Light and electron microscopy on the sporulation of the oocysts of Eimeria brunetti. I. Development of the zygote and formation of the sporoblasts

The initial stages of sporulation in oocysts of Eimeria brunetti were examined in samples sporulated at 27 degrees C for 0, 12 and 24 hours. The initial zygote was found to be roughly spherical and to contain a number of polysaccharide granules which were congregated in one region of the organism. The cytoplasm also contained some strands of rough endoplasmic reticulum together with a number of mitochondria, some Golgi bodies, and some electron translucent vacuoles. The nucleus was large, with amorphous nucleoplasm and a nucleous. The cytoplasmic mass of the zygote was limited by a single unit membrane which possessed some micropores. After initiation of the sporulation, the metabolic activity of the organism appeared to increase as evidenced by the augmentation in the cytoplasm of the amounts of rough endoplasmic reticulum, number of Golgi bodies, and the appearance of polyribosomes. However, at this stage, the presence of large spherical bodies (anlagen of the refractile bodies of the sporozoites) constituted the most obvious change in the cytoplasm of the organism. After nuclear division the daughter nuclei were situated well separated in the cytoplasm and the polysaccharide granules were evenly distributed throughout the cytoplasm of the zygote. Eventually four sporoblasts were formed by invaginations of the limiting membrane. Each sporoblast was limited by a unit membrane and contained a nucleus, and the same cytoplasmic organelles as found in the zygote. The development of the sporoblast was initially accompanied by the appearance of a second limiting membrane.     

Light and electron microscopy on the sporulation of the oocysts of Eimeria brunetti. II. Development into the sporocyst and formation of the sporozoite

The later stages of sporulation in oocysts of Eimeria brunetti were examined in samples which had been allowed to sporulate at 27 degrees C for 24, 36 and 48 hours. It was observed that the sporoblasts became ellipsoidal and the nucleus underwent the final division. A nucleus with associated Golgi bodies was not observed at either end of the organism. The cytoplasm was limited by two unit membranes and contained rough endoplasmic reticulum, dense bodies, electron translucent vacuoles and mitochondria. The first evidence of sporozoite formation was the appearance of a dense plaque at either end of the organism. This appeared in the vicinity of the nuclei, and adjacent to the limiting membrane of the soroblast. At this stage the sporocyst wall was still unformed. Then the two sporozoites were formed from opposite ends of the organism by growth of the dense plaques and invaginations of the plasmalemma which thus formed the pellicles of the developing sporozoites. A conoid and subpellicular microtubules were observed at this stage as development continued, a number of vacuoles were found between the nucleus and the conoid. These vacuoles constituted the precursors of the rhoptries and micronemes. At the same stage a large dense body had appeared within the forming sporozoite. As the sporozoite developed, this body, anterior refractile body, is followed by the nucleus and another dense body which formed the posterior refractile body. During this period, the thin sporocyst wall was formed and Stieda and sub-Stieda bodies were now present at one end of the sporocyst. Each mature sporocyst contained two sporozoites.     

Ultrastructural observations on fertilization and sporulation in Goussia iroquoina (Molnar and Fernando, 1974) in experimentally infected fathead minnows (Pimephales promelas, Cyprinidae)

The ultrastructural features of fertilization and sporogony of Eimeria iroquoina are described from the intestinal epithelium of experimentally infected fathead minnows (Pimephales promelas). Intact microgametes were observed in the cytoplasm of macrogametes. Within immature macrogametes the microgamete was segregated from the cytoplasm of the former by the plasma membrane of each cell plus additional membranes. Within mature macrogametes, only the plasma membranes separated the gametes. Fertilization by fusion of the limiting membrane of both gametes occurred after the entire microgamete lay within the cytoplasm of the macrogamete. The cytoplasm of the zygote cleaved into sporoblasts within cisternae of endoplasmic reticulum. The sporocystic wall was composed of an outer electron-lucent layer and an inner, thicker layer with periodic striations at right angles to the surface of the sporocyst. The sporocysts were bivalved and joined by a continuous suture. The sporozoites were morphologically similar to sporozoites and merozoites of other Coccidia. Due to the structure of the sporocyst, Eimeria iroquoina Molnar and Fernando, 1974 is amended to Goussia iroquoina (Molnar and Fernando, 1974).     

The effect of dehydration on the ultrastructure and cholinesterase activity of the subcommissural organ in the rat

Summary Dehydration affected certain cytological features of the subcommissural organ in the albino rat suggesting a strong secretory stimulation of the ependymal and hypendymal cells of this organ in dehydrated animals.The cytoplasm of the secretory cells of the subcommissural organ in the dehydrated rats was filled with dilated and empty sacs and vacuoles of endoplasmic reticulum. The membrane system of the Golgi apparatus was also dilated, and more numerous vesicles and vacuoles of the Golgi complex were noticed after dehydration.In brains of the dehydrated animals, Reissner's fibre was not found in the lumen of the third ventricle, and only a few vesicles containing homogeneous secretory material were seen in the cytoplasm of the subcommissural secretory cells.In control animals, the activities of the specific and non-specific cholinesterases were localized in the cytoplasmic and nuclear membranes as well as in the rough and smooth endoplasmic reticulum. After dehydration, the activities of the specific and non-specific cholinesterases were strongly decreased.     

OBSERVATIONS ON THE MODE OF RELEASE OF HERPES VIRUS FROM INFECTED HELA CELLS

The development of a well adapted strain of herpes virus has been studied in HeLa cells using thin sectioning techniques for electron microscopy. Particular attention was directed to events in the cytoplasm and certain new features were observed. Profuse immature particles with a nucleoid and single limiting membrane were present in the nuclei of infected cells, often in crystalline array; morphologically indistinguishable immature particles were also found very frequently in the cytoplasm. Cells with such particles were intact and well preserved, and contained smooth vacuoles apparently derived from the Golgi component of the endoplasmic reticulum. The cytoplasmic particles escaped from the cells by bulging out as buds through the cell membrane or through that of the cytoplasmic vacuoles until they were attached only by a pedicle and then became free. During this process the particles were gradually enclosed by the membrane through which they passed and carried a coat of it with them as they matured. After permanganate fixation the triple-layered structure of the cell membrane and vacuolar membranes was evident and was identical with that of the outer coat of the mature virus. These findings are discussed both in relation to different types of virus structure and to function in the endoplasmic reticulum and cell membrane.     

Structural changes in the granulosa lutein cells of pregnant cows between 60 and 245 days.

The structural changes in the granulosa lutein cells in the pregnant cows between 60 and 245 days, were observed. The polyhedral or pump cells has an acidophilic cytoplasm around the spherical nucleus. The glycogen granules increased in number up to 230 days of pregnancy. The lipid globules increased in size and amount during advanced pregnancy. The mitochondria showed a wide range of variations in shape and size. The matrix of many mitochondria contained dense inclusions which replaced the entire matrix in late pregnancy. The endoplasmic reticulum was chiefly of smooth type but a few stacks of the rough variety could be seen in the early days of pregnancy. Sometimes, whorls of smooth-surfaced endoplasmic reticulum enclosing mitochondria were seen. The Golgi complex was fully developed in the granulosa cells of all cases studied. The dense granules and lipid globules increased in size and number in the older corpora lutea. The cell surface showed numerous infolding of the plasma membrane.     

The mechanism of cytoplasmic streaming in characean algal cells: sliding of endoplasmic reticulum along actin filaments

Electron microscopy of directly frozen giant cells of characean algae shows a continuous, tridimensional network of anastomosing tubes and cisternae of rough endoplasmic reticulum which pervade the streaming region of their cytoplasm. Portions of this endoplasmic reticulum contact the parallel bundles of actin filaments at the interface with the stationary cortical cytoplasm. Mitochondria, glycosomes, and other small cytoplasmic organelles enmeshed in the endoplasmic reticulum network display Brownian motion while streaming. The binding and sliding of endoplasmic reticulum membranes along actin cables can also be directly visualized after the cytoplasm of these cells is dissociated in a buffer containing ATP. The shear forces produced at the interface with the dissociated actin cables move large aggregates of endoplasmic reticulum and other organelles. The combination of fast-freezing electron microscopy and video microscopy of living cells and dissociated cytoplasm demonstrates that the cytoplasmic streaming depends on endoplasmic reticulum membranes sliding along the stationary actin cables. Thus, the continuous network of endoplasmic reticulum provides a means of exerting motive forces on cytoplasm deep inside the cell distant from the cortical actin cables where the motive force is generated.     

Structure of taste buds in foliate papillae of the rhesus monkey, Macaca mulatta

Taste buds in foliate papillae of the rhesus monkey were examined by electron microscopy. Three distinct cell types were identified. Type I cells were narrow elongated cells containing an oval nucleus, bundles of intermediate filaments, several Golgi bodies, and characteristic apical membrane-bounded dense granules. These cells exhibited morphological variations: some had a moderately dense cytoplasm, perinuclear free ribosomes, and flattened sacs of rough endoplasmic reticulum; others had a more lucent cytoplasm, dilated irregular rough endoplasmic reticulum, lysosome-like dense bodies, and lipid droplets. Type II cells typically contained a spherical, pale nucleus, a prominent nucleolus, supranuclear and infranuclear Golgi bodies, mitochondria with tubular cristae, and one or two centrioles. This cell type, too, showed some variation in the relative amounts of ribosomes and smooth endoplasmic reticulum, which varied inversely with each other. Type III cells were characterized by a clear apical cytoplasm essentially devoid of ribosomes and containing microtubules. In a few type III cells, the peri- and infranuclear regions contained many ribosomes and some rough endoplasmic reticulum. In most Type III cells, there were large numbers of dense and clear vesicles in the peri- and infranuclear regions; some of the vesicles were grouped in synapse-like arrangements with adjacent nerves. The morphological variations exhibited by all three cell types could be accounted for by age differences in each of the cells. This would be consistent with the notion that cell renewal occurs in each of the three cell populations.     

Role of the nuclear envelope in synthesis, processing, and transport of membrane glycoproteins

The outer nuclear membrane is morphologically similar to rough endoplasmic reticulum. The presence of ribosomes bound to its cytoplasmic surface suggests that it could be a site of synthesis of membrane glycoproteins. We have examined the biogenesis of the vesicular stomatitis virus G protein in the nuclear envelope as a model for the biogenesis of membrane glycoproteins. G protein was present in nuclear membranes of infected Friend erythroleukemia cells immediately following synthesis and was transported out of nuclear membranes to cytoplasmic membranes with a time course similar to transport from rough endoplasmic reticulum (t 1/2 = 5-7 min). Temperature-sensitive mutations in viral membrane proteins which block transport of G protein from endoplasmic reticulum also blocked transport of G protein from the nuclear envelope. Friend erythroleukemia cells and NIH 3T3 cells differed in the fraction of newly synthesized G protein found in nuclear membranes, apparently reflecting the relative amount of nuclear membrane compared to endoplasmic reticulum available for glycoprotein synthesis. Nuclear membranes from erythroleukemia cells appeared to have the enzymatic activities necessary for cleavage of the signal sequence and core glycosylation of newly synthesized G protein. Signal peptidase activity was detected by the ability of detergent-solubilized membranes of isolated nuclei to correctly remove the signal sequence of human preplacental lactogen. RNA isolated from the nuclear envelope was highly enriched for G protein mRNA, suggesting that G protein was synthesized on the outer nuclear membrane rather than redistributing to nuclear membranes from endoplasmic reticulum before or during cell fractionation. These results suggest a mechanism for incorporation of membrane glycoproteins into the nuclear envelope and suggest that in some cell types the nuclear envelope is a major source of newly synthesized membrane glycoproteins.     

Studies on vinblastine-induced autophagocytosis in mouse liver. IV. Origin of membranes

The origin of the limiting membranes of autophagic vacuoles (AV) in mouse hepatocytes was studied by cytochemical techniques. Autophagocytosis was induced by an intraperitoneal injection of vinblastine (50 mg/kg). The marker enzymes used were adenosine triphosphatase for the plasma membrane, glucose-6-phosphatase for the endoplasmic reticulum and thiamine pyrophosphatase for the Golgi apparatus and the endoplasmic reticulum. All the three enzymes showed a characteristic localization in both control and vinblastine-treated hepatocytes. The space between the limiting membranes of a few apparently newly formed AV's showed weak glucose-6-phosphatase activity. Neither adenosine triphosphatase nor thiamine pyrophosphatase activities were observed on or between the AV membranes. It was suggested that endoplasmic reticulum membranes may be used as a source of AV membranes in hepatocytes. The lack of glucose-6-phosphatase activity in the limiting membranes even of most of the newly formed AV's suggests a transformation process of the membranes destined to form AV, during which the enzyme activity characteristic for endoplasmic reticulum may disappear from them.     

甜菊愈伤组织分生区细胞中膜内含物的超微结构研究

生长在分化培养基上的甜菊（Ｓｔｅｖｉａ ｒｅｂａｕｄｉａｎａ）愈伤组织分生区细胞中存在双膜和多膜内含物。电镜观察表明,这些膜内含物是由一圈或多圈呈同民贺或卷绕状排列的内质网包围部分细胞质而形成的。双膜内含物内外层膜的靠细胞质表面有核糖体附着,而多膜内含物仅在其最外层潴泡的外膜上偶有和量核糖体附着。附着细胞液泡化程度的提高,多膜内含物通过液泡膜内陷而转移到液泡中或通过消化其中被包围的细胞质及内膜而转     

Biogenesis of endoplasmic reticulum membrane in rat liver cells. II. Discharge of the nascent peptides of NADPH-cytochrome c reductase and cytochrome b5 on the cytoplasmic side of the endoplasmic reticulum membrane.

The direction of discharge of the nascent peptides of NADPH-cytochrome c reductase and cytochrome b5 from bound polyribosomes of rough microsomes was investigated in order to elucidate the mechanism of separation of these membrane proteins from secretory proteins, which are also synthesized by the same class of ribosomes of rough endoplasmic reticulum. The nascent peptides of NADPH-cytochrome c reductase and cytochrome b5 in intact rough microsomes were accessible to externally added 125I-Fab's against these proteins, and were susceptible to trypsin digestion, whereas the nascent peptides of serum albumin were not. The nascent peptides of these two microsomal proteins were released into the cytoplasm by puromycin treatment of intact rough microsomes, while the nascent peptides of serum albumin were retained in the microsomal lumen. These observations suggest that the nascent peptides of microsomal proteins, which are present on the cytoplasmic surface of the endoplasmic reticulum membrane, are exposed on the surface of microsomal vesicles, while those of secretory proteins are enclosed inside the vesicles. Therefore, the topographical separation of microsomal membrane proteins from secretory proteins is accomplished at the step of their synthesis by the bound polyribosomes of rough endoplasmic reticulum.     

Identification and subcellular distribution of adipocyte peptides and phosphopeptides.

Subcellular fractions of high purity (including plasma membrane, endoplasmic reticulum, mitochondria, nuclei, and cytoplasm) were prepared from isolated adipocytes, and the peptide components were examined by detergent gel electrophoresis. Each fraction except the endoplasmic reticulum exhibited a unique and reproducible complement of major peptides. Although the endoplasmic reticulum was distinctive in its enzymic markers, its peptide components showed striking homologies with certain species in the plasma membrane and cytoplasm. The two major adipocyte glycopeptides appear to be contained in the plasma membrane, inasmuch as they followed the distribution of 5'-nucleotidase. Incubation of adipocytes with extracellular 32Pi led to a uniform rate of incorporation of 32P into cellular peptides, with steady-state incorporation reached by 2 hours. Plasma membrane, mitochondria, nuclei, and cytoplasm all contained a distinctive complement of from two to five major phosphopeptides of different molecular weights. The majority of endoplasmic reticulum phosphopeptides exhibited molecular weights closely similar to those of certain species in the plasma membrane and cytoplasm. The phosphopeptides of the plasma membrane exhibited the highest absolute 32P incorporation of all phosphopeptides, next was the single major mitochondiral phosphopeptide. All fractions except the mitochondria contained, in addition to the few major phosphopeptides, numerous minor 32P-labeled phosphopeptides.     

Ultrastructural studies on the sporulation of oocysts of Toxoplasma gondii. II. Formation of the sporocyst and structure of the sporocyst wall

The ultrastructural changes observed during sporocyst formation and the structure of the sporocyst wall was examined in oocysts which had been allowed to sporulate for between 12 and 48 hours at 27 degrees C. As the spherical sporoblast developed into the sporocyst the cytoplasmic mass became ellipsoidal in shape although no change was noted in the organelle compliment, which cosisted of two nuclei plus a number of polysaccharide granules, lipid globules, mitochondria, Golgi bodies, and some rough endoplasmic reticulum. The sporocyst wall consisted of a thin outer layer (15-20 nm) which was formed from two limiting membranes of the sporoblast and an inner layer (40-50 nm) which was comprised of four curved plates. This inner layer was formed under the outer layer and, although no specific cytoplasmic organelle disappeared with its formation, some unit membranes were observed close to the plasmalemma during its formation. Each curved plate has a marginal swelling and an interposing strip of material is present between the margins of adjacent plates. The plates are joined to the interposing strip by a thin band of osmiophilic material. In oblique and tangential sections through the plates two types of cross banding were observed which differed in periodicity.     

The ultrastructure of macrogametes of Eimeria ferrisi Levine and Ivens 1965 in Mus musculus.

Macrogametes of Eimeria ferrisi occurred in epithelial cells of the cecum and colon of Mus musculus and were studied by electron microscopy. Young stages were identified as macrogamonts by the presence of wall-forming bodies. At first an outerlimiting membrane and remnants of the inner membrane complex of the former merozoite pellicle were present; the latter was later lost but in mature macrogametes 3 limiting membranes were observed. Type II wall-forming bodies appeared before type I; the former developed in expanded cisternae of the endoplasmic reticulum whereas the latter were smaller in size and appeared in the ground substance of the cytoplasm. After formation of the oocyst wall the bodies of the 2 types were no longer visible. The presenceodies of the 2 types were no longer visible. The persistence of micronemes in mature macrogametes and the presence of numerous layers of rough endoplasmic reticulum during wall formation have not been previously reported.     

CELL DIVISION IN SPIROGYRA. I. MITOSIS

Dividing cells of Spirogyra sp. were examined with both the light and electron microscopes. By preprophase many of the typical transverse wall micro-tubules disappeared while others were seen in the thickened cytoplasmic strands. Microtubules appeared in the polar cytoplasm at prophase and by prometaphase they penetrated the nucleus. They were attached to chromosomes at metaphase and early anaphase, and formed a sheath surrounding the spindle during anaphase; they were seen in the interzonal strands and cytoplasmic strands at telophase. The interphase nucleolus, containing 2 distinct zones and chromatinlike material, fragmented at prophase; at metaphase and anaphase nucleolar material coated the chromosomes, obscuring them by late anaphase. The chromosomes condensed in the nucleoplasm at prophase, moving into the nucleolus at prometaphase. The nuclear envelope was finally disrupted at anaphase during spindle elongation; at telophase membrane profiles coated the reforming nuclei. During anaphase and early telophase the interzonal region contained vacuoles, a few micro-tubules, and sometimes eliminated n ucleolar material; most small organelles, including swollen endoplasmic reticulum and tubular membranes, were concentrated in the polar cytoplasm. Quantitative and qualitative cytological observations strongly suggest movement of intact wall rnicrotubules to the spindle at preprophase and then back again at telophase.     

Studies on the endoplasmic reticulum. II. Simple dispositions in cells in situ

A survey of a large number of different cell types has indicated the presence of a network of membrane-bound cavities (the endoplasmic reticulum) in the cytoplasm of all cell types examined, with the exception of the mature erythrocyte. In its simplest form, encountered in seminal epithelia and in leucocytes, the reticulum consists mainly of interconnected strings of vesicles and appears to be randomly disposed in three dimensions. Local differentiations occur within the endoplasmic reticulum of all the cell types studied. The membrane limiting the cavities of the endoplasmic reticulum appears to be continuous with the cell membrane and the nuclear membranes.     

THE INTRACELLULAR MEMBRANES OF BLASTOMYCES DERMATITIDIS

Edwards , George A., and Mercedes R. Edwards . (Div. Labs, and Research, N.Y.S. Dept. of Health, Albany.) The intracellular membranes of Blastomyces dermatitidis. Amer. Jour. Bot. 47 (8): 622–632. Illus. 1960.—The yeast cells of Blastomyces dermatitidis have been studied in thin sections with the electron microscope. The cell is multinucleate, and the nuclei are frequently interconnected by their outer limiting membranes. The cell is bordered by a cell wall and the plasma membrane, which may be seen in direct continuity with the nuclear envelope. The cytoplasm contains numerous mitochondria, many profiles of the endoplasmic reticulum, and few multivesicular bodies. The membranes of all the constant cellular components are interconnected. Mitochondria appear to be formed from any of several membrane systems. The micromorphology of the cell suggests efficiency of communication and cytoplasmic mobility.     

Cytoplasmic cisterns structurally linked with the plasmalemma in the hepatocytes of mice of different lines]

Structures of the granular endoplasmic reticulum and narrow canals formed by smooth membranes were found in the cell membrane of male mice belonging to strains CBA, C57BL and L-NZB/BLN. The canals were seen connected with plasma membranes. Under the conditions of CC1(4) poisoning of CBA and C57BL mice, these canals were transformed to gigantic cytoplasmic vacuoles. Their cavities were communicated with the extracellular space. It still remains unclear if the membranes of these canals belong to the cytoplasmic membrane or to the endoplasmic reticulum.     

A tubular configuration of the granular endoplasmic reticulum forming a raft-like parallel array in the pinealocytes of two species of Japanese moles (Mogera kobeae and M. wogura)

Summary Ten or more straight tubules, each of which consists of a double unit membrane of granular endoplasmic reticulum with a cylindrical profile, are joined side by side in a raft-like configuration in the cytoplasm of the pinealocytes of Japanese moles. They measure about 60 nm and 100 nm in their inner and outer diameters, respectively, and are often partially connected to unspecialized granular endoplasmic reticulum. Cisterns held between the inner and outer unit membranes with cylindrical profiles vary from 15 nm to 30 nm in width. Ensheathed portions of the cytoplasm are contiguous with cytoplasm outside the tubular units. The inner unit membranes of the tubules bear fewer ribosomal particles than the outer ones.