The structure of the antennae of the Florida Queen butterfly, Danaus gilippus berenice (Cramer)

Interest in the structure of the antennae of the Florida Queen butterfly arises from the finding that a pheromone is active in their courtship. Light and electron microscopic techniques were used to study the sensilla on the antennae and three types of sensilla with perforated walls were identified. The most common of these are short, thin-walled pegs which are distributed over most of the antennal surface. Long, curved, thin-walled pegs occur in patches on the inner medial antennal surface. Multiple coeloconic sensilla are present having up to 50 pegs in one sensillum. On the outer 28 flagellar subsegments there are two such sensilla per subsegment. In addition there are on the antennae long, thick-walled hairs which are mechanoreceptors and probably also contact chemoreceptors. Sunken pegs, the function of which is not known, occur on the antennae. Grooved sensilla were found with the electron microscope but could not be identified with the light microscope. There was no indication of sexual dimorphism in sensilla types or numbers on the antennae.     

The structure of the rectal pads of Periplaneta americana L. With regard to fluid transport

The rectum of Periplaneta americana L. is lined with cuticle and has six radially arranged cushion-shaped thickenings, the rectal pads, composed of columnar cells. Narrow strips of simple rectal cells lie between the pads. Tall junctional cells form a thin but continuous collar around the pads where they join the rectal cells. The epithelium is surrounded by a layer composed of circular and longitudinal muscles and connective tissue. This layer of muscle and connective tissue is innervated and tracheated, and is separated from the pad surface by a subepithelial sinus. Fluid flowing through the sinus enters the haemolymph through openings in the muscle layer whre large tracheae penetrate. These openings can be sealed by muscle contractions that appress the muscle around the openings against the pad surface. The tracheae pass on into the pads, following basement membrne-lined indentations of the pad surface. Within the pad tracheolar cells send fine branches between the cells. Near the apical and basal surfaces the lateral membranes of pad cells are bridged by septate desmosomes that form a continuous band around the cells. Between apical and basal septate desmosomes is an interconnected labyrinthine system of intercellular spaces. There are three kinds of space, dilations and apical sinuses, both of variable size, and narrow communicating channels about 200 Å wide. The membranes of the latter have mitochondria closely associated with them. Continuity between the system of spaces and the subepithelial sinus is established by the basement membrane-lined invaginations of the basal surface where tracheae penetrate between pad cells. Apical surfaces of the pad cells are highly infolded and are also associated with mitochondria. However, unlike the lateral membranes facing the narrow channels, the apical membranes have a cytoplasmic coating of particles. Both associations of mitochondria with membranes constitute discrete structural entities that are found in many transporting epithelia, and we have termed them “plasmalemma-mitochondrial complexes.” As the rectal pads are organized into systems of spaces that ultimately open in the direction of fluid movement, existing models of solute-coupled water transport can be applied. However, the rectal pads are structurally more complex than fluid-transporting tissues of vertebrates. This complexity may be related to the ability of the rectum to withdraw water from ion-free solutions in the lumen. We present a structural model involving solute recycling to explain the physiological characteristics of rectal reabsorption.     

Analysis of mouthpart movements during feeding of Frankliniella occidentalis (pergande) and F. schultzei trybom (Thysanoptera : Thripidae)

Scanning electron microscopy was used to elucidate the morphology and sequential movement of thrips feeding structures in the 2 species, Frankliniella occidentalis and F. schultzei, (Thysanoptera : Thripidae). The mouthcone consists of paired paraglossae, and fringed labral pad, a single, apically fused mandible and a pair of interlocking maxillae. The maxillae are open apically and form a feeding tube. Ten pairs of sensory pegs of 3 distinct morphological types (sensilla basiconica with a cuticular collar, sensilla basiconica without a cuticular collar, and sensilla trichoidea) were found on the paraglossae. The possible function of these sensory structures in host finding and choice are discussed. No structure for rasping the leaf's surface was found on the mouthcone or the ventral surface of the insect. Live specimens were observed feeding through Parafilm on artificial media (sucrose solution) and lettuce leaf tissue. These observations support earlier findings that thrips feed by piercing leaf cells with the mandible and ingesting cell contents through the feeding tube formed by the maxillary stylets. Based on these findings, we suggest that thrips be classified as piercing-sucking rather than rasping-sucking insects.     

TRANSFER CELLS IN THE PLACENTAL PAD AND CARYOPSIS COAT OF PAPPOPHORUM SUBBULBOSUM ARECH. (POACEAE)

During caryopsis development the layers of the pericarp, integuments, and nucellus all contribute to the formation of the caryopsis coat. The coat consists of a layer of outer pericarp epidermal transfer cells, a collapsed and senescent layer of middle pericarp cells, and a discontinuous layer of inner pericarp epidermal transfer cells. The latter is not present across the placental pad. The integuments are present as a collapsed dense layer, the nucellus is discontinuous and cellular. The placental pad occurs at the ventral surface of the caryopsis, opposite the scutellum and coleorhiza. It consists of 15–20 collapsed cell layers, including the pigment strand and placental vascular bundle. From the inside several partially collapsed cell layers of the nucellar projection occur which contain transfer-cell walls. The middle dense layers, the pigment strand, consist of the middle pericarp remnant, plus the remains of the placental vascular bundle. The pericarp inner epidermis does not extend across the pad. The aleurone layer is a continuous uniseriate layer around the entire caryopsis except at the placental pad; here it is crushed and contains the remnant of a transfer-cell wall. The outer pericarp epidermis is a continuous layer of transfer cells across the pad. These cells contain membranous inclusions suggesting that they may be functional during germination.     

The digital pads of the tree frog, Hyla cinerea. II. The mucous glands

The mucous glands consist of a single row of cells surrounded by smooth muscle. The cells are attached at their apical and basal regions and only cytoplasmic projections loosely link the lateral aspects of adjacent cells. Material accumulates in the cisternae of the rough endoplasmic reticulum, appears to form into dense granules in the Golgi apparatus, and then before being secreted, undergoes chemical and morphological alterations. Since some glands secrete onto the dorsal epidermis of the digits, the mucous is believed to function as a wetting agent for the skin as well as an aid to climbing.     

Ultrastructure of the galeal sensory complex in adults of the Colorado potato beetle, Leptinotarsa decemlineata

ABSTRACT. The structure of galeal sensilla of the Colorado potato beetle, Leptinotarsa decemlineata Say (Coleoptera: Chrysomelidae), is described using electron microscopical methods. Previous electro-physiological studies indicate that these sensilla respond to amino acids, sucrose and plant saps. One physiological type is particularly sensitive to L-alanine and gamma amino butyric acid (GABA).
Three morphologically different types of sensilla occur on the galeal tip. The more numerous apical pegs are not distinguishable from one another on the basis of external structure, although they differ physiologically. Five sensory cells are associated with most apical pegs. One apical peg, the α-sensillum, contains only four cells. All apical pegs have one cell with a tubular body. The remaining cells have unbranched dendrites and are associated with a single apical pore.
Apical hairs differ from the apical pegs by having double innervation. Within the hair shaft, a dendritic sheath is lacking and the sensillar sinus extends to the base of the hair. The function of this hair type is not known.
Numerous mechanosensory hairs which surround the other sensilla are singly innervated and contain a tubular body at the level of the outer dendritic segments.     

Structural features of mycorrhizal associations in two members of the Monotropoideae, Monotropa uniflora and Pterospora andromedea

Species in the subfamily Monotropoideae (family Ericaceae) are achlorophyllous and myco-heterotrophic. They have become highly specialized in that each plant species is associated with a limited number of fungal species which in turn are linked to autotrophic plants. This study provides an updated and comprehensive examination of the anatomical features of two species that have recently received attention with respect to their host-fungal specificity. Root systems of Monotropa uniflora and Pterospora andromedea collected from the field were characterized by light microscopy and scanning electron microscopy. All roots of both species were associated with fungi, each root having a well-developed mantle, paraepidermal Hartig net, and intracellular fungal pegs within epidermal cells. The mantle of M. uniflora was multi-layered and numerous outer mantle hyphae developed into cystidia of two distinct morphologies. Large calcium oxalate crystals were present, primarily on the mantle surface. The outer mantle of P. andromedea was more loosely organized, lacked cystidia, and had smaller plate-like as well as cylindrical crystals on the surface and between outer mantle hyphae. Fungal pegs in M. uniflora originated from inner mantle hyphae that penetrated the outer tangential wall of epidermal cells; in P. andromedea, these structures were initiated either from inner mantle hyphae or Hartig net hyphae and penetrated radial walls of epidermal cells. With respect to function, fungal pegs occurred frequently in both host species and, although presumed to be the sites of active nutrient exchange, no direct evidence exists to support this. Differences between these two monotropoid hosts, resulting from the mycorrhizal fungi with which each associates, are discussed.     

Hyphal anastomosis inPyricularia oryzae cav.

Summary Hyphal anastomosis inPyricularia oryzae occurs naturally in the lower epidermal cells and in the vascular bundles of young lesions on rice. In those cells the invading blast fungus are active. Two hyphal cells lying side by side start an anastomosis by forming two, one from each, very short penetration pegs which are opposite to each other. The cell wall of the pegs and the wall in the vicinity of them are then gradually eliminated and thus form a fusion aperture of 0.2–0.6 m in diameter that is big enough for the migration or exchange of nucleus and cytoplasm between two hyphal cells. The explanation of genetical variation inP. oryzae may be sought on the basis of the ultrastructural evidence of hyphal anastomoses presented in this paper.     

Cytological studies on rust fungi. (vi) fine structures of infection process of Kuehneola japonica (diet.) dietel

The behavior of rust fungi in their host plants has been elucidated by electron microscopy. However, most of the ultrastructural studies on rust fungi have focused on the uredial stage. In order to elucidate the features of the sporidial stage, we studied the fine structure of Kuehneola japonica, a short-cycle rust, in rose leaves. Infection pegs arising from appressoria penetrated the host walls. Papillae formed at the time of penetration against the outer epidermal cell walls. The papillae which had formed at the penetration sites grew extensively and partially surrounded the intracellular hyphae which were connected with the infection pegs. The intracellular hyphae in the epidermal cells developed further and entered adjacent parenchyma cells. Walls of parenchyma cells either invaginated or thin papillae formed at penetration sites and the invaginated walls or papillae surrounded the necks of the intracellular hyphae. Intracellular hyphae in both epidermal and parenchyma cells were not enveloped by the sheath before 20 days after inoculation. In specimens prepared 20 days after inoculation, some of the intracellular hyphae were enveloped by a sheath in both palisade and spongy parenchyma cells. The sheathed hyphae resembled haustoria of other rust fungi which had been described previously. Teliospore initials were formed in mycelial masses in intercellular spaces between the epidermal cells and palisade parenchyma cells 20 days after inoculation. Uninucleate teliospores developed from teliospore initials 30 days after inoculation.Contribution No. 32.     

Odour sensitivity of antennal olfactory cells underlying grooved pegs of Anopheles gambiae s.s. and An. quadriannulatus

In female mosquitoes of the anthropophilic species Anopheles gambiae Giles s.s. and the zoophilic An. quadriannulatus Theobald single sensillum recordings from grooved pegs were made. In both species, the majority of these sensilla responded to ammonium hydroxide, butylamine and propanoic acid, whereas a smaller part responded to acetone. Lactic acid, butanone, 3-methyl phenol and 1-octen-3-ol evoked responses in a minority of grooved pegs only. In An. gambiae these four substances evoked either excitatory or inhibitory responses. In An. quadriannulatus excitatory and inhibitory responses were only found on stimulation with lactic acid; butanone, 3-methyl phenol and 1-octen-3-ol only evoked inhibition in the pegs of this species. More than half of the grooved pegs responded to water vapour with an increase in spike frequency. As opposed to this, in some pegs inhibitory responses were found upon stimulation with vapour of low humidity. This suggests that grooved pegs may play a role in humidity perception in Anopheles. Dose-response relations were investigated for cells excited by ammonium hydroxide, butylamine and propanoic acid. Excitatory responses to these three substances were dose-dependent. No significant differences were found between the dose-response curves of the two species. It is concluded that in both species the host odours tested are not perceived by specialist cells. Combined information from generalist cells may provide a detailed `odour profile' of the host.     

Histological changes in the epidermis of the subdigital lamellae of Anolis carolinensis during the shedding cycle

The characteristic anoline climbing organ consists of a number of lamellar scales, on whose outer scale surface are numerous keratinized setae which contact the substrate. These setae are derived from the Oberhautchen of the epidermal generation, and as such are renewed and shed periodically along with the rest of the epidermal material. The histological development of the setae is described, and modifications of the surrounding elements are noted. The relative lengths of the setae and their congregation to form a pad unit poses certain mechanical problems during morphogenesis, simply in terms of accommodation between the functional outer epidermal generation and dermal core of each lamella. Regression of the dermal core and a distal migration of some cells permits accommodation within the lamella for the distal aspect of the Oberhautchen layer, or free margin. Additionally, changes in the gross shape of the lamella occur throughout the sloughing cycle, and a swelling of the cells of the lacunar tissue results in a gap between the stratum corneum of inner and outer epidermal generations. There is a considerable amount of variation in mitotic activity between the germinal layers of opposite sides of the lamella.     

The formation of the cyst wall of the metacercaria of Fasciola hepatica L.

Summary The several concentric layers of the cyst wall of Fasciola hepatica are formed from precursors synthesised in the cystogenic cells of the cercaria during its development in the redia. A cinematographic analysis shows that the separate components are released in succession during encystment.The outer portion of the wall consists of two layers: a tanned protein and a carbohydrate-protein complex. The granular precursors of these are formed in separate groups of cells and migrate from these cells into the superficial epithelium (embryonic epithelium) during development. They are released to form the outer wall by the bursting of the embryonic epithelium at the beginning of encystment. This process is rapid and is completed in a few minutes.A pause follows the separation of the outer wall during which a further polysaccharide layer is released and the cells, which contain the rod-like scrolls of sheets of the laminated component of the inner wall, migrate from within the cercaria through gaps in the superficial musculature on to the cercarial surface to form a new epithelium replacing that previously shed.The cercaria now begins a series of complex oscillatory movements within the enveloping outer cyst wall during which the scrolls are secreted into the space underneath the outer wall, unroll and are compacted by the animal's movements into the lamellar inner wall.The rodlets are enclosed in vacuoles and their secretion is effected by the fusion of the vacuolar membrane with the plasma membrane without destroying the integrity of the cells, which remain to constitute the epithelium of the juvenile fluke when this emerges later.     

The basement-membrane-like matrix of the mouse EHS tumor: I. Ultrastructure

The fine structure of the extracellular matrix was examined in the Engelbreth-Holm-Swarm (EHS) tumor of the mouse. The matrix is composed of layers parallel to the surface of the associated cells; the layers are poorly defined close to the cells (proximal region) but quite distinct at a distance from the cells (distal region). In the proximal region of the matrix, the indistinct layers are composed of three types of structures: 1) a network of 3- to 8-nm-thick "cords" makes up the bulk of the tissue. 2) Within the network are scattered few 7- to 10-nm-wide, hollow rods of indefinite length, referred to as "basotubules"; their cross section has a dense, more-or-less-circular or -pentagonal wall and a light lumen containing a spherule. In addition, many pale profiles similar to these cross sections are present; they are interpreted as small, independent structures. 3) Minute structures composed of two parallel, 3.5-nm rodlets are referred to as "double pegs." In the distal region of the matrix, the distinct layers include the same three types of structures, but basotubules are numerous and prominent; in each layer, they are arranged in picket-fence fashion along two parallel planes. Cords are packed between them. Double pegs are scattered throughout the clear interlayer spaces. Inasmuch as cord network, basotubules, and double pegs are present in the two regions of the tumor matrix, both regions resemble basement membrane. To explain the contrast between the paucity of basotubules in the proximal region and their abundance in the distal region, it is proposed that, as the production of newer matrix by the cells causes the older matrix to be displaced distally, the independent structures seen as pale profiles in the proximal region gradually assemble into basotubules.     

Heat-induced blebbing and vesiculation of the outer membrane of Escherichia coli.

Thermal damage to the outer membrane of Escherichia coli W3110 was studied. When E. coli cells were heated at 55 degrees C in 50 mM Tris-hydrochloride buffer at pH 8.0, surface blebs were formed on the cell envelope, mainly at the septa of dividing cells. Membrane lipids were released from the cells during the heating period, and part of the released lipids formed vesicle-like structures from the membrane. This vesicle fraction had a lipopolysaccharide to phospholipid ratio similar to that of the outer membrane of intact cells, whereas it had a lower content of protein than the isolated outer membrane. After heating bacterial cells at 55 degrees C for 30 min, the resulting leakage from the cells of a periplasmic enzyme, alkaline phosphatase, amounted to 52% of the total activity, whereas no release of a cytoplasmic enzyme, glucose-6-phosphate dehydrogenase, was detected. The results obtained suggest that surface blebs formed by heat treatment almost completely consist of the outer membrane and that the blebs may be gradually released from the cell surface into the heating menstruum to partially form vesicles.     

The bovine tubouterine junction: general organization and surface morphology

The bovine tubouterine junction is composed of three parts (terminal tubal segment, transition region proper, uterine apex) and follows a sigmoidal course displaying a tubal and an uterine curvature. In the terminal tubal segment, 4–8 primary longitudinal folds and a system of lower secondary folds, ridges and chords project into the centrally located lumen. The transition region proper possesses a slit-like lumen because of the existence of a thick mucosal pad containing the first uterine glands. The longitudinal primary folds of the tube broaden, flatten and start to diverge when they reach the transition region proper. The mucosal pad and broadened folds are heavily vascularized. A system of lateral outpocketings with blind ends pointing in an ampullary direction develops between the primary and secondary folds, the ridges and chords of the terminal tubal segment and transition region proper. From the bottom of these outpocketings, short tubulo-alveolar crypts originate. The mucosa of the uterine apex forms low transversal ridges. The musculature of the bovine tubouterine junction is divided into a continuous circular or spiral intermediate layer, flanked by inner and outer longitudinal layers. The outer longitudinal layer is incomplete in the terminal tubal segment but increases in thickness to form a continuous stratum in the uterine apex. An inner longitudinal layer occurs only in the terminal tubal segment where it is best developed in the bases of the primary longitudinal folds. The simple columnar surface epithelium of the tubouterine junction contains ciliated and non-ciliated cells. The former undergo cyclical changes, and increase during estrus and postestrus. During proestrus, groups of non-ciliated cells display bulbous apical protrusions. During proestrus and estrus, circumscribed epithelial lesions expose the underlying basal lamina.     

Structural organization of the toe pads in the amphibian Philautus annandalii (Boulenger, 1906)

We describe the morphology of toe pads in the Himalayan tree frog Philautus annandalii. These are expanded tips of digits and show modifications of their ventral epidermis for adhesion. The outer cells of toe pad epidermis (TPE) bear surface microstructures (0.7 × 0.2 μm), which are keratinized. Their cytoplasm contains no organelles, but pleomorphic nuclei and mucous granules (0.4–0.5 μm) that glue the keratin filaments. In the intermediate cell layer of TPE, similar keratinized microstructures as in the outer cells are present, so that when the outer layer is shed, it is ready with features for adhesion. These cells contain more keratin than the outer cells. The basal cell layer contains thin keratin bundles and usual cell organelles. The dermis contains mucous‐secreting glands, whose ducts open in the outer epidermal cell layer in channels. The dorsal epidermal cells lack surface microstructures and keratin bundles. Ultrastructural features suggest that toe pads utilize the surface microstructures for adhesion aided by mucus, in which the intermediate cell layer seems to bear the shear stress generated during locomotion. Further, TPE can expand and fit into an increased contact area of the substrate. The long, surface microstructures may also help in mechanical interlocking with rough surfaces on plants.     

Extracellular metabolism of cyclic AMP.

Intravenously administered cyclic [8-3H]AMP to rats was quickly eliminated from the circulation. After 2 min 93% of the administered radioactivity disappeared from the plasues was recovered mainly in the form of nucleotides, ATP, ADP, AMP and IMP. In vitro contact of cyclic AMP with perfused liver, isolated liver cells and adipose tissue resulted in a rapid breakdown of the nucleotide, presumably on the outer surface of the cells. The degradation products have been identified mainly as adenosine and inosine. Incubation of adipose tissue and isolated liver cells with [3H] AMP also resulted in the breakdown of the nucleotide in themedium. The rate of AMP degradation by these tissues was faster than that for cyclic AMP degradation. The data suggest that cyclic AMP is readily metabolized on the outer surface of cells to products which may be converted within the cells to nucleotides. These findings seem of importance for the quantitative assessments of cellular cyclic AMP outflow during hormonal stimulation.     

Intercellular adhesion in the cellular slime mold Polysphondylium pallidum inhibited by interaction of asialofetuin or specific univalent antibody with endogenous cell surface lectin.

Previous work has shown that, as amoebae of the cellular slime mold Polysphondylium pallidum become aggregation competent, they accumulate on their cell surface a carbohydrate-binding protein (lectin) named pallidin. These amoebae also possess cell surface receptors, presumed to contain complex oligosaccharides with a high affinity for the endogenous lectin. If lectin-receptor interactions mediate cell-cell contact, then appropriate concentrations of pallidin inhibitors should block cell cohesion. Two potent macromolecular antagonists of the lectin were employed: the desialylated form of the glycoprotein fetuin and the univalent antibody (Fab) prepared against pallidin. We studied the effects of these inhibitors on rotation-mediated aggregation of P. pallidum amoebae under a variety of assay conditions. Amoebae exposed to hypertonic conditions or to antimetabolites (“Permissive conditions”) were selectively blocked from associating by microgram quantities of the lectin inhibitors, whereas cells in isotonic buffer (“nonpermissive condition”) were only slightly affected. A comparison of the morphology of agglutinates formed under the various conditions allows several explanations for the different susceptibilities to inhibition by antipallidin reagents. Although not conclusive, the work supports a model of cell adhesion in this simple eukaryotic system based at least in part on specific interactions between carbohydrate-binding proteins and receptors on adjoining cells.     

Functional morphology of a double-walled multiporous olfactory sensillum: the sensillum coeloconicum of Bombyx mori (Insecta, Lepidoptera)

The fine structure of coeloconic sensilla of Bombyx mori was studied in cryofixed specimens. These sensilla belong to the category of double-walled wall-pore sensilla. The pegs are approximately 10 mum long, located in pits on the dorsal side of the antennal branches, and longitudinally grooved in their distal half (grooved surface approximately 30 mum(2)). The central lumen contains the outer dendritic segments of usually five receptor cells, and is surrounded by up to 15 partially fused cuticular fingers. The peripheral lumina of these cuticular fingers are filled with material resembling wax-canal filaments. Radial spoke channels ( approximately 600 per peg), each 10-20 nm wide, connect the central lumen with the longitudinal groove channels. Groove and spoke channels are assumed to mediate the transport of odorant molecules from the outer epicuticular surface layers to the sensory dendrites. Thus the double-walled wall-pore sensilla represent a bauplan essentially different from single-walled wall-pore sensilla; the reason, however, why the two types are found together throughout the insect orders remains enigmatic. Other peculiar features of the coeloconic sensilla of the silkmoth are invaginations of the outer dendritic segments and direct contacts between the receptor cell somata. The latter may be the structural correlate to electrophysiological observations indicative of peripheral interaction between the receptor neurons. All three auxiliary cells have elaborately folded apical plasma membranes studded with portasomes and associated with an abundance of mitochondria; basally they often contact tracheal branches. As compared to the auxiliary cells of the single-walled olfactory sensilla of the same species, all the mentioned features are much more prominent and hint to a higher ion pumping activity at the border to the sensillum-lymph cavities.     

The statocyst of Vampyroteuthis infernalis (Mollusca: Cephalopoda)

The statocyst shows a remarkable combination of features of decapods and octopods confirming that Vampyroteuthis is a relic somewhere near the ancestor of both groups. The lining of the statocyst separates from the outer wall, forming an inner sac, filled with endolymph, surrounded by perilymph. This is the condition found in octopods, never in decapods. The macula is partly divided into a macula princeps and macula neglecta, as in decapods but never in octopods. There are numerous statoconia, but no large statolith has been seen. The crista has four parts as in decapods, but they are not sharply separated. There are numerous small anticristae, with the general form found in decapods, differentiated into pegs and hooks.
The wall of the inner sac contains numerous hair cells. These hairs protrude between the epithelial cells. The bases of the cells are drawn out into fine processes, presumably some dendritic and some axonal. There is thus a plexus of nerve fibres all over the wall, communicating with the crista nerve.
There is a very large posterior sac of unknown function, lying behind the crista. It contains only one large anticrista and the opening of Kölliker's canal, which is very large.