Intraspecific variation in the terrestrial isopod Oniscus asellus Linnaeus, 1758 (Crustacea: Isopoda: Oniscidea)

A study of intraspecific variation in one of the most widespread and abundant terrestrial isopods in Europe, Oniscus asellus , is presented. It is concluded that previous attempts to describe intraspecific taxa in this species are invalid, but that the species is clearly divisible into two subspecies which differ in a number of features, including male sexual modifications. The nominal subspecies, Oniscus asellus asellus Linnaeus, 1758, is described in detail, and a neotype, collected from Uppsala, Sweden, where Linnaeus lived and worked, is designated. Oniscus asellus occidentalis ssp. nov . is also described, and reported as occurring in the far west of the species' range, in the British Isles, Ireland and France. Characters useful in distinguishing these two taxa are indicated and figured. Differences in the ecology of the two taxa are also noted, occidentalis appearing to be a relict Atlantic taxon, largely restricted to damp woodland. The existence of populations intermediate between asellus and occidentalis is noted, and the features of these populations discussed in detail. A combination of geographical, environmental and laboratory breeding data suggest that these intermediates arise through hybridization between the two subspecies, this process of hybridization being accelerated by human activity, which brings the two taxa into frequent contact. It is suggested that Oniscus asellus may provide an excellent model system for the study of the dynamics of hybrid zones.     

Nerve terminals and epithelial cell variety in the human lacrimal gland

Summary Pieces of tissue taken from three human lacrimal glands were examined electron microscopically. Secretory cells varied according to the electron density and structure of their secretion granules. Secretory cells were arbitrarily categorized as light, medium or dark based on their granule content. Acini were composed of two or all three categories of cells together with myoepithelial cells and lymphocytes.A minority of interstitial nerve fibres contained small granular vesicles characteristic of adrenergic, sympathetic terminals. The appearance of most interstitial and all parenchymal fibres was consistent with that of cholinergic, parasympathetic terminals. Parenchymal terminals were confined to ducts, terminal tubule areas and to serous (dark) cells. A large proportion of parenchymal terminals lay adjacent to myoepithelial cells in the ducts and terminal tubule regions but terminals observed among serous cells were rarely in contact with myoepithelial cells. A possible parasympathetic control of serous secretion, granule production and duct contraction and the autonomy of mucous cells is discussed.The author is indebted to Dr. J. Egeberg of the Anatomy Department B, University of Copenhagen for the provision of embedded material.     

Ultrastructure and nature of secretory proteins in the male accessory gland of Drosophila funebris

The ultrastructural differentiation of the secretory cells and the nature of secretory proteins in the male accessory gland of Drosophila funebris have been studied by electron-microscopic and immunological methods. (1) In the pupae at $\text{1}\text{1}\text{2}$ days before eclosion, secretory products can be detected in the lumen, even though most glandular cells are at the initial phase of differentiation. At the time of eclosion both main and secondary cells are fully differentiated, but the whole set of five immunologically active proteins are detectable only on the second to third day of adult life. (2) The secondary cells contain giant protein granules, the so-called filamentous bodies, which become partially fused and the filaments assume a twisted form. Randomly dispersed filaments and closely packed filament bundles are also visible in the gland lumen. Antigenic labelling of ultrathin sections and immunoreplica electrophoresis yielded no evidence for the microtubular nature of these filaments. The secretion stored in the lumen contains in addition a large quantity of flocculent proteins which have their origin in the main cells. (3) During the period of high secretory activity in the 7-day-old male flies no vacuolization and disintegration of either the main or secondary cells have been observed. We conclude that both types of cells have the merocrine secretory mechanism. (4) Ultrastructural alterations in the glandular cells confirmed our previous observation that copulation stimulates RNA and protein synthesis.     

The ultrastructure of the heart of Oniscus asellus L. and Asellus aquaticus L. (Crustacea,Isopoda)

Summary The endocardium of Oniscus asellus L. and Asellus aquaticus L. consists of lipid cells. The epicardium consists of a layer of cells with a vesiculated cytoplasm covered by a thick extracellular fibrous sheet. The myocardium is a single layer of cells, the sarcolemma invaginates at Z disc level forming transverse tubules, and longitudinal tubules branch off from these. At the A-I level' longitudinal tubules form transverse systems, which form couplings with the sarcoplasmic reticulum. The sarcoplasmic reticulum appears as perforated sheets enveloping the myofibrils. Two types of nerve terminal are found: one is embedded in a myocardial cell process, the other lies in a myocardial cell depression. They contain clear and dense-cored synaptic vesicles.This work was supported by grants from the Norwegian Research Council for Science and the Humanities     

Ultrastructure of secretion in the atrial gland of a mollusc (Aplysia)

Extracts of the atrial gland of the sea hare Aplysia californiea (Mollusca) induce egg laying when injected into mature individuals. Since egg laying is controlled endogenously by a peptide secreted by neuroendocrine cells in the central nervous system, the relationship between the atrial gland and these central neurons has become an issue of interest. With the particular objective of examining secretory structures we undertook an ultrastructural study of the atrial gland and adjacent tissues. This study revealed that the atrial gland epithelium is composed of two major cell types: ‘goblet-like’ exocrine cells containing large electron-dense granules, and ciliated ‘capping cells’. A non-secretory, and possibly post-secretory, cell containing electron-lucent granules was noted. A region of the large hermaphroditic duct contiguous to the atrial gland, known as the red hemiduct, also displayed capping cells and secretory cells with large granules. The content of these granules is organized into crista-like condensations. The cell also contains iron-rich pigment inclusions.     

A cryofixation study of presumptive hygroreceptors on the antennule of a terrestrial isopod

The structure of the apical sensilla on the antennule of the terrestrial isopod Porcellio scaber was examined in cryofixed and freeze substituted (CRF) and chemically fixed and dehydrated (CHF) material. CRF specimens generally showed a preservation superior to CHF material. Only in deeper regions did the tissue show damage from freezing. Each of the 13–22 sensilla contains two sensory cells. In contrast to earlier reports, it was observed that the dendritic segments of these cells are arranged in a unique, concentric manner. In CRF specimens the dendrites reach the tip of the sensilla and border upon the innermost layer of the complicated wall of the peg which is not pierced by pores. Silver-protein and lanthanum failed to penetrate the wall of the sensilla and also did not reach the dendrites via an apical pore, which therefore is regarded as a molting pore. The lymph spaces which, in CHF specimens, are observed around the dendrites and beneath the cuticle within the antennal tip are regarded as artefactual. From similarities in the dendritic structures to insect hygroreceptors and their relationship to the adjoining cuticle it can be assumed that the antennular sensilla in Porcellio are sensitive to humidity changes. Mechanoreception and chemoreception, however, cannot entirely be excluded as possible functions.     

Dietary effects on life history traits in a terrestrial isopod: the importance of evaluating maternal effects and trade-offs

Studies of life history aim to explain patterns in the evolution of reproductive investment, growth, and survival. Trade-offs between traits are a fundamental component of life history theory. In herbivorous arthropods life history traits are often responsive to variation in numerous environmental factors, especially diet quality. Using three artificial diets under controlled laboratory conditions, we examined changes in life history traits (i.e. growth rate, offspring number, offspring size, incubation period), trade-offs between traits, and maternal effect on the growth rate of offspring, in the common woodlouse (terrestrial isopod), Porcellio laevis. The high protein diet had significant impacts on offspring production, triggering a smaller-sized offspring, and demonstrating a trade-off between these last two traits. The high carbohydrate diet seldom exerted a significant effect on incubation period. The quality of dietary items evidently has important consequences on the life history of the mother and, thus, on offspring growth; the directions of these effects, however, were opposite. Mothers fed diets with high protein concentrations presented significant maternal effects, measured as offspring growth rate during later ontogeny. Our results support the notion that protein, rather than carbohydrate, concentrations in the diet limit herbivorous arthropods, and have significant consequences on life history traits, as was seen for P. laevis. Clearly, the change in phenotypic correlations between incubation period and offspring number from negative to positive is an empirical demonstration of the context dependence of life history trait trade-offs.     

A comparative study on the fine structure of developing spermatozoa in the isopod,Oniscus asellus,and the amphipod,Orchestoidea sp.

Summary Developing spermatids and mature spermatozoa from the isopod, Oniscus asellus and the amphipod, Orchestoidea sp. have been examined with the light microscope and the electron microscope and have been found to have similar morphologies. As spermiogenesis proceeds the nucleus migrates to one pole of the spermatid at which point an acrosome, contiguous rod, and cross-striated tail develop. The acrosomal vesicle elongates to a cone-shaped, mature acrosome lying at the apex of a cross-striated tail and nucleus which are situated at approximate forty-five degrees to each other. The cross-striated tail originates as an evagination of the spermatid plasma membrane near the acrosomal vesicle. The tail eventually grows to lengths of four to five hundred microns. The mature, tail-like appendage is cross-striated at major 750 to 800 Å, and minor 125 to 150 Å, periodicities. When observed in vitro, mature sperm of both species appear non-motile.Possible homologies of this unusual spermatozoon with other types of spermatozoa are made and it is concluded that: 1) isopod and amphipod spermatozoa should be classified as non-flagellate; 2) the cross-striated tail, previously thought to be a flagellum, is a non-motile structure associated in development and possible function with the acrosome; and 3) the rodlike structure contiguous with the acrosome is similar to perforatoria described in some vertebrate sperm.Supported by U.S.P.H.S. Grant No. NB-06285 and Training Grant No. 5-Tl-GM-202. — The author wishes to express his grateful appreciation for the technical assistance given by Miss Ann Barnett during the course of this investigation.     

Heavy metals in the terrestrial isopod Porcellio scaber Latreille. II. Subcellular fractionation of metal-accumulating lysosomes from hepatopancreas

Two populations of metal contaminated Porcellio scaber Latreille were studied: one consisting of animals that had been fed heavy metals in the laboratory for several months, and one from a metal-polluted site in the field (Braubach, FRG). Density gradient centrifugation was performed on hepatopancreas homogenates in order to identify cellular fractions and their association with lead, copper and cadmium. Marker enzymes were used for localisation of cellular fractions in the density gradient. Two lysosomal fractions, called the light and heavy fraction, were separated. They contained mainly lead, but also copper and some cadmium.Abbreviations AP acid phosphatase - ER endoplasmic reticulum - LDH lactic dehydrogenase - MDH malic dehydrogenase - NADH nicotinamide adenine dinucleotide - P. scaber Porcellio scaber     

Dietary effects on growth, reproduction, body composition and stress resistance in the terrestrial isopods Oniscus asellus and Porcellio scaber

The effect of differences in food composition on growth, body composition and cold tolerance of the isopods Porcellio scaber (Latreille) and Oniscus asellus (L.) has been studied. The effect on reproduction of P. scaber was included. Total lipid and protein content remained the same in O. asellus, given food with a nitrogen content of 2.4, 2.5, 2.8 or 3.1% for 8 weeks. Those fed on 3.1% N gained significantly more weight than those fed on 2.4, 2.5 or 2.8% N. Cold tolerance after a period of low temperature acclimation could not be tested, due to high mortality at the subzero treatment. Haemolymph osmolality increased after the winter acclimation for all food treatments. Juvenile P. scaber, given food of 1.8, 2.5 or 3.1% N, showed no difference in growth over 18 weeks, nor in body composition over 14 weeks. Diet did not influence cold tolerance ability and haemolymph osmolality remained the same with changing cold tolerance of the animals. Gravid females of P. scaber were separated to examine the effect of diet on reproduction. Diet did not influence the number of juveniles, nor their weight. In all food treatments, a significant correlation was found between the weight of the female and the number and weight of her offspring.     

Ultrastructural study of the neurosecretory granules in the sinus gland of the blue crab,Callinectes sapidus

Summary Seven morphologically different types of neurosecretory granules have been found in the axon terminals of the sinus gland of the blue crab, Callinectes sapidus. They differ from each other in size, shape, staining characteristics, solubility characteristics, core matrix characteristics, axon terminal matrix characteristics, presence or absence of space between the granule membrane and granule core matrix, and frequency of occurrence. Five of the types are segregated in different axon terminals and are believed to represent different hormone-protein complexes. Two of the types, which have lost part or all of their granular contents, are thought to be variants of the other five types. The differences in granular morphology are better revealed by some fixation procedures than others. Palade's acetate-veronal buffered osmium tetroxide, in particular, reveals striking differences. The following observations suggest that different hormone-protein complexes are segregated in different axon terminals and that these complexes may be morphologically distinguished at the level of the electron microscope.Supported by USPHS-NIH Training Grant GM-00669 and Grant GB-7595X from the National Science Foundation.     

Ultrastructural appearance of neurosecretory granules in the sinus gland of the crab after different fixation procedures

Summary The appearance of neurosecretory granules in the crab sinus gland was studied after fixation at different pHs. Whereas at pH 7.0 the neurosecretory granules were pleomorphic with respect to electron density, at pH 5.0 or 6.0 all the granules remained electron dense. The possible role of maturation as an explanation of this observation is discussed.ERA 493 CNRS     

Heavy metals in the terrestrial isopod Porcellio scaber latreille. I. Histochemical and ultrastructural characterization of metal-containing lysosomes

Different populations of metal-loaded and uncontaminated Porcelio scaber Latreille were studied. Combined light and electron microscopial methods as well as X-ray microanalysis were applied for localization and characterization of intracellular sites of metal deposition within the small cells of hepatopancreas. By means of cytochemistry and X-ray microanalysis it was shown that membrane-limited vesicles are important sites of deposition for lead, copper, zinc and probably smaller amounts of cadmium. The vesicles also contained phosphorus. They are identical with the reported cuprosomes and belong to the lysosomal system. In addition, considerable amounts of lead, copper, and cadmium were found in small structures outside of membrane-limited organelles.Abbreviations EDX energy-dispersive X-ray - P. scaber Porcellio scaber - STEM scanning transmission electron microscope - TEM transmission electron microscope - v volume - wt weight     

Ultrastructure and ATPase activity of the rectal gland of the chondrichthyean fish Hydrolagus colliei (Holocephali)

Summary The anatomy, histology, ultrastructure and ATPase activity of the intramural rectal gland of the chondrichthyean Hydrolagus colliei, are described. The cells of the rectal gland of Hydrolagus demonstrate the same well developed lateral and basal cisternae, elongate mitochondria and luminal border as those of their elasmobranch counterparts. ATPase activity within the rectal gland of Hydrolagus is as intense as that in a number of elasmobranchs examined in the course of the study. Despite its primitive intramural location the rectal gland of Hydrolagus respresents a homolog of the more specialized and better known elasmobranch gland and appears as well suited for cation excretion.     

Exocytosis of neurosecretory granules from the crustacean sinus gland in freeze-fracture

Exocytosis is clearly shown in freeze-fracture preparations to be the mechanism for neurosecretion granule release from axon endings in the crayfish sinus gland. The cytoplasmic leaflet (A-face) of axon ending membrane is characterized by randomly situated depressions representing invaginations of the axolemma, which are in contact with limiting membranes of neurohormone granules in the subjacent cytoplasm. The extracellular leaflet (B-face) of the axolemma at release sites exhibits complementary volcano-shaped protrusions which are cross-fractures through necks of channels formed by invaginating plasma membrane in contact with underlying neurosecretion granules. Structural variation in B-face protrusions is consistent with a spectrum of exocytotic profiles in various stages of formation, and with granules at different stages of passage out of the endings. Evidence in this study suggests that formation of exocytotic structures may begin by alteration of axon membrane structure at the neurosecretory ending-hemolymph interface prior to contact of the neurohormone granules with the axolemma. Limiting membranes of neurosecretory granules exhibit protrusions which appear to interconnect granules adjacent to release sites and to attach granules to the axolemma. Freeze-fracture is clearly shown to be an invaluable tool for monitoring the degree of exocytosis exhibited by sinus glands under normal conditions and under experimental acceleration of hormone release. This technique is capable therefore, of detecting slight increases in numbers of exocytotic profiles much more quickly and accurately than the examination of random thin sections.     

The distribution of zinc,cadmium, lead and copper within the woodlouse Oniscus asellus (Crustacea,Isopoda)

Summary The concentrations of zinc, cadmium, lead and copper have been determined in the hepatopancreas, hindgut and rest of the body tissues of Oniscus asellus collected from eight sites in the U.K. The hepatopancreas is by far the most important storage organ of heavy metals, particularly cadmium, and at each site, contains a mean of at least 89% of the total body load of this element. Specimens of Oniscus asellus from contaminated sites may contain concentrations of zinc, cadmium, lead and copper in the hepatopancreas of about 1%, 0.5%, 2.5% and 3% of the dry weight respectively, which are among the highest so far recorded in the soft tissues of any animal.There is a significant positive correlation between the mean relative dry weight of the hepatopancreas of Oniscus asellus and the concentrations of zinc or cadmium in leaf litter from all eight sites. It is suggested that animals from sites which are contaminated heavily with zinc or cadmium have a large hepatopancreas because this enables them to de-toxify a greater amount of the metal.