Ultrastructural reconstruction of Taenia ovis oncospheres from serial sections

The cellular organisation of Taenia ovis oncospheres is interpreted from ultrathin serial sections and transmission electron microscopy following high pressure freezing and freeze-substitution. The surface of a hatched, non-activated T. ovis oncosphere is covered by an oncospheral membrane below which is the tegument bearing microvilli. The basal lamina of the tegument is underlain by broad bands of peripheral somatic musculature. Three pairs of hooks and associated muscles are present in the somatophoric third of the oncosphere. Approximately 19 cells of seven different types were identified which include: (i) a quadri-nucleated syncytium of penetration gland type 1 containing two lateral pairs of cell bodies interconnected by narrow cytoplasmic bridges (PG1); (ii) a quadri-nucleated syncytium of penetration gland type 2 (PG2); (iii) a single-nucleated median mesophoric gland cell; (iv) 10 somatic cells; (v) two germinative cells; (vi) two nerve cells; and (vii) a pair of median somatophoric cells. This study provides a clear understanding of the morphology of T. ovis oncospheres and forms the basis for further investigations into the biology of taeniid oncospheres.     

Cellular organization of the oncosphere of Mosgovoyia ctenoides (Cestoda: Anoplocephalidae)

The ultrastructure of the infective oncosphere of the cestode Mosgovoyia ctenoides (Anoplocephalidae) is described. The surface of the infective oncosphere is covered by a thin cytoplasmic layer of tegument connected by a narrow cytoplasmic process with the binucleate subtegumental cell, situated deeper in the body. Below the basal matrix of the cytoplasmic layer of the tegument are situated wide bands of the peripheral, somatic musculature responsible for body movements. The 3 pairs of hooks and their muscles form a complex hook muscle system, responsible for coordinated hook action. Five major types of cells have been distinguished: (1) a binucleate subtegumental cell, (2) a binucleate penetration gland, (3) 2 nerve cells, (4) numerous somatic cells, and (5) about 6 germinative cells. The approximate number of cells is 24 (26 nuclei, including 2 syncytial structures). The results of this study, when compared with other published reports from other cestode taxa, support previous hypotheses that the progressive reduction of oncosphere cells is an adaptive feature in cestode evolution.     

Echinococcus granulosus: Hook-muscle systems and cellular organisation of infective oncospheres

The morphology of the oncospheres of Echinococcus granulosus has been reconstructed from thin and semi-thin serial sections. Four major types of oncospheral cells have been identified. These consist of: (1) a bi-nucleate medullary center, (2) glandular regions (eight nuclei) subdivided into three types of oncospheral glands, (3) ten germinative cells, and (4) 34 muscle cells, of which 16 are somatic and 18 are hook muscle cells. The hook muscle cells of each hook are organized functionally into the three following systems: (1) the protraction system, for hook extension, (2) the abduction system for drawing the hooks together toward the median plane of bilateral symmetry, and (3) the retraction system for pulling the hooks back into the body. The interconnections observed between different muscle fibers provide a structural basis for coordinated hook action.     

Cells producing insulin-like androgenic gland hormone of the giant freshwater prawn, Macrobrachium rosenbergii, proliferate following bilateral eyestalk-ablation

We found that the androgenic gland (AG) of Macrobrachium rosenbergii possesses three cell types. Type I cells are small polygonal shaped-cells (13.4 μm in diameter), stain strongly with hematoxylin-eosin (H&;E), have abundant multilayered rough endoplasmic reticulum (rER), and nuclei containing mostly heterochromatin. Type II cells are slightly larger (18.6 μm in diameter), stain lightly with H&;E, have rER with dilated cisternae, and nuclei containing mostly euchromatin. Type III cells (previously undescribed) are similar in size and shape to type I cells, but the cytoplasm is unstained and they have a high amount of smooth endoplasmic reticulum (sER) and mitochondria with tubular cristae. Bilateral eyestalk-ablation resulted in AG hypertrophy with a proliferation and predominance of type I cells as determined by bromodeoxyuridine (BrdU) assays. Expression of insulin-like androgenic gland hormone (Mr-IAG), determined by immunohistochemistry, was weak in type I cells, strong in type II cells of both the intact and eyestalk-ablated, and negative in type III cells. It was also detected in spermatogonia, nurse cells, and epithelium lining of the spermatic duct. The function of Mr-IAG in these tissues is yet to be elucidated but the distribution implies a strong role in male reproduction.     

Comparative ultrastructure of the oesophageal glands of third stage larval hookworms

Smith J. M. 1976. Comparative ultrastructure of the oesophageal glands of third stage larval hookworms. International Journal for Parasitology6: 9–13. The oesophageal glands of the third stage larvae of Necator americanus and Ancylostoma tubaeforme are compared, both before and after penetration through skin. The glands of “infective” larvae of N. americanus are densely packed with secretory granules, contrasting with a reduced gland size in the “penetrated” larvae coupled with the presence of gland secretions in the oesophageal lumen.No difference was observed between the glands of “infective” and “penetrated” larvae of A. tubaeforme. The role of oesophageal gland secretions for penetration of host skin is discussed.     

Extent of polyteny in the pericentric heterochromatin of polytene chromosomes of pseudonurse cells of otu (ovarian tumor) mutants of Drosophila melanogaster

In the polytene nuclei of germ-line cells (ovarian pseudonurse cells) of Drosophila melanogaster females mutant for otu ¹¹ (ovarian tumor), the pericentric heterochromatin is much more abundant than in somatic salivary gland cells. This is due to the degree of heterochromatin compaction (and consequently the level of underreplication) being lower in the nurse cells than in the salivary gland cells. The lower level of compaction probably results in a very low degree of position effect gene inactivation in the ovarian nurse cells.     

Ultrastructure of the salt glands of the mangrove,Avicennia marina (Forssk.) Vierh., as indicated by the use of selective membrane staining

Each salt-excreting gland of the mangrove Avicennia marina (Forsskål) Vierh. consists of two to four collecting cells, one stalk cell, and eight to twelve excretory cells. Differential membrane staining by zinc iodide-osmium tetroxide (as a post-fixative) or phosphotungstic acid (as a section-stain) was used to characterise the ultrastructure of the glands. A large amount of tubular endoplasmic reticulum was found in the stalk and excretory cells of the gland, but not in the collecting cells. The ultrastructural arrangement of the endoplasmic reticulum indicates that salt is loaded from the apoplasm into the endoplasmic reticulum of the symplasm at the base of the stalk cell, traverses both cell types in the endoplasmic reticulum, and is excreted at the outer edge of the gland by an eccrine-type mechanism. Increasing development of the tubular endoplasmic reticulum accompanied differentiation of the gland cells.Abbreviations ER endoplasmic reticulum - PTA phosphotungstic acid - ZIO zinc iodide-osmium tetroxide     

Bilateral eyestalk ablation of the blue swimmer crab, Portunus pelagicus, produces hypertrophy of the androgenic gland and an increase of cells producing insulin-like androgenic gland hormone

The androgenic glands (AG) of male decapod crustaceans produce insulin-like androgenic gland (IAG) hormone that controls male sex differentiation, growth and behavior. Functions of the AG are inhibited by gonad-inhibiting hormone originating from X-organ-sinus gland complex in the eyestalk. The AG, and its interaction with the eyestalk, had not been studied in the blue swimmer crab, Portunus pelagicus, so we investigated the AG structure, and then changes of the AG and IAG-producing cells following eyestalk ablation. The AG of P. pelagicus is a small endrocrine organ ensheathed in a connective tissue and attached to the distal part of spermatic duct and ejaculatory bulb. The gland is composed of several lobules, each containing two major cell types. Type I cells are located near the periphery of each lobule, and distinguished as small globular cells of 5-7 μm in diameter, with nuclei containing mostly heterochromatin. Type II cells are 13-15 μm in diameter, with nuclei containing mostly euchromatin and prominent nucleoli. Both cell types were immunoreactive with anti-IAG. Following bilateral eyestalk ablation, the AG underwent hypertrophy, and at day 8 had increased approximately 3-fold in size. The percentage of type I cells had increased more than twice compared with controls, while type II cells showed a corresponding decrease.     

Seasonal changes in the structure and secretory activity of the androgenic gland of Travancoriana schirnerae

This study investigated the seasonal variation in the structure and secretory activity of the androgenic gland (AG) in the freshwater crab: Travancoriana schirnerae. The androgenic gland is an elongate structure, attached to one side on the wall of the ejaculatory duct. Histological studies showed the presence of three cell types, which differ in size, shape of nuclei, and presence or absence of secretory vesicles. Type I cells are small with large nuclei whereas type II cells are large with small nuclei. Type III cells are intermediate in size and exhibited streak-like nuclei and transparent cytoplasm. Seasonal changes were discerned in the morphology, histology and secretory activity of the gland. March-June appeared to be the active season with type II cells containing secretory vesicles. The mode of release of secretion found to be holocrine. The secretory activity almost completed by July-August (the mating season) with vacuolization of type II cells. The gland remained inactive from September-December with abundance of vacuoles, scattered pycnotic nuclei, indistinct cell membranes and total cellular degeneration. January-February was the revival period with type I cell proliferation. The present study revealed that the secretory activity of the gland is in tune with the male reproductive cycle.     

Schistosoma mansoni: penetration apparatus and epidermis of the miracidium

Free swimming miracidia of Schistosoma mansoni were studied with the electron microscope for the purpose of describing the penetration apparatus and the epidermis. The penetration apparatus was composed of 3 unicellular glands which contain membrane-bound vesicles containing macromolecular diglycols. Each gland contained a nucleus, rough endoplasmic reticulum, Golgi complexes, numerous mitochondria, and glycogen stores. Each gland cell opened to the exterior through the apical papilla.The surface of the miracidium, with the exception of the apical papilla, was covered with ciliated epidermal cells containing numerous mitochondria, membranous bodies, and glycogen. No nuclei were detected within these epidermal cells. Intercellular ridges connecting with subepidermal cytons interrupted the epidermal cells at numerous points. The ridges were joined to the epidermal cells by septate desmosomes. Beneath the epidermal cells were found circular and longitudinal muscle bundles.Sensory structures of various types were associated with the outer covering. These consisted of (1) numerous “knob-like” cytoplasmic projections associated with epidermal cells, (2) bulbous, lamelloid structures with external cytoplasmic projections, and (3) ciliated nerve endings with posterior epidermal tiers and ciliated nerve pits associated with apical papilla.     

Tissue-Specific Differences in the Spatial Interposition of X-Chromosome and 3R Chromosome Regions in the Malaria Mosquito Anopheles messeae Fall.

Spatial organization of a chromosome in a nucleus is very important in biology but many aspects of it are still generally unresolved. We focused on tissue-specific features of chromosome architecture in closely related malaria mosquitoes, which have essential inter-specific differences in polytene chromosome attachments in nurse cells. We showed that the region responsible for X-chromosome attachment interacts with nuclear lamina stronger in nurse cells, then in salivary glands cells in Anopheles messeae Fall. The inter-tissue differences were demonstrated more convincingly in an experiment of two distinct chromosomes interposition in the nucleus space of cells from four tissues. Microdissected DNA-probes from nurse cells X-chromosome (2BC) and 3R chromosomes (32D) attachment regions were hybridized with intact nuclei of nurse cells, salivary gland cells, follicle epithelium cells and imaginal disсs cells in 3D-FISH experiments. We showed that only salivary gland cells and follicle epithelium cells have no statistical differences in the interposition of 2BC and 32D. Generally, the X-chromosome and 3R chromosome are located closer to each other in cells of the somatic system in comparison with nurse cells on average. The imaginal disсs cell nuclei have an intermediate arrangement of chromosome interposition, similar to other somatic cells and nurse cells. In spite of species-specific chromosome attachments there are no differences in interposition of nurse cells chromosomes in An. messeae and An. atroparvus Thiel. Nurse cells have an unusual chromosome arrangement without a chromocenter, which could be due to the special mission of generative system cells in ontogenesis and evolution.     

Anatomy and fine structure of pedicellar glands in phoretic deutonymphs of uropodid mites (Acari: Mesostigmata)

Phoretic deutonymphs of uropodid mites are attached to their carrier via an anal pedicel which is formed by a secretion from the pedicellar gland. Since the ultrastructure of the pedicel and pedicellar gland has never been investigated, we studied these structures in three species, Uropoda orbicularis (Müller), Uroobovella marginata (Koch), and Trichouropoda ovalis (Koch) by light (LM) and electron microscopy (TEM, SEM). In addition, the pedicel in Uroobovella nova (Oudemans) was documented in SEM.The pedicellar gland is a distinct globular structure comprised of three types of secretory cells (A-, B-, and C-types) with apical parts directed towards a junctional zone of postcolon and anal atrium. Secretory cells of the A-type are located dorsally, whereas B-type cells are central, and C-type cells are distributed ventrally or both ventrally and dorsally. Protrusions of visceral muscle cells are distributed on the external surface of the gland. The cuticle-lined anal atrium is large and located between the gland and dorso-anal muscles. The pedicel is composed of a main stalk and two extended extremities: one adhering to the anal region of the deutonymph and a second connected to the cuticle of the carrier. In each case, the anal atria were empty, whereas the pedicellar material was located outside of the mite body.     

Microtubule organization and cell division in embryogenie protoplast cultures of white spruce (Picea glauca)

Summary Immunofluorescence methods were developed for examining the distribution of microtubules in freshly isolated and cultured protoplasts and regenerated somatic embryos of white spruce (Picea glauca). Freshly isolated protoplasts consisted of both uniand multinucleate types. Uninucleate protoplasts established parallel cortical microtubules during cell wall formation and cell shaping, divided within 24 h and developed into somatic embryos in culture. Dividing cells were characterized by preprophase bands (PPBs) of microtubules, atypical spindle microtubules focused at the poles and a typical phragmoplast at telophase. Multinucleate protoplasts also established parallel arrays of cortical microtubules during cell wall formation. In addition their nuclei divided synchronously within 4 days, then cell walls formed between the daughter nuclei. Individual multinucleate protoplast-derived colonies subsequently gave rise to elongate suspensor cells thereby forming embryo-like structures by 7 days.     

Mastophorus muris (Nematoda: Spirurina): Ultrastructure of somatic muscle development

Hamada G. S. and Wertheim G. 1978. Mastophorus muris (Nematoda: Spirurina): ultrastructure of somatic muscle development. International Journal for Parasitology8; 405–414. The ultrastructure of the somatic muscle cells of the adult and six developmental stages of Mastophorus were studied. In all stages the cells consisted of a contractile region containing myofibrils separated by dense bands and a noncontractile region with nuclei, mitochondria, glycogen, lipid droplets and vesicles. Two sizes of myofilaments were present. The dense band contained T tubules and sarcoplasmic reticulum, and, in more advanced stages, support filaments, glycogen and dense bodies. The contractile region of the adult muscle cell consisted of several hundred irregularly shaped myofibrils arranged in a random pattern. This pattern of myofibrils was defined as irregular-coelomyarian. The third stage larva had a shallow-coelomyarian myofibril configuration, which changed to coelomyarian in the late third stage through the addition of new myofibrils at the apical contractile border. In the fourth stage larvae, the subdivision of existing myofibrils changed the pattern to irregular-coelomyarian.     

Ultrastructure of the Anterior Body Region of Marine Nematode Deontostoma californicum

The ultrastructure of the anterior body region of the free-living marine nematode Deontostoma calilornicum was studied by electron microscopy. The body wall consists of a nine-layered cuticle, a cellular hypodermis containing eight nerve bundles, and a well-developed coelomyarian somatic musculature. Nerves in the dorsal, lateral, ventral, and submedian hypodermal chords anterior to the nerve ring were observed with regularity. Structure of subventral somatic setae suggests a mechanoreceptive function. The esophagus is cellular and consists of three marginal cells alternating with an equal number of radial muscle cells, three esophageal glands, and three enteric nerves. The membranes of adjacent esophageal cells are sinuous. Apices of the triradiate lumen are connected with the outer wall of each marginal cell by bands of electron-dense nonmyofibrils, whereas two types of myofilaments run radially between the apophyses of the lumen and the outer walls of radial cells. Each myofibril, which forms hemidesmosomes at both ends, is interpreted to be the morphological equivalent of one sarcomere. Synaptic junctions between the processes of muscles, gland cells, and axons of the enteric nerves are described in detail.     

Ultrastructural localization of egg-laying prohormone-related peptides in the atrial gland of Aplysia californica

The atrial gland is an exocrine organ that secretes into the oviduct of Aplysia californica and expresses three homologous genes belonging to the egglaying hormone gene family. Although post-translational processing of the egg-laying hormone precursor in the neuroendocrine bag cells has been examined in detail, relatively little is known about the post-translational processing of egg-laying hormone-related gene products in the atrial gland. A combination of morphologic techniques that included light-microscopic histology and immunocytochemistry, transmission electron microscopy, and immuno-electron microscopy were used to localize egg-laying hormone-related peptides in the atrial gland and to evaluate the characteristic morphology of their secretory cells. Results of these studies showed that there were at least three major types of secretory cells in the atrial gland (types 1–3). Significantly, of these three cell types, only type 1 was immunoreactive to antisera against egg-laying hormone-related precursor peptides. The immunoreactivity studies established that all three egg-laying hormone-related precursor genes are expressed in type-1 cells and indicated that the processing of these precursors also occurs within the secretory granules of this cell type. Evidence was also obtained that proteolytic processing of the egg-laying hormone-related precursors differed significantly from that observed in the bag cells. In contrast to the bag cells, the NH₂-terminal and COOH-terminal products of the egg-laying hormone-related precursors of the atrial gland were not sorted into different types of vesicles.     

Schistosoma mansoni: Ultrastructure of early transformation of skin- and shear-pressure-derived schistosomules

Against the background of cercarial fine structure, ultrastructural changes were compared in schistosomules of Schistosoma mansoni 30 min and 1 hr after their production in vivo by skin penetration and in vitro by shear pressure. The same developmental pattern was observed in schistosomules of both derivations. In vitro schistosomules, however, developed more slowly, resembled cercariae more closely, and varied less among organisms than did in vivo schistosomules. The greatest morphological changes were observed in the 1-hr in vivo schistosomules. These were as follows: (1) in tegument, formation of transient microvilli, a hepatalaminate outer membrane and accented surface invaginations, loss of glycocalyx, movement outward of cyton vesicles via bridges, accumulation of multilaminate bodies around bridge openings; (2) in the anterior organ (oral sucker), movement of head gland vesicles via the ducts into tegument followed by collapse of the gland fundus, disappearance of the circumfundal cells and two large support cells, and the appearance in these areas of membranes and parenchymal cells; (3) secretion of the acetabular gland contents, collapse of the glands and replacement by membranes and parenchymal cells; (4) peristaltic activity of the digestive tract as shown by alternate areas of lumen constriction and dilation; (5) loss of bladder and contraction of the small aboral collecting tubules; and (6) conversion of heterochromatic parenchymal cell nuclei to euchromatic. In contrast, the 1-hr in vitro shear schistosomules resembled 30-min in vivo schistosomules, retaining many cercarial features.     

Co-expressed Cyclin D variants cooperate to regulate proliferation of germline nuclei in a syncytium

The role of the G1-phase Cyclin D-CDK 4/6 regulatory module in linking germline stem cell (GSC) proliferation to nutrition is evolutionarily variable. In invertebrate Drosophila and C. elegans GSC models, G1 is nearly absent and Cyclin E is expressed throughout the cell cycle, whereas vertebrate spermatogonial stem cells have a distinct G1 and Cyclin D1 plays an important role in GSC renewal. In the invertebrate, chordate, Oikopleura, where germline nuclei proliferate asynchronously in a syncytium, we show a distinct G1-phase in which 2 Cyclin D variants are co-expressed. Cyclin Dd, present in both somatic endocycling cells and the germline, localized to germline nuclei during G1 before declining at G1/S. Cyclin Db, restricted to the germline, remained cytoplasmic, co-localizing in foci with the Cyclin-dependent Kinase Inhibitor, CKIa. These foci showed a preferential spatial distribution adjacent to syncytial germline nuclei at G1/S. During nutrient-restricted growth arrest, upregulated CKIa accumulated in arrested somatic endoreduplicative nuclei but did not do so in germline nuclei. In the latter context, Cyclin Dd levels gradually decreased. In contrast, the Cyclin Dbβ splice variant, lacking the Rb-interaction domain and phosphodegron, was specifically upregulated and the number of cytoplasmic foci containing this variant increased. This upregulation was dependent on stress response MAPK p38 signaling. We conclude that under favorable conditions, Cyclin Dbβ-CDK6 sequesters CKIa in the cytoplasm to cooperate with Cyclin Dd-CDK6 in promoting germline nuclear proliferation. Under nutrient-restriction, this sequestration function is enhanced to permit continued, though reduced, cycling of the germline during somatic growth arrest.     

The use ofXenopus oocytes and embryos as a route towards cell replacement

When nuclei of somatic cells are transplanted to enucleated eggs ofXenopus, a complete reprogramming of nuclear function can take place. To identify mechanisms of nuclear reprogramming, somatic nuclei can be transplanted to growing meiotic oocytes ofXenopus, and stem cell genes activated without DNA replication. The combination of somatic cell nuclear transfer with morphogen signalling and the community effect may lead towards the possibility of cell replacement therapy. When mechanisms of nuclear reprogramming are understood, it may eventually be possible to directly reprogramme human somatic cell nuclei without the use of eggs.