Description of Trichotokara nothriae n. gen. et sp. (Apicomplexa, Lecudinidae) - An intestinal gregarine of Nothria conchylega (Polychaeta, Onuphidae)

The trophozoites of a novel gregarine apicomplexan, Trichotokara nothriae n. gen. et sp., were isolated and characterized from the intestines of the onuphid tubeworm Nothria conchylega (Polychaeta), collected at 20 m depth from the North-eastern Pacific Coast. The trophozoites were 50-155 μm long with a mid-cell indentation that formed two prominent bulges (anterior bulge, 14-48 μm wide; posterior bulge, 15-55 μm wide). Scanning electron microscopy (SEM) demonstrated that approximately 400 densely packed, longitudinal epicytic folds (5 folds/μm) inscribe the surface of the trophozoites, and a prominently elongated mucron (14-60 μm long and 6-12 μm wide) was covered with hair-like projections (mean length, 1.97 μm; mean width, 0.2 μm at the base). Although a septum occurred at the junction between the cell proper and the mucron in most trophozoites, light microscopy (LM) demonstrated that the cell proper extended into the core of the mucron as a thin prolongation. A spherical nucleus (8-20 μm) was situated in the middle of the trophozoites, and gamonts underwent caudal syzygy. The small subunit (SSU) rDNA sequence and molecular phylogenetic position of T. nothriae was also characterized. The sequence from this species was the most divergent of all SSU rDNA sequences currently known from gregarines and formed a weakly supported clade with Lecudina polymorpha, which also possesses densely packed epicyctic folds (3-5 folds/μm) and a prominently elongated mucron.     

The fine structure of the gamont of Pterospora floridiensis (Apicomplexa: Eugregarinida)

Transmission electron microscopy of the gamont stage of Pterospora floridiensis has revealed a number of features. The gamont's surface varies from smooth to crenulate, with numerous pockets and folds. The pellicle is composed of an outer membrane, a middle lucent region, and an inner dense layer comprised of two tightly appressed membranes. Short ridges on the pellicle are 200-300+ nm long, 75-100 nm wide, and have a height of approximately 50 nm. The thickness of the pellicle is 100 nm when measured from the inner membrane to the top of a ridge. The ridges are formed by the plasma membrane and an underlying structure that is circular in cross-section. The surface folds and the pellicular ridges are distributed over the soma and the cell's unusual branching arms, though both are reduced near the junction between two gamonts in syzygy, and are absent at the central area of the junctional site. The cell has numerous active Golgi complexes associated with vesicles, as well as scattered dense mitochondria, lipid droplets, and paraglycogen granules. The nucleus has a large (13 microm) endosome, eccentrically located, and peripheral chromatin along the inner nuclear membrane.     

Observations Biologiques et Ultrastructurales sur les Selenidiidae et Leurs Conséquences sur la Systématique des Grégarinomorphes

SYNOPSIS. The taxonomic position of Selenidiidae Brasil in the class of gregarines is discussed in relation to the study of its life cycles, its possible schizogony and the fine structure of its trophozoites. The cycle of gregarines which belong to the genus Selenidium Giard is characterized by trophozoites with pendular or coiling movements, nuclear transformations in gamonts during syzygy just before cyst formation, anisogamy, sporocysts with 4 sporozoites. Schizogony of the Selenidiidae is not yet demonstrated. The “kystes á meérozoites” within the gut epithelium of Sabellaria alveolata, could be one of the stages of schizogony of S. hollandei. This sole example in our study and the absence of schizogony in numerous species, especially in S. pendula, the type species, shows that this criterion is uncertain, actually, in the definition of the order Archigregarinida Grassé. Ultrastructural studies of S. hollandei and S. pendula show that the cortical region in trophozoites of the genus Selenidium is different from that of Eugregarinida. In the Selenidiidae the epicyte is composed of longitudinal folds. Under the wall, consisting of 3 membranes there is a well-defined pellicular fibrillar system. In S. hollandei, the trophozoite has a fibrillar formation, corresponding to a conoid in its anterior region. The trophozoites of S. hollandei and S. pendula contain anterior dense bodies or rhoptries which are very well developed. All these characteristics conform to the ultrastructural organization of the dissemination forms (merozoites, schizozoites, sporozoites): The results allow one to give a new definition of the Order Archigregarinida: Order Archigregarinida (Grassé): Gregarines with ultrastructural organization of the trophozoites similar to that of dissemination forms. Presence of a well defined pellicular fibrillar system. Intestinal parasites of polychaete worms.     

Evidence for intercellular cohesion in the septate junction of the protozoon Gregarina

Injection of Lucifer Yellow dye was used to establish whether septate junctions formed a permeability barrier between primite and satellite gamonts in the syzygy of the protozoon Gregarina. The fluorescent dye did not pass from one cell to the other, thus suggesting that the septate junction served only for mechanical adhesion. Later on in development, when the gametocyst had formed, the septate junction vanished and interruptions were observed between opposing cell membranes. At this stage the fluorescent dye was able to pass freely into the conjugated cells.     

Gametogenesis and Sporogony of Hepatozoon Mocassini (Apicomplexa: Adeleina: Hepatozoidae) In an Experimental Mosquito Host, Aedes Aegypti

ABSTRACT. The sexual life cycle of the hemogregarine Hepatozoon mocassini was studied in Aedes aegypti , an experimental mosquito host, using transmission electron microscopy. Gamonts were observed leaving the host snake erythrocyte as early as 30 min after mosquitoes ingested infected blood, and some gamonts had penetrated the gut epithelial cells by this time. Six hours post-feeding, gamonts were identified within cells of the abdominal fat body. Twenty-four hours post-feeding, gamonts were often entrapped within the peritrophic membrane, but were no longer observed within the gut wall. Parasites pairing up in syzygy and undergoing sexual differentiatioe were observed within fat cells at this time, and by 48 hours post-feeding, well-developed macro- and microgametocytes as well as microgametes were discernible. Developing zygotes observed 3 days post-feeding were enclosed within a panoitophorous vacuole. By day 6, multinucleate oocysts with crystalloid bodies in the cytoplasm were seen. Sporazoites developing within sporocysts appeared by day 12. Seventeen days post-feeding, mature oocysts with sporocysts containing approximately 16 sporozoites were observed upon dissection of mosquitoes. Large crystalloid bodies no longer bound by rough endopbsmic reticulum were located anterior and posterior to the sporozoite nucleus. Free sporozoites were not observed.     

Morphological changes of sperm nuclei during spermatogenesis in the brown alga Cystoseira hakodatensis (Fucales, Phaeophyceae)

Morphological changes and chromatin condensation of sperm nuclei were observed during spermatogenesis in the fucalean brown alga Cystoseira hakodatensis (Yendo) Fensholt. Ultrastructural studies have shown that the mature spermatozoid has an elongated and concave nucleus with condensed chromatin. The morphological changes and the chromatin condensation process during spermatogenesis was observed. Nuclear size decreased in two stages during spermatogenesis. During the first stage, spherical nuclei decreased in size as they were undergoing meiotic divisions and the subsequent mitoses within the antheridium. During the second stage, the morphological transformation from a spherical into an elongated nucleus occurred. Afterwards, chromatin condensed at the periphery in each nucleus, and chromatin‐free regions were observed in the center of the nucleus. These chromatin‐free regions in the center of nucleus were compressed by the peripheral chromatin‐condensed region. As the result, the elongated and concave nucleus of the mature sperm consisted of uniformly well‐condensed chromatin.     

Electron microscope study of chromatin in hepatocyte nuclei during the first hours after partial hepatectomy. V. Changes in the relative area of condensed chromatin and the density of chromatin fibril packing in the ultrathin sections

The degree of chromatin condensation was studied on ultrathin cell sections of guinea pig hepatocytes during the prereplicative period after partial hepatectomy. Three time points were chosen for analysis namely 2,5, 5 and 9 hrs after operation since they show marked increasing (2.5 hrs), decreasing (5 hrs) and repeated increasing (9 hrs) of the amount of ethidium bromide binding to chromatin. The degree of chromatin condensation was determined by measuring the area occupied by condensed chromatin and also by measuring the number of chromatin fibrils per a certain length. The condensed chromatin with varying localization in the nucleus were studied separately. The changes of nucleoplasmic chromatin were most pronounced: at 2.5 and 9 hrs after operation the decrease of the relative area and of the density of chromatin fibrils package was observed; these parameters were near to control at 5 hrs after operation. In general the changes in nucleoplasmic chromatin were correlated with the changes of the activity of the chromatin in the whole nucleus. The decondensation of the perimembranous chromatin was manifested in the decrease of its area and was expressed only at 9 hrs after operation. The perinucleolar chromatin was found to show the gradual decondensation which was manifested mainly by the decrease of its relative area. Thus the condensed chromatin seems to be a labile structure which undergoes essential changes in the process of the exit of the hepatocytes from G0-stage of the cell cycle, during the prereplicative period.     

The fate of DNA originally existing in the zygote nucleus during achromosomal cleavage of fertilized echinoderm eggs in the presence of aphidicolin: microscopic studies with anti-DNA antibody

In fertilized echinoderm eggs, the male and female pronuclei fuse to form the diploid nucleus even in the presence of aphidicolin, a specific inhibitor of eukaryotic DNA polymerase-alpha. The subsequent first cleavage is independent of chromosomes but dependent on spindle and amphiaster. The fate of DNA originally existing in the fused nucleus during achromosomal cleavage of fertilized sea urchin and starfish eggs induced by aphidicolin was determined using antidenatured DNA antibody. The nucleus is not formed in the divided daughter cells at the two-cell stage but the nuclear-envelope-free chromatin mass which is unassociated with mitotic apparatus remains in the center region of embryos, especially near the first cleavage furrow. These results indicate that the condensed and nonreplicated chromatin can not be associated with spindle and asters in the presence of aphidicolin.     

Ultrastructural study of developmental stages of Mattesia dispora (Neogregarinorida: Lipotrophidae), a parasite of the flour moth Ephestia kuehniella (Lepidoptera)

The ultrastructure of merozoites, gamonts and oocysts of the neogregarine Mattesia dispora and their development in larvae of the flour moth Ephestia kuehniella were studied by electron microscopy. The apical complex of free macronuclear merozoites was very distinct in micrographs of sections, the polar rings being especially prominent. Two gamonts associated in head-to-head syzygy and the apical complexes served as the contact point during pairing. At this stage the rhoptries became reduced and the conoid widened. The gamonts had a foam-like appearance in the light microscope. Paired gamonts formed an envelope and developed into a gametocyst, within which the gamonts were separated by a distinct border. Four gametes and two residual cells developed inside the gametocyst. The gametes were covered with a single membrane. The gametes fused in pairs to form two spherical zygotes, each covered by two membranes and with one large nucleus. The external layer appeared more undulated than the inner one. A single membrane covered each residual cell. Walls were formed around both zygotes to produce two oocysts. Each mature oocyst was lemon-shaped with polar plugs and eight peripheral sporozoites, which had a pellicle similar to that of the merozoites, lay beneath the thick oocyst wall.     

Mycamoeba gemmipara nov. gen., nov. sp., the First Cultured Member of the Environmental Dermamoebidae Clade LKM74 and its Unusual Life Cycle

Since the first environmental DNA surveys, entire groups of sequences called “environmental clades” did not have any cultured representative. LKM74 is an amoebozoan clade affiliated to Dermamoebidae, whose presence is pervasively reported in soil and freshwater. We obtained an isolate from soil that we assigned to LKM74 by molecular phylogeny, close related to freshwater clones. We described Mycamoeba gemmipara based on observations made with light‐ and transmission electron microscopy. It is an extremely small amoeba with typical lingulate shape. Unlike other Dermamoebidae, it lacked ornamentation on its cell membrane, and condensed chromatin formed characteristic patterns in the nucleus. M. gemmipara displayed a unique life cycle: trophozoites formed walled coccoid stages which grew through successive buddings and developed into branched structures holding cysts. These structures, measuring hundreds of micrometres, are built as the exclusive product of osmotrophic feeding. To demonstrate that M. gemmipara is a genuine soil inhabitant, we screened its presence in an environmental soil DNA diversity survey performed on an experimental setup where pig cadavers were left to decompose in soils to follow changes in eukaryotic communities. Mycamoeba gemmipara was present in all samples, although related reads were uncommon underneath the cadaver.     

Light optical and electron microscopy research on the cyst-forming coccidium Sarcocystis muris

Part of the complicated life cycle of Sarcocystis muris, confined to the muscle cyst (sarcocyst), has been studied by light and electron microscopy. The early development of the sarcocyst proceeds strictly intracellularly, whereas the older and larger cysts tend to destroy the harbouring muscle cell, and since then their development seems to be intercellular rather than intracellular. Three different cell types are distinguished within the growing sarcocyst of S. muris differing from each other both structurally and functionally: metrocytes, intermediate cells and merozoites. These differ as well in the structure of their nuclei. The metrocyte nuclear chromatin is mainly in decondensed state with some minute granules taking the central part of the nucleus. The condensed chromatin of the intermediate cell is accumulated into some relatively large peripheral granules, whereas numerous RNP-granules appear in the karyolymph. The nuclear chromatin of merozoites is condensed to be seen as separate chromocenters scattered over the nucleus; the karyolymph is packed with RNP-granules. Metrocytes are seen to divide in young sarcocysts, although the mode of their division is still obscure. In sarcocysts of advanced age (2.5 months or more), only intermediate cells are seen to divide, their mode of division being endodyogeny.     

中华血簇虫的生活史研究 Ⅰ.中华血簇虫在鳖穆蛭中的发育

中华血簇虫是湖北产的中华鳖体内发现的一种寄主虫。在其生活史发育中,存在着两种寄主的交替。本文只介绍它在无脊椎动物寄主——鳖穆蛭体中的发育情形。这一时期包括两个阶段:配子生殖和孢子生殖。配子生殖的特点是,两性配子母细胞先融合,然后才进行配子分化,产生4个雄配子核,其中1个核与雌配子核受精,形成合子核。孢子生殖以单核卵囊的核分裂开始,最后形成含8个裸子孢子的成熟卵囊,并解体释放出子孢子。整个过程都发生在蛭消化道内。脊椎动物寄主的感染,很可能是因为吃下含成熟子孢子的无脊椎动物寄主而引起的。     

Lankesteria barretti n. sp. (Eugregarinida,Diplocystidae), a Parasite of the Mosquito Aedes triseriatus (Say) and a Review of the Genus Lankesteria Mingazzini*

SYNOPSIS. Lankesteria barretti n. sp. (Eugregarinida, Diplocystidae) is named from the mosquito Aedes triseriatus in Texas. The young cephalins occur within the midgut epithelial cells. When they reach a length of about 150–200 μ they rupture the host cell and are released into the space between the epithelium and the peritrophic membrane, becoming gamonts. These grow to about 140–310 by 40 μ at the anterior end and 12 μ at the posterior end. When the host pupates they enter the lumen of the Malpighian tubules; pairs join in syzygy by their anterior ends and later more laterally. Each pair forms a spherical gametocyst about 60–100 μ (exceptionally 250 μ) in diameter. A large number of oocysts develop in each gametocyst. The mature oocysts are spindle-shaped, 11 by 5.4–5.7 μ, and contain 8 elongate sporozoites and a refractile residuum. The gametocyst wall breaks down, releasing oocysts in the Malpighian tubules of the host when it is adult. The oocysts pass out in the feces and presumably infect new larvae by ingestion. The cephalins and gamonts of L. barretti differ from those of L. culicis (of Aedes aegypti) in having a relatively anterior instead of a central nucleus and in lacking a noticeable mucron; its longitudinal folds are not as well-developed as in L. culicis, and its paraglycogen granules are larger. The fine structure of L. culicis and L. barretti is described in detail. Their gamonts have a polar ring but no definite conoid. The taxonomy of the genus is reviewed, but its species have been so poorly described that it is impossible to be sure whether they are all really Lankesteria. About 19 species have been described (5 from turbellaria, 8 from tunicates, perhaps 1 from Amphioxus, 1 from the chaetognath Sagitta sp., 1 from Phlebotomus and 3 from mosquitoes).     

The effect of progesterone on the localization of progesterone receptors in the nuclei of chick oviduct cells

Summary The location of occupied and unoccupied progesterone receptors (PR) in chick oviduct cells was studied by immuno-electron microscopy with the use of a highly specific polyclonal anti-PR antibody and pre-embedding modifications of the peroxidase-anti-peroxidase-(PAP-) or immunogold-silver methods. Both methods revealed a nuclear localization of the PRs. The location of the PR in the nucleus was studied in detail by means of the immunogold-silver method. The most intense labelling for unoccupied PRs was in the condensed chromatin. After occupation of PRs with progesterone (P), decondensation or dispersion of chromatin was observed. At the same time, the labelling in the border area of condensed and dispersed chromatin, and in the dispersed chromatin, increased. The changes were statistically significant. The results can be explained by conformational changes of the PR-containing chromatin rather than by translocation of PRs from one site to another.     

The site of action of the anticoccidial salinomycin (Coxistac).

The anticoccidial salinomycin has a cidal effect against chicken coccidia. Restricted and unrestricted medication studies and histopathological examinations of chicks infected with Eimeria acervulina, E. maxima, or E. tenella showed that parasites were destroyed within host cells during asexual development. Most sporozoites failed to become trophozoites and were destroyed 30--72 hr after ingestion of oocysts. The drug also affected schizonts during initial nuclear replication by either destroying or significantly delaying their maturation. Parasites affected by the drug were distorted grossly. Drug action against gametogony was not observed histologically, but when medication was restricted to this period of the life cycle, subsequent oocyst shedding of all 3 species was reduced by 20--70% compared to unmedicated controls. When drug was provided during the entire parasite life cycle, activity against asexual stages was so complete that only a limited number of parasites survived to form gamonts, and oocyst shedding was reduced by 80--90% relative to controls. As with other ionophores, salinomycin had no effect upon rate of oocyst sporulation.     

A cytological study of micronuclear elongation during conjugation in Tetrahymena

Micronuclear elongation is the first major event in a series of nuclear changes occurring during the sexual stage of the life cycle of Tetrahymena. Beginning at about one hour after cells of complementary mating types have conjugated, the micronucleus leaves its recess in the macronucleus and swells slightly. This is accompanied by a reorganization of its chromatin from a reticular to a solid body. In the next stage the micronucleus assumes an egg shape, a development concomitant with the appearance of microtubules. While the chromatin spins out from the dense body, and microtubules increase in number, the nucleus assumes a spindle shape. During the elongation, which increases the length of the nucleus some fifty fold, microtubules are prominent in clusters just internal to the nuclear membrane, and parallel to the longitudinal axis of the nucleus. When elongation is completed the nucleus is curved around the macronucleus. Internally, partially condensed strands of chromatin are located off-center, towards the macronuclear side, and the density of the microtubules is diminished. At all the stages, DNA is located throughout the nucleus; neither discrete chromosomes nor synaptonemal complexes are seen. Occasionally cytoplasmic membrane systems are seen fused to the nuclear envelope which retains the typical appearance of a double membrane with pores.     

Ultrastructure of the nuclear apparatus of the lower ciliateRemanella granulosa Kahl (Karyorelictida)

Summary The nuclear apparatus ofRemanella granulosa has been investigated using conventional TEM methods and Bernhard's technique of preferential RNP staining. This species has two (rarely three) macronuclei and a single micronucleus (rarely two micronuclei). The nuclei always form a single group.The macronuclei contain a fibro-granular matrix resistant to EDTA destaining, and several nucleoli and chromatin bodies. The chromatin bodies are readily bleached with EDTA and are often clustered, or even fused, forming chromocenters. The nuclei are of the compact concentric type. Some macronuclei contain nuclear bodies, as finely fibrous spheres or bundles of coarse fibers, or both. Neither type of nuclear body is destained with EDTA. The spheres are frequently associated with nucleoli. There is no evidence of any transition between the two types of nuclear bodies. The macronuclear envelope contains numerous pore complexes and is strengthened with an electron dense layer. The micronucleus is filled with spongy condensed chromatin and surrounded by an envelope with occasional pores. This nucleus lacks nucleoli and nuclear bodies.     

A mechanism of cohesin‐dependent loop extrusion organizes zygotic genome architecture

Fertilization triggers assembly of higher‐order chromatin structure from a condensed maternal and a naïve paternal genome to generate a totipotent embryo. Chromatin loops and domains have been detected in mouse zygotes by single‐nucleus Hi‐C (snHi‐C), but not bulk Hi‐C. It is therefore unclear when and how embryonic chromatin conformations are assembled. Here, we investigated whether a mechanism of cohesin‐dependent loop extrusion generates higher‐order chromatin structures within the one‐cell embryo. Using snHi‐C of mouse knockout embryos, we demonstrate that the zygotic genome folds into loops and domains that critically depend on Scc1‐cohesin and that are regulated in size and linear density by Wapl. Remarkably, we discovered distinct effects on maternal and paternal chromatin loop sizes, likely reflecting differences in loop extrusion dynamics and epigenetic reprogramming. Dynamic polymer models of chromosomes reproduce changes in snHi‐C, suggesting a mechanism where cohesin locally compacts chromatin by active loop extrusion, whose processivity is controlled by Wapl. Our simulations and experimental data provide evidence that cohesin‐dependent loop extrusion organizes mammalian genomes over multiple scales from the one‐cell embryo onward.