Design of an insect cuticle associated with osmoregulation: the porous plates of chloride cells in a mayfly nymph.

In mayfly nymphs of the genus Coloburiscoides, cell complexes with an osmoregulatory function (so-called chloride cells) are found in the integuments of the oral gills, the abdominal gills and gill filaments, the coxae and the thoracic sternites. The cuticle overlying each cell complex is a rigid circular plate which is known to be porous to colloidal lanthanum suspensions. The present study shows that the plate is composed only of the cuticulin and dense layers of the epicuticle. Both layers have substructures built of subunits on almost perfect hexagonal lattices. The lattice spacings are 53 and 9.5 nm for the dense layer and the cuticulin layer respectively. During moulting the apical plasma membrane of the chloride cell remains adpressed to the old porous plate. The new porous plate is formed from a new chloride cell which intrudes from the base of the integument. Throughout the moult small pores persist in the new and otherwise continuous cuticle to allow continuity of the cytoplasm of the apical and basal portions of the old chloride cell. It is thought that this phenomenon allows osmoregulatory function of the chloride cell complex to be maintained during the moult.

A proposed solution to a fine-structural puzzle: the organization of gill cuticle in a crayfish (Panulirus).

Crayfish gill cuticle is approximately 2 micron thick and comprises an epicuticle and an endocuticle, which is subdivided into outer and inner layers. Sections demonstrate indistinct lamellae in the outer endocuticle and vertically striated lamellae in the inner endocuticle. Microfibrils cannot be seen in sections. Difficulties in interpretation of the fibrous architecture of the cuticle from thin sections have been overcome by examining tilted series of micrographs of sections and also by making freeze-fracture replicas of the cuticle, which reveal the microfibrils clearly. A model for the endocuticle based on a helicoidal configuration of microfibrillar laminae is proposed and the vertically striated structures seen in sections of the outer layer are accounted for by including regular rows of particles oriented perpendicular to microfibrils. The model is compared with cuticles and coverings reported from other invertebrates.

Fine structure of the cuticle of the desert scorpion, Hadrurus arizonensis.

The structure of the sclerite and intersegmental cuticle of the opithosoma of the desert scorpion, Hadrurus arizonensis, has been examined by transmission electron microscopy. The sclerite cuticle contains a four-layered epicuticle, a hyaline exocuticle, an inner exocuticle and an endocuticle. The outer part of the hyaline exocuticle and the whole of the inner exocuticle are constructed of helicoidally arranged planes of microfibrils. Within the endocuticle, the overall architecture is not helicoidal as previously assumed, but consists of bundles of microfibrils oriented horizontally and vertically. Microbibrils of the inner exocuticle and the endocutile are seen as simple unstained rods, but those of the hyaline exocuticle are electron dense rods with an unstained central core. The intersegmental cuticle contains a four-layered epicuticle and a procuticle. In detail, its fine structure differs in most respects from that of the sclerite cuticle. Electron microscopy reveals that hyaline exocuticle, previously assumed to be continuous from sclerite to intersegmental membrane, is absent in the latter.

Fine structure of the epicuticle of the desert scorpion, Hadrurus arizonensis, with reference to location of lipids.

The surface and transverse sections of the epicuticle of the desert scorpion, Hadrurus arizonensis, were examined by scanning and transmission electron microscopy, respectively. Sclerite cuticle that was untreated prior to normal EM preparative procedures was compared to cuticle subjected to lipid solvents, high temperature, and concentrated alkali. Surface morphology of untreated intersegmental cuticle was also examined. The epicuticle is composed of four sublayers: outer membrane, outer epicuticle, cuticulin, and the dense homogeneous layer. Lipid solvents did not significantly alter the morphology of any of these layers or the contents of the wax canals that penetrate the cuticulin layer even though the solvents effectively remove lipids from the epicuticle for chemical analysis. The surface of the sclerite cuticle contains amorphous particles, crystalline projections, and scattered openings to dermal gland ducts. Perforations that correspond to the opening of wax canals were faintly visible after extraction of surface waxes and clearly visible after KOH treatment. No openings to dermal gland ducts or wax canals were observed in untreated intersegmental cuticle. However, wax canals are likely obscured by surface waxes similar to those present in sclerite cuticle.

Junctional structures in hydra.

The sealing and communicating junctions present in hydra have been examined using conventional staining, lanthanum tracer and freeze-fracturing techniques. The presence of distinct types of gap and septate junctions has been confirmed. Combined lanthanum tracer and freeze-fracture results have provided a more detailed understanding of these junctional structures. A model has been constructed which demonstrates the various aspects of the junction seen at different sectioning angles. The probable lengths of septa within septate junctions and the junctional 'maze' formed by them is discussed because of its bearing on the 'sealing' nature of the junction and also, to some extent, on its permeability to tracers such as lanthanum.

Comparative ultrastructural studies of insect granulosis and nuclear polyhedrosis viruses.

Optical diffraction studies indicated that the periodic lattice structure in electron micrographs of the capsids of two granulosis and two nuclear polyhedrosis viruses were indistinguishable. The capsid is composed of stacked rings spaced 4-5 nm apart. Comparison of the intracellular forms of Bombyx mori nuclear polyhedrosis virus with negatively stained virus particles leads to encourage the more general use of the term 'capsid' instead of 'intimate membrane' and the term 'virus membrane' in place of 'developmental membrane'. These terms are consistent with those currently used for most animal and plant viruses.

Goblet cell membrane differentiations in the midgut of a lepidopteran larva.

So-called goblet cells are present in the midgut of lepidopteran larvae. They are thought to be involved in the active transport of potassium out of the haemolymph and into the gut lumen. A number of plasma membrane differentiations within the goblet cell cavity has been investigated using conventional staining, lanthanum tracer and freeze-etch techniques. Of particular interest are junction-like inter- and intra-membrane differentiations found on the villus-like cytoplasmic projections present at the apical tip of the goblet cell cavities. These cytoplasmic projections appear to act as a valve; in some cases they seem to close off the top of the goblet cell cavity, so isolating it from the gut lumen, while in other cases they are spread apart leaving a wide channel from the cavity into the lumen. The junction-like structures on these cytoplasmic projections are different in structure from the septate-type junctions which seal the midgut cells together at their apical borders, and the 2 types are present on the same plasma membrane, often within one micron of each other. The need for a different type of junction may possibly be related to the fact that it occurs between 2 areas of the same plasma membrane. The morphology of this unusual junction-like structure is discussed and 2 diagrams are presented to illustrate our interpretation of its structure.

Scanning and transmission electron microscopy of the spermiophores of Ornithodoros ticks: an attempt to explain their motility.

The spermiophores of two tick species, the kangaroo tick, Ornithodoros gurneyi and the cave tick, Ornithodoros tholozani have been examined by scanning and transmission electron microscopy. The anterior end (head) of the spermiophore is a hemisphere covered with a hexagonal network of small projections. The rest of the spermiophore is covered with longitudinal ridges, seen in sections as cellular processes whose membranes are attached only at their anterior ends by specialized 'feet'. In the cytoplasm of the sperm cell body and just beneath the cellular processes are find filaments, which form a continuous layer in O. tholozani and bundles in O. gurneyi. Fibrils tend to be situated beneath the larger cellular processes. In scanning micrographs helical constrictions have been observed in the posterior parts of some spermiophores. It is proposed that certain of the movements observed by light microscopy in living cultures of spermiophores may be explained by contraction of the cytoplasmic filaments seen in the electron microscope.

Junctional structures in the midgut cells of lepidopteran caterpillars.

The junctional structures present between the midgut cells of 3 lepidopteran caterpillars have been examined using freeze-etching, conventional staining and lanthanum tracer techniques. The bonding junction present in this type of tissue is the so-called continuous junction. Septa have only occassionally been reported in conventionally strained cross-sections of these junctions. During the present study septa have been observed in such sections but were more readily located in tissue treated with lanthanum tracer. Tangential sections of lanthanum-impregnated tissue show that these septa are parallel-sided, in contrast to the honeycomb appearance of the septa in septate junctions. The septa in freeze-etch replicas of glutaraldehyde-fixed tissue often show continuous rods on one membrane face, suggesting that the continuous junction may be more akin to the tight junction than to the normal septate junction. However, freeze-etch replicas of unfixed tissue appear much more like replicas of normal septate junctions. The main differences between septate and continuous junctions appear to be that the inter-membrane septa of the continuous junction are parallel-sided in tangential section, as against the honeycomb appearance of the septate junction, and that the particles which delineate the septa in freeze-etched preparations appear to be both somewhat differentl bonded and closer together in the continuous junction. A diagram is presented showing the internal and inter-membrane structures of the 2 types of junction based on the present study, and suggesting a possible explanation of the fact that septa are more readily seen in cross-sections of septate junctions than in continuous junctions. As septa are present in both types of junction, and because their freeze-etch appearances are not very different, it is suggested that the two types of septate junction be called 'smooth septate' and 'pleated septate' junctions, to indicate their characteristic appearances in tangential section.

Hormonal control of morphogenetic cell death of the wing hypodermis in Lucilia cuprina.

1. Cytoplasmic fragments in the haemolymph of newly emerged flies derive from the degenerating wing hypodermis. 2. At the time of eclosion, dorsal and ventral cell layers of the wing are connected by processes containing bundles of microtubules and microfilaments. Cytoplasmic fragments contain similar bundles of microtubules but few microfilaments. 3. Extensive vacuolation marks the onset of hormonally initiated fragmentation of the wing hypodermal cells. Haemocytes containing lysosomes are present in the wing at this time, but do not invade the fragmenting hypodermis.

Ultrastructure of the copper-accumulating region of the Drosophila larval midgut.

The copper-accumulating region of the midgut is a mosaic of interstitial and cup-shaped, copper-accumulating cells. The cup of each cuprophilic cell is lined with a highly refractile border of long microvilli except in one strain where it is predominantly lamellar. The nucleus lies basally; the basal plasma membrane is fairly extensively infolded. Cytolysomes are abundant and increase in number with increasing copper content of the diet. The interstitial cells bear short, less regular microvilli and have a less electron-dense cytoplasm. The nucleus is apical, the mitochondria-associated basal membrane is very extensively infolded and cytolysomes are less abundant. Virus-like particles present in nuclei of both cell types increase in number with increasing copper concentration.

The resistance of epicuticular components of an insect to extraction with lipid solvents.

The cuticle of the Calpodes caterpillar has been treated with lipid solvents and acids prior to examination in the electron microscope. The 'oriented lipid layer', the 'cuticulin layer' and the 'wax canal filaments' are resistant to the action of lipid solvents. The 'cuticulin layer' and the 'oriented lipid layer' are resistant to acids. Terminology of epicuticular structures is discussed and two changes are suggested firstly, to provide a nomenclature that can be applied to homologous structures in the cuticles of other insect species and secondly, to remove ambiguities arising from the results of this investigation.

The fine structure and deposition of the larval cuticle of the sheep blowfly (Lucilia Cuprina).

The cuticle of Lucilia is composed of an untanned endocuticle and a complex epicuticle of four layers, superficial layer, outer epicuticle, cuticulin and dense layer. The outer epicuticle and attached epicuticular filaments are resistant to acid hydrolysis. During deposition of the cuticle of each larval instar, the cuticulin and dense layers are formed first, followed by the outer epicuticle, which appears to be laid down by secretions from the epidermis passing through the cuticulin via epicuticular filaments. The outer epicuticle is found in the position normally occupied by the wax layer of other insect species.

The structure and development of a surface pattern on the cuticle of the green vegetable bug Nezara viridula.

Adult Nezara possess an area of cuticle on the metathoracic sternite with a surface pattern of minute mushroom-like projections. The surface pattern is formed by two types of cell, one forming projections and the other depressions. The surface pattern is largely determined before epicuticle deposition by deformation of the epidermal surface. This is brought about by interactions between the moulting fluid secretions and the cells. Maintenance of shape of the cells may be associated with the presence of oriented cytoplasmic microtubules. The mushroom area may function as an external reservoir causing retarded evaporation of defensive secretions.