, the presence and distribution of large proportions of the elastic protein resilin in these structures was revealed by confocal laser scanning microscopy. The attachment structures of each leg comprise two sclerotised claws, a pair of smooth flexible pulvilli and a hairy adhesive pad located at the ventral side of the basitarsus. No sexual dimorphism is evident. Contact areas of resting individuals on a smooth surface show that N. viridula creates contact to the substrate with the ventral surface of (a) the distal portions of the pulvilli, (b) the setae of the hairy adhesive pad, (c) the two paraempodia representing mechanosensory setae, and (d) the tips of the claws. Each pulvillus is a sac‐like structure formed by complex cuticular layers that vary in their structure and resilin content. The dorsal side consists of sclerotised chitinous material, while the ventral cuticle consists mainly of resilin and shows a very thin epicuticle and a thick exocuticle. The setae of the hairy adhesive pad are pointed and socketed. They exhibit a pronounced longitudinal gradient in the material composition, with large proportions of resilin being present in the setal tips. In most of these setae, especially in those of the distal‐most part of the pad, also a transverse gradient in the material composition is visible.     

Ultrastructure of the accessory gland in the parthenogenetic thrips heliothrips haemorrhoidalis (Bouché) (Thysanoptera : Thripidae)

The fine structure of the reproductive accessory gland of the parthenogenetic thrips Heliothrips haemorrhoidalis (Thysanoptera : Thripidae) is reported. It consists of an apical bulb and a fine gland duct. The former consists of an epithelium with secretory and duct-forming cells surrounding a large gland lumen lined with a thin cuticle and filled with dense secretion. Spent secretory cells degenerate and are eliminated from the epithelium. The gland duct is characterized by an irregular, branched lumen surrounded by a very flat epithelium. A valve controls the opening of the duct lumen. The proximal gland duct runs through a cuticular papilla that opens between the dorsal ovipositor valves. The secretions may serve for ovipositor valve lubrication and possibly to protect laid eggs. Observations of serial sections through the vagina exclude the presence of a spermatheca in this species.