bluuebunny/crossref_metadata_2025 huggingface dataset:
ULTRASTRUCTURE OF DYADS IN MUSCLE FIBERS OF ASCARIS LUMBRICOIDES
Authors: Rosenbluth Jack
Published: May 2004

The dyads of Ascaris body muscle cells consist of flattened intracellular cisternae applied to the sarcolemma at the cell surface and along the length of T-tubules. In specimens prepared by conventional methods (glutaraldehyde fixation, osmium tetroxide postfixation, double staining of sections with uranyl acetate and lead hydroxide), both the sarcolemma and the limiting membrane of the cisterna exhibit unit membrane structure and the space between them is occupied by a layer of peg-shaped densities which is referred to as the subsarcolemmal lamina. The lumen of the cisterna contains a serrated layer of dense material referred to as the intracisternal lamina. In specimens fixed in glutaraldehyde, dehydrated, and then postfixed in phosphotungstic acid, with no exposure to osmium tetroxide or heavy metal stains, the membranous components of the dyads appear only as negative images, but the subsarcolemmal and intracisternal laminae still appear dense. Except for the lack of density in membranes and in glycogen deposits, the picture produced by the latter method is very much like that of tissue prepared by conventional methods.
INTERACTION OF ESTROGEN AND PROGESTERONE IN CHICK OVIDUCT DEVELOPMENT
Authors: Oka Takami , Schimke Robert T.
Published: May 2004

The effects of estrogen and progesterone on the function of chick oviduct tubular gland cells have been studied. Such function, as measured by the increase in specific cell products such as lysozyme and ovalbumin, requires the continuous presence of estrogen or progesterone. Withdrawal of hormone results in a rapid cessation of function and an involution of the oviduct accompanied by rapid decreases in total weight, lysozyme, and RNA. During such involution, tubular gland cells per se persist, as evidenced by a lack of comparable decrease in total DNA content and by histological demonstration of tubular gland cells. When estrogen administration is reinstituted, preexisting tubular gland cells rapidly synthesize ovalbumin and lysozyme without requiring new DNA synthesis. Administration of progesterone also stimulates the function of such cells. Furthermore, the effects of estrogen and progesterone are synergistic on the synthesis of lysozyme and ovalbumin, whereas progesterone antagonizes the estrogen-evoked formation of tubular gland cells. It is suggested that such complex interactions of estrogen and progesterone on oviduct development and function result from differences in responsiveness of the various cell types present in the tissue.
ELECTRON MICROSCOPIC RADIOAUTOGRAPHIC DETECTION OF SITES OF PROTEIN SYNTHESIS AND MIGRATION IN LIVER
Authors: Ashley Charles A. , Peters Theodore
Published: May 2004

The sites of synthesis of proteins and their subsequent migration in rat liver have been studied during a 75 min period after labeling of liver-slice proteins by exposure to leucine-H3 for 2 min. Incorporation of the label into protein began after 1 min and was maximal by 4 min. Electron microscopic radioautography showed that synthesis of proteins in hepatocytes occurs mainly on ribosomes, particularly those in rough endoplasmic reticulum and, to some extent, in nuclei and mitochondria. Most of the newly formed proteins leave the endoplasmic reticulum in the course of 40 min, and concurrently labeled proteins appear in Golgi bodies, smooth membranes, microbodies, and lysosomes. A likely pathway for the secretion of some or all plasma proteins is from typical rough endoplasmic reticulum to a zone of reticulum which is partially coated with ribosomes, to the Golgi apparatus, and thence to the cell periphery. The formation of protein by reticuloendothelial cells was measured and found to be about 5% of the total protein formed by the liver.
THE DEVELOPMENT OF GOLGI COMPLEXES AND THEIR DEPENDENCE UPON THE NUCLEUS IN AMEBAE
Authors: Flickinger Charles J.
Published: May 2004

The production of Golgi complexes was investigated in Amoeba proteus by introducing a nucleus into cells that had been enucleated for 5 days. Golgi complexes were not detected in 5 day enucleates, nor were they observed in amebae fixed 15 min after renucleation. Samples taken at longer intervals after the introduction of a nucleus exhibited an increase in the size and abundance of Golgi complexes. Small curved smooth cisternae, some of which were aligned in parallel to form small Golgi complexes, were observed 30 min after the operation. Aggregations of small Golgi complexes increased in number in amebae fixed 1 to 6 hr after renucleation. Golgi complexes of normal size were present 6 hr after the operation and became more abundant in samples fixed 12 hr, and 1, 2, and 3 days after renucleation. The possible participation of the granular endoplasmic reticulum in the development of Golgi complexes was suggested by two observations. First, the Golgi complexes in renucleates contained a dense material similar to the content of the endoplasmic reticulum in enucleates and early renucleates. Second, examples of continuity between the endoplasmic reticulum and Golgi cisternae were present in renucleates. The possibility that Golgi complexes can be produced in the absence of preexisting Golgi complexes is discussed.
INCREASE IN FUNCTION DURING GROWTH OF A SINGLE CELL ORGANELLE
Authors: Løvlie Arne
Published: May 2004

The gradient diver method has been adapted to measure photosynthesis during the early development of a single zygote of the multicellular green alga Ulva mutabilis Føyn. The cells have only one chloroplast, and the results indicate that during growth of the chloroplast the rate of O2-production increases linearly in the early part of the cell cycle but remains constant or decreases at the end of the cycle.
SPATIAL PATTERNS OF THREADLIKE ELEMENTS IN THE AXOPLASM OF THE GIANT NERVE FIBER OF THE SQUID (LOLIGO PEALII L.) AS DISCLOSED BY DIFFERENTIAL INTERFERENCE MICROSCOPY AND BY ELECTRON MICROSCOPY
Authors: Metuzals J. , Izzard C. S.
Published: May 2004

The giant nerve fiber of the squid (Loligo pealii L.) has been investigated in situ, and in fresh and fixed preparations, by differential interference microscopy and electron microscopy. A continuous, three-dimensional network, composed of threadlike elements, was disclosed in the axoplasm. The threadlike elements in the axoplasm are twisted as a whole into a steep, right-handed helix. In a peripheral ectoplasmic region, the elements are more parallel to one another and more densely packed than in a central endoplasmic core. The threadlike elements can be resolved into a hierarchy of decreasing order of size. Successive levels of the hierarchy are formed by the association of smaller elements into larger ones. The following levels in the hierarchy of network elements have been distinguished: 1–3-µ-wide threads, 0.1–0.35-µ-wide strands, and 70–250-A-wide unit-filament strands. The differential interference microscope selects, from the network, threads oriented at a specific angle to the long axis of the axon. The specific angle depends upon the orientation of the long axis of the axon relative to the direction of shear. It is postulated that the network configuration is expressed in the solid-state properties of the axoplasm essential for the normal functioning of the nerve fiber.
OSMOTICALLY-INDUCED ALTERATIONS IN VOLUME AND ULTRASTRUCTURE OF MITOCHONDRIA ISOLATED FROM RAT LIVER AND BOVINE HEART
Authors: Stoner Clinton D. , Sirak Howard D.
Published: May 2004

Detailed studies correlating changes in mitochondrial optical density, packed volume, and ultrastructure associated with osmotically-induced swelling were performed. Various swelling states were established by incubating mitochondria (isolated in 0.25 M sucrose) at 0°C for 5 min in series of KCl and sucrose solutions ranging in tonicity from 250 to 3 milliosmols. Reversibility of swelling was determined by examining mitochondria exposed to 250 milliosmols media after they had been induced to swell. Swelling induced by lowering the ambient tonicity to approximately 130 (liver mitochondria) and 90 (heart mitochondria) milliosmols involves primarily swelling of the inner compartment within the intact outer membrane. Decreasing the ambient tonicity beyond this level results in rupture of the outer membrane and expansion of the inner compartment through the break. The maximum extent of swelling, corresponding with complete unfolding of the cristae and an increase in over-all mitochondrial volume of approximately 6-fold (liver mitochondria) and 11-fold (heart mitochondria), is reached at approximately 15 (liver mitochondria) and 3 (heart mitochondria) milliosmols. Exposure of liver mitochondria to media of lower tonicity results in irreversibility of inner compartment swelling and escape of matrix material. These changes appear to result from increased inner membrane permeability, possibly due to stretching.
A SYSTEM OF PARALLEL SEPTA IN CRAYFISH NERVE FIBERS
Authors: Peracchia Camillo
Published: May 2004

Certain axons in the abdominal roots and nerve cord of crayfish contain a system of regularly spaced, parallel transverse septa with a periodicity of about 2 µ. Each septum is composed of two roughly parallel membranes, separated by a gap of 150–400 A. The two membranes are frequently fenestrated by pores 550–2000 A in diameter, each occupied by a microtubule. Filaments are occasionally seen bridging the gap between the microtubule and the edge of the pore. The membranes of the septa are continuous with longitudinal membranous tubules. In small- and medium-sized axons the septa are continuous across the axon, while in large axons they seem to be intact only at the periphery as annuli. It is suggested that such structures be called "fenestrated septa." With horseradish peroxidase as a tracer, no communication between the septal lumen and the periaxonal space was found.
BRUSH BORDER DEVELOPMENT IN THE INTESTINAL ABSORPTIVE CELLS OF XENOPUS DURING METAMORPHOSIS
Authors: Bonneville Mary A. , Weinstock Melvyn
Published: May 2004

The differentiation of the brush border which makes up the apical free surface of intestinal absorptive cells has been studied by electron microscopy. Specimens of Xenopus small intestine were fixed at various stages during metamorphosis, the time when a new intestinal epithelium forms. The interpretation of details described herein emphasizes the role of "surface-forming" vesicles. These vesicles are thought to provide membrane both for the initial expansion of the apical surface and for the later elongation of the microvilli. The latter are believed to be "molded" around filamentous cores that appear early in differentiation. The cores are attached to the apical membrane and extend vertically into the supranuclear cytoplasm. This interpretation rests chiefly on (a) the resemblance, both in morphology and in staining properties with colloidal thorium, between the membrane that limits the vesicles and that which limits the microvilli and (b) the distribution and time of appearance of the vesicles with respect to development of the microvilli. According to this view, the specific properties of surface membrane reside in preformed units that arise within the supranuclear cytoplasm. This morphogenetic process probably involves participation of the Golgi region as the site where the complex macromolecular architecture of the cell surface is assembled.
THE LOCALIZATION OF GLYCOGEN IN THE SPERMATOZOA OF VARIOUS INVERTEBRATE AND VERTEBRATE SPECIES
Authors: Anderson Winston A. , Personne Paul
Published: May 2004

With the periodic acid-thiosemicarbazide-silver proteinate procedure for the detection of polysaccharides in thin sections, glycogen is localized in the cavities of centrioles and basal bodies, within the axoneme (and surrounding it), in mitochondria, and in the "packing" cytoplasm of the middle piece of spermatozoa of several invertebrate and vertebrate species. The cytochemical localization of glycogen is verified by extraction with α-amylase solution. These findings establish the existence of stored glycogen in sperm. The polysaccharide presumably serves as an endogenous source of energy in the absence of extracellular metabolites, under either aerobic or anaerobic conditions. Other hypotheses on the physiological significance of intracellular glycogen stores in sperm are discussed. Sperm that store glycogen contain some enzymes of glycogen metabolism. In the presence of glucose-1-phosphate, ATP, and Mg++ ions, an amylophosphorylase catalyzes the in vivo synthesis of glycogen. The newly formed product resembles γ-particles, and is digestible with α-amylase.
MACROMOLECULAR PHYSIOLOGY OF PLASTIDS
Authors: Henningsen K. W. , Boynton J. E.
Published: May 2004

Sequential changes occurring in the etioplasts of the primary leaf of 7-day-old dark-grown barley seedlings upon continuous illumination with 20 lux have been investigated by electron microscopy, in vivo spectrophotometry, and thin-layer chromatography. Following photoconversion of the protochlorophyllide pigment to chlorophyllide and the structural transformation of the crystalline prolamellar bodies, the tubules of the prolamellar bodies are dispersed into the primary lamellar layers. As both chlorophyll a and b accumulate, extensive formation of grana takes place. After 4 hr of greening, protochlorophyllide starts to reaccumulate, and concomitantly both large and small crystalline prolamellar bodies are formed. This protochlorophyllide is rapidly photoconverted upon exposure of the leaves to high light intensity, which also effects a rapid reorganization of the recrystallized prolamellar bodies into primary lamellar layers.
FORMATION OF HORNY CELLS
Authors: Lavker Robert M. , Matoltsy A. Gedeon
Published: May 2004

Epithelial cells changing from the granular stage of differentiation to the horny stage are more numerous, and reveal sequential events of transformation in finer detail in the rumen epithelium than in other keratinizing epithelia thus far studied in the electron microscope. Studies of such cells indicate that transformation is initiated by the release of hydrolytic enzymes, as evidenced by the appearance of lysosomes. As lysosomes increase in number, the nucleus, ribosomes, mitochondria, Golgi apparatus, and mucous granules are gradually degraded. Furthermore, marked changes occur in permeability of the plasma membrane as voluminous amounts of the lysed cell components pass through and accumulate in the intercellular space in the form of an amorphous mass. Filaments, keratohyalin granules, and the content of the ER (ER-protein) are not lysed, revealing the action of released enzymes to be specific. During transformation, filaments become displaced toward the cell periphery and keratohyalin granules disperse and mix with the ER-protein in the cell center. Subsequently, the keratohyalin-ER-protein complex infiltrates the filament network whereby a fibrous-amorphous cell content is formed. Loss of fluids through the plasma membrane leads to reduction of cell volume and consolidation of the remaining cell content. The deep interdigitations formed between the cells ultimately interlock the outer part of the epithelium into a cohesive and protective stratum corneum.
CHLOROPLAST STRUCTURE AND FUNCTION IN ac-20, A MUTANT STRAIN OF CHLAMYDOMONAS REINHARDI
Authors: Goodenough Ursula W. , Levine R. P.
Published: May 2004

The fine structure of the ac-20 strain of Chlamydomonas reinhardi is described. Cells grown mixotrophically in the presence of acetate have a highly disordered chloroplast membrane organization and usually lack pyrenoids. Chloroplast ribosome levels are only 5–10% of wild-type levels. Cells grown phototrophically without acetate possess more chloroplast ribosomes and have more normal membrane and pyrenoid organization. Chloroplast ribosome levels rise rapidly when cells are transferred from acetate to minimal medium, whereas membrane reorganization occurs only after a lag. These results, combined with earlier studies of the photosynthetic properties of the mutant strain, suggest that proper membrane organization, Photosystem II activity, and ribulose-1,5-diphosphate carboxylase formation are dependent on the presence of chloroplast ribosomes. Other chloroplast components tested are unaffected by a 10-fold reduction in levels of chloroplast ribosomes.
BIOGENESIS OF CHLOROPLAST MEMBRANES
Authors: Goldberg I. , Ohad I.
Published: May 2004

The development of photosynthetic lamellae during greening of dark-grown Chlamydomonas y-1 cells was investigated by radioautography. Acetate-3H was used as a marker for membrane lipids. In short pulse-labeling experiments, about 50–60% of the radioactivity incorporated was found in the lipid fraction and about 25–50% in starch granules present in the chloroplast of these algae. The relative specificity of acetate-3H used as a marker for membranes was artificially increased through quantitative removal of the starch granules from fixed cells by amylase treatment. Analysis of turnover coefficients of different membrane constituents and of the contribution of turnover and net synthesis to the total label incorporated in pulse experiments indicated that the incorporation of acetate into specific lipids was mainly due to net synthesis. The distribution of radioactivity in the different lipid constituents at the end of a short pulse and after 30- and 60-min chases indicated that transacylation is minimal and may be disregarded as a possible cause of randomization of the label. Statistical analysis of radioautographic grain distribution and measurements of different structural parameters indicate that (a) the chloroplast volume and surface remain constant during the process, whereas the growth of the photosynthetic lamellae parallels the increase in chlorophyll; (b) the lamellae do not develop from the chloroplast envelope or from the tubular system of the pyrenoid; (c) all the lamellae grow by incorporation of new material within preexisting structures; (d) different types of lamellae grow at different rates. The pyrenoid tubular system develops faster than the thylakoids, and single thylakoids develop about twice as fast as those which are paired or fused to grana. It is concluded that growth of the membranes occurs by a mechanism of random intussusception of molecular complexes within different types of preexisting membranes.
SOME PROPERTIES OF HYALIN
Authors: Stephens R. E. , Kane R. E.
Published: May 2004

The principal protein component of the hyaline layer of sea urchin eggs is the calcium-insoluble protein first described by Kane and Hersh. The protein hyalin is abnormally high in acidic amino acids, almost devoid of basic amino acids, and characteristically rich in valine and proline. Essentially all of the cysteine present is found in the disulfide form; no evidence points to intermolecular disulfide linkages. Hyalin from several species has a minimal subunit weight of about 100,000, though evidence exists for a particle three times this weight in urea or guanidine hydrochloride from one species. Optical rotatory dispersion measurements indicate no α-helix content, though the dispersion has unique characteristic features. Addition of small quantities of calcium causes hyalin to gel to a birefringent fibrous form. The fibrous, birefringent form of hyalin is rendered isotropic upon addition of EDTA, but the birefringence is restored with re-addition of divalent cation.
CHANGES IN CHEMICAL COMPOSITION OF THYLAKOID MEMBRANES DURING GREENING OF THE y-1 MUTANT OF CHLAMYDOMONAS REINHARDI
Authors: De Petrocellis Benita , Siekevitz Philip , Palade George E.
Published: May 2004

Two fractions of thylakoid membranes (TMF) have been isolated from disrupted (French press) algal cells by using a discontinuous sucrose gradient. TMF-II consists mostly of thylakoid membranes still partially organized in grana; it contains also fragments of chloroplast envelope, pyrenoid tubules, and starch granules; thus it amounts to a fraction of chloroplast fragments which have lost practically all matrix components. TMF-I consists of smaller chloroplast fragments and is contaminated to a larger extent than TMF-II by other subcellular components, primarily mitochondria. TMF-II accounts for about 12% of the protein and 30% of the chlorophyll of the whole cell; it contains cytochrome 554 and carotenoids in the same ratio to chlorophyll as the latter, and shows photosystems I and II activities but lacks enzymatic activities characteristic of the dark reactions. During the greening of the y-1 mutant of Chlamydomonas, TMF's have been isolated over a range of chlorophyll concentrations from 5 to 25 µg/107 cells. The results showed that during this period the ratios of chlorophyll to cytochrome 554 and of chlorophyll to carotenoids, and the relative concentrations of individual carotenoids were continuously changing. The findings support the view that during greening, thylakoid membranes are produced by multistep assembly.
THE ISOLATION OF A MAJOR STRUCTURAL ELEMENT OF THE SEA URCHIN FERTILIZATION MEMBRANE
Authors: Bryan Joseph
Published: May 2004

Procedures for isolating the contents of the cortical granules from the ova of the sea urchin, Strongylocentrotus purpuratus, are reported. Dithiothreitol is used to remove the vitelline coat; the "demembranated" eggs are then subsequently activated with butyric acid. By means of these procedures, the hyaline protein and crystalline or paracrystalline material have been isolated from the cortical granules. The crystalline material consists of sheets of cylinders or tubules 150–200 A in diameter. This material is believed to be a major structural element of the fertilization membrane which, in the absence of the vitelline coat, does not form.
THE THREE-DIMENSIONAL RECONSTRUCTION OF THE XY CHROMOSOMAL PAIR IN HUMAN SPERMATOCYTES
Authors: Solari A. J. , Tres Laura L.
Published: May 2004

The spatial reconstruction of the XY pair of chromosomes from human spermatocytes has been made by the study of serial sections 1000 A in thickness. The sex pair during zygotene-pachytene forms a condensed mass of chromatin that has two filamentous, electron-opaque cores: the long and the short core. During early pachytene both cores have a common ending region, about 0.4–0.8 µ long. This common end is a synaptonemal complex, and each of the cores forms a lateral element of that complex. The cores become more convoluted during middle pachytene forming "ringlike bodies." Nucleoli from spermatocytes have three distinct regions: (a) granular; (b) dense fibrillar; and (c) clear intermediate. Occasional association of the XY pair and the heteropycnotic "basal knobs" results in apparent association of nucleoli and the sex pair in a minority of cells. The evidence presented is interpreted as a strong support of the hypothesis of homologous regions in the human XY pair.
CONTROLLED PRODUCTION OF PROLIFERATING SOMATIC CELL HYBRIDS
Authors: Klebe Robert J. , Chen Tchaw-Ren , Ruddle Frank H.
Published: May 2004

The techniques described permit the controlled production of large numbers of proliferating somatic cell hybrids in a relatively short period of time. Sendai virus is used to promote cell hybridization. ß-propriolactone is employed as the inactivating agent of Sendai virus since it produces complete loss of viral infectivity while preserving viral fusion capacity. Cells are fused in monolayer, instead of in suspension, since fixing cells in two dimensions permits one to control cell contacts during the fusion event through the expedient of varying multiplicities of the parental cells and the total cell density. Under the conditions described, a several hundred fold increase in the number of hybrid clones obtained is seen as compared to the controls.
ISOLATION, ULTRASTRUCTURE, AND BIOCHEMICAL CHARACTERIZATION OF GLYCOGEN IN INSECT FLIGHT MUSCLE
Authors: Childress Charles C. , Sacktor Bertram , Grossman I. William , Bueding Ernest
Published: May 2004

Glycogen from flight muscle of the blowfly, Phormia regina, has been characterized ultrastructurally and biochemically. In situ, glycogen is in the form of rosettes, which vary in size with diameters of up to 0.1 µ. Sedimentation analysis of pure glycogen, isolated by mild buffer extraction, reveals a polydisperse molecular weight spectrum, with larger particles having molecular weights of 100 million. Treatment of native glycogen with alkali, under conditions usual for the extraction of the polysaccharide from tissues, results in a 5- to 10-fold reduction in molecular weight, as well as a chemical alteration of the molecule. Flight muscle phosphorylase has a lower affinity for native than for alkali-treated glycogen. The maximum velocity of the enzyme is also lower with native substrate. The apparent Km for inorganic phosphate is higher with native glycogen as cosubstrate. These kinetic differences with native and partially degraded glycogen demonstrate the importance of using the natural substrate in studies of biochemical control mechanisms.
COMPARATIVE STUDIES ON MITOCHONDRIA ISOLATED FROM NEURON-ENRICHED AND GLIA-ENRICHED FRACTIONS OF RABBIT AND BEEF BRAIN
Authors: Hamberger Anders , Blomstrand Christian , Lehninger Albert L.
Published: May 2004

Fractions enriched in neuronal and glial cells were obtained from dispersions of whole beef brain and rabbit cerebral cortex by large-scale density gradient centrifugation procedures. The fractions were characterized by appropriate microscopic observation. Mitochondria were then isolated from these fractions by differential centrifugation of their homogenates. The two different types of mitochondria were characterized with respect to certain enzyme activities, respiratory rate, rate of protein synthesis, and their buoyant density in sucrose gradients. The mitochondria from the neuron-enriched fraction were distinguished by a higher rate of incorporation of amino acids into protein, higher cytochrome oxidase activity, and a higher buoyant density in sucrose density gradients. Mitochondria from the glia-enriched fraction showed relatively high monoamine oxidase and Na+- and K+-stimulated ATPase activities. The rates of oxidation of various substrates and the acceptor control ratios did not differ appreciably between the two types of mitochondria. The difference in the buoyant density of mitochondria isolated from the neuron-enriched and glia-enriched cell fractions was utilized in attempts to separate neuronal and glial mitochondria from the mixed mitochondria obtained from whole brain homogenates in shallow sucrose gradients. The appearance of two peaks of cytochrome oxidase, monoamine oxidase, and protein concentration in such gradients shows the potential feasibility of such an approach.
ELECTRON MICROSCOPY OF THE AMMONIACAL SILVER REACTION FOR HISTONES IN THE ERYTHROPOIETIC CELLS OF THE CHICK
Authors: MacRae Edith K. , Meetz Gerald D.
Published: May 2004

The product of the postformalin ammoniacal silver reaction, which has been claimed to distinguish lysine-rich from arginine-rich histones with the light microscope on the basis of a color difference, was examined in developing erythroid cells of chick bone marrow with the electron microscope. Stem cells and early erythroblasts exhibit no, or little, ammoniacal silver reaction product, while small basophilic erythroblasts, polychromatophilic erythrocytes, and reticulocytes exhibit an increasing amount of reaction product as maturation proceeds. The reaction product is in the form of discrete electron-opaque particles associated with heterochromatin. The ammoniacal silver reaction in the erythroid cell series is interpreted as reflecting either the accumulation of newly synthesized arginine-rich histones or changes in the availability of reactive sites in preformed histones.
SCALE FORMATION IN CHRYSOPHYCEAN ALGAE
Authors: Brown R. Malcolm , Franke Werner W. , Kleinig Hans , Falk Heinz , Sitte Peter
Published: May 2004

The cell wall of the marine chrysophycean alga Pleurochrysis scherfellii is composed of distinct wall fragments embedded in a gelatinous mass. The latter is a polysaccharide of pectic character which is rich in galactose and ribose. These wall fragments are identified as scales. They have been isolated and purified from the vegetative mother cell walls after zoospore formation. Their ultrastructure is described in an electron microscope study combining sectioning, freeze-etch, and negative staining techniques. The scales consist of a layer of concentrically arranged microfibrils (ribbons with cross-sections of 12 to 25 x 25 to 40 A) and underlying radial fibrils of similar dimensions. Such a network-plate is densely coated with particles which are assumed to be identical to the pectic component. The microfibrils are resistant to strong alkaline treatment and have been identified as cellulose by different methods, including sugar analysis after total hydrolysis, proton resonance spectroscopical examination (NMR spectroscopy) of the benzoylated product, and diverse histochemical tests. The formation and secretion of the scales can be followed along the maturing Golgi cisternae starting from a pronounced dilated "polymerization center" as a completely intracisternal process which ends in the exocytotic extrusion of the scales. The scales reveal the very same ultrastructure within the Golgi cisternae as they do in the cell wall. The present finding represents the first evidence on cellulose formation by the Golgi apparatus and is discussed in relation to a basic scheme for cellulose synthesis in plant cells in general.
INTERCELLULAR MIGRATION OF CENTRIOLES IN THE GERMARIUM OF DROSOPHILA MELANOGASTER
Authors: Mahowald Anthony P. , Strassheim Joan M.
Published: May 2004

A cluster of centrioles has been found in the early Drosophila oocyte. Since the oocyte is connected to 15 nurse cells by a system of intercellular bridges or ring canals, the possibility that the cluster of centrioles arose in the germarium from an intercellular migration of centrioles from the nurse cells to the oocyte was analyzed in serial sections for the electron microscope. Initially, all of the 16 cells of the future egg chambers possess centrioles, which are located in a juxtanuclear position. At the time the 16 cell cluster becomes arranged in a lens-shaped layer laterally across the germarium, the centrioles lose their juxtanuclear position and move towards the oocyte. By the time the 16 cell cluster of cells is surrounded by follicle cells (Stage 1), between 14 and 17 centrioles are found in the oocyte. Later, these centrioles become located between the oocyte nucleus and the follicle cell border and become aggregated into a cluster less than 1.5 µ in its largest dimension. The fate of these centrioles in the oocyte is not known. The fine structure of the germarium and the early oocyte is also described.
THE INTRACELLULAR LOCALIZATION OF INORGANIC CATIONS WITH POTASSIUM PYROANTIMONATE
Authors: Tandler Carlos J. , Libanati César M. , Sanchis Carlos A.
Published: May 2004

Potassium pyroantimonate, when used as fixative (saturated or half-saturated, without addition of any conventional fixative) has been demonstrated to produce intracellular precipitates of the insoluble salts of calcium, magnesium, and sodium and to preserve the general cell morphology. In both animal and plant tissues, the electron-opaque antimonate precipitates were found deposited in the nucleus—as well as within the nucleolus—and in the cytoplasm, largely at the site of the ribonucleoprotein particles; the condensed chromatin appeared relatively free of precipitates. The inorganic cations are probably in a loosely bound state since they are not retained by conventional fixatives. The implications of this inorganic cation distribution in the intact cell are discussed in connection with their anionic counterparts, i.e., complexing of cations by fixed anionic charges and the coexistence of a large pool of inorganic orthophosphate anions in the nucleus and nucleolus.
RNA IN CYTOPLASMIC AND NUCLEAR FRACTIONS OF CELLULAR SLIME MOLD AMEBAS
Authors: Cocucci S. M. , Sussman M.
Published: May 2004

A method is described for the rapid separation of cellular slime mold (Dictyostelium discoideum) cells into nuclear and cytoplasmic fractions. Sucrose density sedimentation profiles of radioactivity from cells that had been grown for long or short periods in the presence of uridine-3H indicate very low levels of cross-contamination between the fractions. The nuclear fraction contains few, if any, ribosomes. In exponentially growing cells, at least 80% of the ribosomes were associated in polysomal complexes. No loss of counts from pre-labeled rRNA was observed during 2 generations (24 hr) of logarithmic growth and, within the polysomal complexes, the distributions of the preformed material and of rRNA synthesized during the 2 generations were identical. In stationary phase cells that had entered the developmental program leading to fruiting body construction, the rRNA turned over rapidly so that by the end of development at least 75% of the ribosomes fabricated during exponential growth had disappeared and had been replaced by new ones synthesized during the morphogenetic sequence. The preformed ribosomes disappeared preferentially from the monosomal contingent; the newly synthesized ribosomes appeared exclusively in the polysomal contingent and did not appear as monosomes in appreciable numbers for at least 6 hr. The possible significance of this wholesale replacement of ribosomes is discussed.
THE FINE STRUCTURE AND ARRANGEMENT OF MICROCYLINDERS IN THE LUMINA OF FLAGELLAR FIBERS IN CRICKET SPERMATIDS
Authors: Kaye Jerome S.
Published: May 2004

Flagellar structure in spermatids of several species of cricket was studied with the electron microscope. The flagella of mid-spermatids contain the usual 9 + 2 set of fibers and a set of nine accessory fibers. At first all are hollow, then the lumina become filled with an electron-opaque matrix material in which narrow electron-lucent microcylinders are embedded. The accessory fibers and one central fiber become filled first, then the B subfibers and the other central fiber, and finally the A subfiber. In all but the B subfibers, microcylinders are arranged in a circular or oval group that lies against the wall of the lumen and encloses one or several additional microcylinders. In accessory fibers there are 9–11 microcylinders in the outer group and 4–5 in the inner group. In the central fibers and the A subfibers there are 7–9 microcylinders that enclose one or two more. In the B subfibers there is a crescentic group of 6–7 microcylinders that partially encloses 2–3 more. Microcylinders become packed as though they are independent units; the matrix appears to be an amorphous substance that fills the spaces around the microcylinders. The mean diameter of the microcylinders is 36 A, and they have a center-to-center distance of 56 A. In both respects they resemble wall subunits of flagellar fibers and microtubules and they may be similar structures but with a different localization. The diameter of accessory fibers is about 350 A, which is 25% greater than that of the other flagellar fibers and of cytoplasmic microtubules. Rotation tests suggest that the accessory fibers have 16 wall-subunits.
A CYTOCHEMICAL AND RADIOAUTOGRAPHIC STUDY OF HUMAN TISSUE CULTURE CELL NUCLEOLI
Authors: Recher L. , Whitescarver J. , Briggs L.
Published: May 2004

Fine structural aspects of human tissue culture cell nucleoli were studied by cytochemical and radioautographic methods. Ribonuclease and pepsin digestions were carried out on glutaraldehyde-fixed cells that, in some instances, were labeled with thymidine-3H prior to digestion. Double digestion by ribonuclease and pepsin revealed a fine fibrillar reticulum that appears to be the supportive structure of nucleolonemal threads. The nature of the reticulum remains to be determined. The question of whether it may represent a dispersed form of chromatin was raised. Structural findings suggested such an hypothesis but the results of radioautographic studies do not support it. The reticulum showed a striking absence of radioactive labeling following a 3 hr incorporation of thymidine-3H. Only few silver grains were observed occasionally in the fibrillar nucleolonema that may or may not be significant. The radioautographic results are believed to be inconclusive for the various reasons discussed. The possibility that the reticulum is composed of proteins has to be considered. It appears that basic proteins can resist pepsin digestion in aldehyde-fixed cells. Individual chromatin fibrils were found to be associated with the nucleolar reticulum. It is possible that these alone represent the dispersed genetically active chromatin of nucleoli.
MEMBRANE MODIFICATIONS IN NUTRITIONALLY INDUCED FILAMENTOUS ESCHERICHIA COLI B
Authors: Weinbaum George , Fischman Donald A. , Okuda Shinichi
Published: May 2004

Nutritionally induced filamentous cell forms of Escherichia coli B were examined for their morphological and biochemical lesions. The filamentous forms showed no significant alteration in total DNA concentration, RNA synthesis, ability to form ß-galactosidase in response to isopropylthiogalactoside, or insensitivity to actinomycin D as compared to the normal cell form. The filamentous cells showed a marked decrease in the ability to incorporate N-acetylglucosamine-UL-14C into a phenol-soluble glycoprotein fraction relative to the normal cell form or relative to strain E-26 of E. coli grown in the filament-inducing medium. The filaments yielded an envelope-specific phenol-soluble protein fraction markedly reduced in or lacking three proteins as determined by acrylamide gel electrophoresis. Amino acid analysis, and chemical and enzymatic treatments of the envelope-specific phenol-soluble proteins showed striking differences between the fractions obtained from normal and filamentous cells. Electron microscope studies of divalent cation-induced aggregates of the envelope proteins showed different aggregation patterns dependent upon the cell form yielding the protein fraction.
ABSENCE OF TRANSLATIONAL CONTROL OF HISTONE SYNTHESIS DURING THE HELA CELL LIFE CYCLE
Authors: Pederson Thoru , Robbins Elliott
Published: May 2004

The cell-free synthesis of histone-like polypeptides has been achieved using a selected class of small polyribosomes as the only particulate fraction. This synthesis is prevented if the deoxyribonucleic acid (DNA) inhibitor, cytosine arabinoside, is added to the cells prior to disruption, and it is not detected when the cytoplasm used is derived from postmitotic (G1) cells. When the 100,000 g supernate from pure metaphase populations was compared with that from S phase cells, the cell-free synthesis of histone-like polypeptides in the presence of S phase polyribosomes remained unchanged. These data suggest that, except for the histone messenger RNA-ribosome complex, the cytoplasmic factors requisite for histone synthesis are present throughout the cycle, and that the shut-off of this synthesis is not under translational control.
CILIA AND VESICULAR PARTICLES IN THE ENDOCRINE PANCREAS OF THE MONGOLIAN GERBIL
Authors: Boquist Lennart
Published: May 2004

Though the general appearance and the cellular composition of the pancreatic islets of the Mongolian gerbil (Meriones unguiculatus) conformed to those of most other rodent species, some peculiar ultrastructural details were found. Thus there were diversiform, mainly vesicular particles with varying electron opacity in these islets. The vesicular particles showed a clear association to cilia which seemed to possess a basic fiber pattern of 9 + 0. The basal bodies were localized in the cytoplasm of the islet parenchymal cells, most often in the ß-cells, and the vesicular particles occurred in the portions of cilia that were protruding into intercellular spaces. The cilia were often swollen, and the vesicular particles were mainly found in the space between the ciliary membranes and the longitudinal fibers. A few vesicular particles could be seen inside and sometimes seemingly in contact with these fibers. Occasionally, there were morphologically similar structures in the cytoplasm of adjacent ß-cells. The vesicular particles were differentiated from the vesicles occurring in nerve structures by their larger size, as well as by their heterogeneous shape and electron opacity. The nature of the vesicular particles and the significance of their presence in cilia and in the cytoplasm of some of the islet cells remain unknown. Among other possibilities, it is, however, suggested that the vesicular particles may represent secretory material.
ABERRANT INTRANUCLEOLAR MATURATION OF RIBOSOMAL PRECURSORS IN THE ABSENCE OF PROTEIN SYNTHESIS
Authors: Craig Nessly C. , Perry Robert P.
Published: May 2004

To help elucidate the role of protein in the maturation of ribosomal RNA in cultured L cells, we have studied the effects of cycloheximide upon the maturation process and upon the intranucleolar ribonucleoprotein particles containing the "preribosomal RNA's." Five parameters of these particles were analyzed: (a) extractability, (b) sedimentation characteristics in sucrose gradients, (c) RNA composition, (d) buoyant density in CsCl gradients, and (e) effects of increased ionic strength on the buoyant density. When protein synthesis is inhibited, the rate of conversion of the precursor 45S ribosomal RNA is rapidly diminished, falling to less than 30% of the control rate within 1 hr. Nevertheless, in terms of the first three parameters there is no difference between control and cycloheximide nucleolar particles. However, the cycloheximide particles have a lower and more heterogeneous buoyant density and a more variable response to increased ionic strength. The results imply that the protein composition of the cycloheximide particles is different from that of particles from control cells, and that the entire protein complement is not necessary for the first cleavages in the maturation process, although it is necessary for the normal rate of processing and for the eventual appearance of both 18S and 28S rRNA in mature ribosomes.
LOW RESISTANCE CONNECTIONS BETWEEN CELLS IN THE DEVELOPING ANTHER OF THE LILY
Authors: Spitzer Nicholas C.
Published: May 2004

Low resistance junctions were demonstrated between cells in anthers from young buds of Lilium longiflorum Croft by standard electrophysiological techniques. Electrodes containing a dye were used to stain impaled cells for later histological identification. Electrical coupling is widespread; germinal cells are coupled to one another; coupling is also observed between somatic elements, and germinal and somatic cells are similarly interconnected. Cytoplasmic bridges are implicated in the first case; plasmodesmata are probably responsible for the interactions in the other two. Although the physiological role of the low resistance junctions shown here and present in embryonic animal tissues is unknown, the possible function of this form of intercellular communication in the development of the anther is discussed.
RIBOSOME SYNTHESIS IN TETRAHYMENA PYRIFORMIS
Authors: Kumar Ajit
Published: May 2004

The cellular site of synthesis of ribosomal RNA in Tetrahymena pyriformis was studied by analyzing the purified nuclear and cytoplasmic RNA from cells pulse labeled with uridine-3H. The results of studies using zonal centrifugation in sucrose density gradients show that the ribosomal RNA is synthesized in the nucleus as a large precursor molecule sedimenting at 35S. The 35S molecule undergoes rapid transformation through two main nuclear intermediates, sedimenting at about 30S and 26S. The smaller ribosomal RNA (17S) appears first in the cytoplasm and it seems to be absent from the nucleus. The apparent delay in the appearance of the larger ribosomal RNA (26S) in the cytoplasm is due to the presence of a larger pool of its precursors in the nucleus as indicated by pulse-chase experiments. The newly synthesized ribosomal RNA's appear in the cytoplasm as discrete 60S and 45S ribonucleoprotein particles, before their incorporation into the polysomes.
THE NATURE AND PROCESSING OF RIBOSOMAL RIBONUCLEIC ACID IN A DINOFLAGELLATE
Authors: Rae Peter M. M.
Published: May 2004

Certain features of the dinoflagellate nucleus suggest that it represents a primitive form of eukaryotic nucleus. For this reason, it was of interest to characterize dinoflagellate ribosomal RNA (rRNA) and its mode of synthesis to determine if it also deviated from typical eukaryotic patterns. Gyrodinium cohnii was chosen for this examination. Gyrodinium ribosomal RNA species are 16 and 25s as judged by their sedimentation velocities in isokinetic sucrose gradients. These values are typical of higher plants. In addition, the RNA cosedimented precisely with rRNA from the ciliate Tetrahymena. Nucleotide ratio analyses revealed a GMP + CMP content of 46% for both species of rRNA. The kinetics of incorporation of a radioactive precursor into ribosomal RNA have also been studied, and it seems likely that the maturation of rRNA starts with the synthesis of a 38s molecule. This serves as precursor to the 16s species, and, after a 27s intermediate, the 25s ribosomal component. The process is similar to that in other eukaryotes. The structure of the nucleolus has also been examined, and is seen to be typically eukaryotic.
RESPIRATION AND PROTEIN SYNTHESIS IN ESCHERICHIA COLI MEMBRANE-ENVELOPE FRAGMENTS
Authors: Hendler Richard W. , Nanninga N.
Published: May 2004

The membranous nature of pellets obtained from broken Escherichia coli spheroplasts by successive centrifugation at 3500 g (P1), 20,000 g (P2), and 105,000 g (P3), has been established by electron microscopy. Spectrophotometric analysis has shown that about 90% of the cytochromes are concentrated in the particulate fractions. The crude ribosomal pellet (P3) contained as much of the total cytochromes as did the pellet obtained at 20,000 g (P2). The high cytochrome content of P3 is consistent with its high oxidative activity (1) and the presence of membrane vesicles in this fraction. Analysis at 77°K intensified the optical extinction of all the cytochrome absorption bands, but the degree of intensification was not uniform for each fraction nor for each band within a given fraction. Carbon monoxide had little or no inhibiting effect on NADH oxidation. Reduced plus carbon monoxide difference spectra yielded artifactual absorption bands in the wave length regions where reduced vs. oxidized absorption bands normally occur. Succinate and NADH, either together or separately, reduced nearly all of the cytochromes, indicating that the cytochrome portion of the electron-transport chain is shared by both substrates. A tentative formulation of the electron-transport chain is presented.
THE SURFACE CHARGE OF RAT LIVER MITOCHONDRIA AND THEIR MEMBRANES
Authors: Heidrich Hans-G. , Stahn Roland , Hannig Kurt
Published: May 2004

The surface charge of intact mitochondria and submitochondrial particles was examined by the technique of preparative free flow electrophoresis. When submitochondrial preparations obtained by a swelling-contraction procedure were examined with this technique, two fractions were observed. One of these fractions exhibited the same electrophoretic properties as intact mitochondria, which indicated that it was derived from the outer limiting membrane of the mitochondrion. This fraction was found to contain the enzymes monoamine oxidase and rotenone-insensitive NADH-cytochrome c reductase which have been reported to be localized in the outer mitochondrial membrane. The other fraction exhibited an electrophoretic mobility which was different from that of intact mitochondria, and this fraction contained enzymes characteristic of the inner membrane-matrix fraction such as soluble and particulate enzymes of the Krebs cycle. Microsomes exhibited an electrophoretic mobility which was almost identical with that of the outer mitochondrial membrane. In addition to resolving the localization of enzymes in mitochondrial membranes, these data indicate that the outer limiting membrane of the mitochondrion is the sole determinant of the surface charge of mitochondria.