Identification and DNA sequence analysis of a γ-type gliadin cDNA plasmid from winter wheat

Summary Recombinant cDNA plasmids possessing the coding sequences for the -type gliadins were isolated from a cDNA library prepared from wheat seed poly (A⁺) RNA. One of these plasmids, pGliB48, specifically hybridizes to poly (A⁺) RNA molecules 1 400–1 500 bases in length that direct the synthesis of polypeptides at 38 Kd and 46 Kd, the latter size characteristic of the -type gliadins. The cDNA sequence of pGliB48 was determined and encompasses the 3 untranslated region as well as 245 amino acids from the C-terminus of the -type gliadin polypeptide. The 5-end of the DNA coding sequence consists of a tandem repeat unit composed of eight amino acids. Localized regions of homology are observed for the /-type and -type gliadin cDNA sequences.     

Structure of root nodules formed by Rhizobium on the non-legume Trema cannabina var. scabra

The structure of the nodules formed by Rhizobium on the non-legume Trema cannabina var. scabra was studied using the light microscope. The overall features of the nodules showed greater resemblance to the non-legume rather than the legume nodule. Nodule squashes yielded bundles of infection threads and bacteroids with morphological differences from rhizobial cells grown on yeast-mannitol-glucose agar. Two types of cell infection occurred within the bacterial zone; plant cells were either, like legumes, filled with rhizobia released from the infection threads (less than one third of infected cells) or were filled with the extensive growth and development of the infection thread. The rate of nitrogen fixation in the Trema nodule was high. It seemed that host cells filled with threads were active in N fixation.     

Predators and avian community organization: an experiment in a semi-desert grassland

Summary We provide experimental evidence that predators are a major factor organizing a community of granivorous grassland birds (mostly emberizid finches). Our focus is not on the lethal effects of predators, but on the simple idea that (i) birds will not settle where they perceive a high risk of predation, and (ii) species differ in their perception of the safety of woody vegetative cover due to differences in antipredator escape behavior. Consistent with this idea is the fact that the composition of this grassland community responds markedly to minor manipulations in the distribution of woody cover. In particular, with the addition of cover to open grasslands, species with cover-dependent escape tactics increase in abundance, while the abundance of cover-independent species decreases greatly; this decrease may reflect aggression from cover-dependent species, but evidence suggests that some cover-independent species may actively avoid cover-rich areas per se. Non-predatory effects of cover, most notably those concerning food resources and microclimate, appear unable to explain these results. Predators may influence many communities of terrestrial vertebrates via species-specific responses to cover.     

Effects of light on nitrate-limited Oscillatoria agardhii in chemostat cultures

The effects of the average light irradiance (I) on growth and nitrate uptake kinetics of the cyanobacterium Oscillatoria agardhii, in nitrate-limited chemostat cultures, were studied. Light was nonsaturating for I <9.4 Wm^–2, for all growth rates () studied. However, was throughout limited by the availability of nitrate. Under light-saturating conditions the kinetics of nitrate-limited growth could be adequately described by both the Monod and Droop equations. Under light-non-saturating conditions the internal nitrogen content (Q) was a function of both and I, for which new formulas were derived. The high uptake capacity (V _max) of nitrate-limited cells was independent of , but was significantly increased for cells growing at I <9.4 Wm^–2. The half-saturation constant for nitrate uptake (K _s ^u ) increased with increasing , but was independent of the prevailing light conditions. The effects of light during nitrate-limited growth were associated with the regulation in the nitrogen-containing pigments.The results reported herein have important consequences for the use of Q, K _s ^u and V _max values as indicators of nutrient-deficiency of natural populations.     

Electron microscopic studies on the adenohypophysis of the White-Crowned sparrow,Zonotrichia leucophrys gambelii

Summary The pars distalis of the adenohypophysis of normal (78), thyroideotomized (6), adrenalectomized (6), and castrated (14) White-crowned Sparrows were observed with the electron microscope. Six types of glandular cells were identified and the ultrastructural characteristics of each have been described. To each has been assigned tentatively an endocrine function.STH cells are characterized by the presence of large, dense secretory granules ranging from 220–280 m, a poorly developed endoplasmic reticulum, and a fragmented Golgi apparatus; they occur only in the caudal lobe. They show no remarkable changes after adrenalectomy, castration, and thyroidectomy.Prolactin cells, whose identity is suggested by their responses to photostimulation and surgical experiments, are characterized by large, polymorphic, dense secretory granules; they have been found mainly in the cephalic lobe.ACTH cells, whose function is confirmed by their cytological responses to adrenalectomy, have a peculiar type of secretory granule (220 m) with high and low phases of electron density. They occur exclusively in the cephalic lobe and are transformed, after adrenalectomy to large, vacuolated adrenalectomy cells.TSH cells are so designated by their response to thyroidectomy. After thyroidectomy, they lose their specific fine secretory granules and are transformed into large, vacuolated thyroidectomy cells. We have found TSH cells and thyroidectomy cells only in the cephalic lobe.Two types considered to be gonadotropic cells from their responses to gonadectomy, occur in both the cephalic and caudal lobes. One of them contains spherical, dense secretory granules (180–220 m), prominent rough endoplasmic reticulum and a well developed Golgi apparatus; the other type contains dense secretory granules of variable size (150–350 m), a less extensively developed Golgi apparatus, and sac-like endoplasmic reticulum. Both types of gonadotropic cells show extreme enlargement and vacuolization after castration. However, they retain differences in appearance in the structure of cytoplasmic organelles and vacuolization.Dedicated to Professor Dr. H. Grau in honor of his 70th birthday.The investigation reported herein was supported by a research grant (HE 07240 NEUA) from the National Institutes of Health to Professor Vitums, by a research grant (5R01 NB 06187) from the National Institutes of Health to Professor Farner, and by a scientific research grant (No. 91049) from the Ministry of Education of Japan to Professor Mikami. The authors wish to thank Professor James R. King for his assistance in obtaining and maintaining the birds, and for his helpful advice concerning the experiments.     

Three-dimensional organization of fibroblasts and collagen fibrils in rat tail tendon

Summary The orderly arrangement of fibroblasts and collagen in tendons and ligaments suggests that these cells may have precise relationships with one another and with the collagen fibrils. The spatial organization of rat tail tendon was therefore examined using scanning and transmission electron microscopy and by reconstructing a 35-m long segment of tendon from serial transmission electron micrographs. Fibroblasts were regularly arranged in columns and showed more intimate association in the longitudinal than in the transverse plane. Thin cytoplasmic sheets extended up to 3 m transversely, frequently forming junctional attachments with similar processes from adjacent cells or from the same cell. Longitudinal processes were longer, often extending for more than 20 m and forming junctional attachments with other cells in the same column. Such processes often exhibited invaginations in which there were single fibrils or small groups of fibrils; this arrangement may be indicative of fibril elongation or may serve to transmit tension between the fibroblast and the collagen fibrils. This organization has interesting implications for the growth and function of other fibrous connective tissue, such as the periodontal ligament.     

On the histology and ultrastructure of the Harderian gland in rabbits

Summary The Harderian gland of rabbits has been studied with light and electron microscopes. The red part contains relatively wide alveoli with an irregular cuboidal epithelium. The stainability of the cytoplasm is poor. The white part has smaller alveoli with a low columnar epithelium. The cells show cytoplasmic basophilia removable with ribonuclease. The cytoplasm of both kinds of cells is very dense when examined in the electron microscope. The mitochondria show branched, closely packed cristae and a dense matrix. The Golgi apparatus displays few lamellae and rows of vacuoles. The endoplasmic reticulum is very finemeshed and partly associated with ribonucleoprotein particles. Both kinds of cells contain numerous lipid droplets, leaving vacuoles in the sections prepared for electron microscopy. They are fewer but distinctly larger in the red part. In both lobes pictures suggesting a secretion of lipid droplets have been observed. Cells showing signs of degeneration with subsequent discharge of the detritus have been observed in both lobes, but this process does not correspond to the holocrine secretion in sebaceous glands. Likewise, no apocrine secretion was observed.     

Microbodies inCrithidia fasciculata

Summary Electron microscopy of thin sections ofCrithidia fasciculata showed cytoplasmic organelles 0.2–0.8 m in diameter containing a finely granular matrix limited by a single membrane. Utilization of cytochemical techniques (DAB) for endogenous catalase revealed pronounced deposition of a highly electron-opaque material in these organelles. Reaction of cells with DAB was completely inhibited by 0.02 M 3-amino-1,2,4-triazole. It is concluded that this organelle is a microbody on the basis of cytological and cytochemical similarities with microbodies from plant and other animal cells.This work was supported in part by research grant NIAID-USPHS AI09445.     

Reactions of seven basic fluorochromes with unfixed cells obtained from the salivary glands of the dipteran fly Megaselia scalaris Loew (Phoridae)

Summary Seven basic fluorochromes with varying specificities were used to stain the large squamous epithelial cells isolated from the larval salivary glands of Megaselia scalaris (Phoridae). Although the EDTA-based method selected for isolating the cells produced permeabilization and a loss of viability of the cells, consistent results were obtained with the various fluorochromes. The classical pattern of green nuclear and red cytoplasmic fluorescence observed in cells stained with acridine orange could be changed to green cytoplasmic and red nuclear fluorescence by pretreatment with RNase. The predominantly cytoplasmic and nucleolar fluorescence obtained with pyronine Y could be changed to mainly nuclear fluorescence by RNase pretreatment. The other five fluorochromes tested were not affected appreciably by extraction with RNase. Quinacrine mustard, dicarbocyanine (DiOC₆(3)), and rhodamine 123 produced primarily cytoplasmic and nucleolar fluorescence, while nile red revealed mainly cytoplasmic lipid droplets. Phosphine 3R initially stained lipid droplets but very rapidly redistributed throughout the cytoplasm and nucleus. Because of their large size, flatness, and content of histochemically demonstrable components, the cells of Megaselia are especially appropriate for use as optical objects or controls in various studies. New methods of isolating the cells, however, will be needed to prevent permeabilization and loss of viability of the cells.     

Quantitative topography of organelles in the liver

Summary After seven days of feeding fructose the liver of Wistar rats showed enormous accumulations of glycogen, which completely altered the original pattern of distribution of organelles. A quantitative morphological method was used to analyze these changes.The cytoplasm was mapped into arbitrary distance classes corresponding to concentric rings beginning at the outer nuclear membrane. This allowed the density of organelles in a given zone to be estimated.In cells filled with glycogen as a result of the fructose feeding, the following rearrangements were found: in the intermediate zone of both cellular poles (i.e., bile canalicular pole and sinusoidal pole) the mitochondria disappeared, being replaced by glycogen.The endoplasmic reticulum was accumulated in the perinuclear zone of both cellular poles, as in control animals, but was reduced throughout the rest of cytoplasm. It showed a peripheral density maximum at the biliary canalicular pole, in contrast to the cells of control animals.These changes in the distribution of the organelles and cellular compartments correspond to histochemical findings and demonstrate an adaptive reaction in the liver parenchyma to fructose ingestion, the organelles arranging themselves in cytoplasmic regions which still show a metabolic activity.Supported by a grant from the Deutsche Forschungsgemeinschaft Az Ri 271/6-5     

Membranes and organelles of dehydratedSelaginella andTortula retain their normal configuration and structural integrity

Summary Dry (7–10% water content) leaves of the spikemossSelaginella lepidophylla (resurrection plant) and of the desiccationtolerant moss,Tortula ruralis were examined by freeze fracture electron microscopy. As has been described for dry seeds, the cells of these dehydrated leaves were shrunken, with highly convoluted walls and membranes. The membranes of all samples had a lipid bilayer organization with dispersed intramembranous particles (IMPs). Lipid droplets were very closely associated with the plasmamembrane. Chloroplasts were surrounded by a double membrane envelope and contained well-organized grana. Mitochondria were irregular in outline, and endoplasmic reticulum and cytoplasmic vesicles were present.Abbreviations ABA abscisic acid - EF exoplasmic fracture - FTIR Fourier transform infrared analysis - H_II hexagonal II - IMPs intramembranous particles - MGDG monogalactosyl diacylglycerol - NMR nuclear magnetic resonance - PE phosphatidylethanolamine - PF protoplasmic fracture - PS I photosystem I - PS II photosystem II     

Lipids in the proximal convoluted tubule of the cat kidney and the reabsorption of cholesterol

Summary Lipid deposits in the cat kidney are mainly located in the epithelium of the proximal tubuli contorti, particularly in the pars contorta. As the amount of fatty acids in the blood of renal arteries is higher than in renal veins, the lipid inclusions are likely to be formed in the proximal convoluted tubule. Whether fat occurring in the urine has been released from the nephron epithelium and the mode of this release remains obscure. The structural equivalent of lipid extrusion into the tubules has not been observed.Components of the tubular lipids include triglycerides, phosphoglycerides and cholesterol. The results of the digitonin-cholesterol reaction favour the assumption that cholesterol is eliminated in the glomeruli and pinocytotically reabsorbed by the brush border cells, this process possibly serving recycling of this compound. The dilated basal labyrinth and intercellular space contain perpendicularly oriented lipid accumulations that reach the basal lamina. The ultrastructure of the lipid storing cells of pars contorta reacting positively for phosphoglyceride and cholesterol is characterised mainly by bodies with marginal plates. As far as can be judged from their morphology, these bodies are interpreted as large peroxisomes. A special feature of the pars recta are dumbbell shaped bodies and elongated or cup-like mitochondria concentrically surrounding cytoplasmic areas, as well as a well-developed smooth ER. In what way the organelles of the brush border cells are involved in catabolic and anabolic processes as far as renal lipid metabolism is concerned remains to be answered.This investigation was supported by a grant from the Deutsche ForschungsgemeinschaftThis paper is dedicated in friendship to Professor Berta Scharrer (New York) on the occasion of her 70th birthday     

Stage-specific localization of cytoskeletal actin mRNA in murine seminiferous tubules and intestinal epithelia as demonstrated by in-situ hybridization

Summary In-situ hybridization experiments have been performed using isoactin ( and )-specific riboprobes in various tissues of the rat and mouse. Distribution of the grains of actin mRNAs for both and types was similar throughout sections of the rat testis. Although both mRNAs were evenly distributed in the seminiferous tubule, extremely heavy labeling was observed in about 10% of the seminiferous tubules that could be identified as stage XII of spermatogenesis. At high magnification, grains of the mRNA were found in the cytoplasm of elongating spermatids and in the Sertoli cell cytoplasm at the adluminal side. Much higher density of the grains of mRNA was observed in the neck region of the spermatids at stage XII. Thus, the dense distribution of cytoskeletal actin mRNAs is stage-specific in the tubule during spermatogenesis in the rat. The high expression of both and actin mRNAs was also observed in the epithelial cells of the intestinal crypts.     

Transport and Retention Mechanisms Govern Lipid Droplet Inheritance in Saccharomyces cerevisiae

Lipid droplets are ubiquitous cellular structures involved in energy homeostasis and metabolism that have long been considered as simple inert deposits of lipid. Here, we show that lipid droplets are bona fide organelles that are actively partitioned between mother cell and daughter cell in Saccharomyces cerevisiae. Video microscopy revealed that a subset of lipid droplets moves from mother cell to bud in an ordered, vectorial process, while the remaining lipid droplets are retained by the mother cell. Bud‐directed movement of lipid droplets is mediated by the molecular motor Myo2p, while retention of lipid droplets occurs at the perinuclear endoplasmic reticulum. Lipid droplets are thus apportioned between mother cell and daughter cell at cell division rather than being made anew.      

Ultrastructural studies on the vitellogenesis of Tetrodontophora bielanensis (Waga) (Collembola)

Summary Vitellogenesis in Tetrodontophora bielanensis (Waga) is of the mixed type. Part of the yolk material is produced inside the oocyte (auto-synthesis), while part is absorbed by micropinocytosis. During autosynthesis polyribosomes, rough endoplasmic reticulum and dictyosomes take part. Regardless of their origin, mature yolk spheres are constructed identically and are composed of three elements: cortex layer, matrix and crystals. Histochemical tests show that polysaccharides are present in the yolk spheres. Lipid droplets have been observed in the ooplasm; they develop without visible contact with any of the organelles. Among the reserve materials the following have been found: rough endoplasmic reticulum, dictyosomes, polyribosomes, mitochondria and a few microtubules.     

Presence of abnormally high incidences of sister chromatid exchanges in three successive cell cycles in Bloom's syndrome lymphocytes

The frequencies of sister chromatid exchanges (SCEs) were examined in phytohaemagglutinin-stimulated blood lymphocytes of a normal individual, a Bloom's syndrome heterozygote (bl/+), and two Bloom's syndrome homozygotes (bl/bl). To determine the baseline SCE frequencies, lymphocytes were cultured with various concentrations of 5-bromodeoxyuridine (BrdUrd) for two cell cycles. The incidence of SCEs per two cell cycles inbl/bl lymphocytes levelled off at BrdUrd concentrations below 10 g/ml while that in normal andbl/+ lymphocytes stayed constant below 7.5 g/ml. The baseline SCE frequency in bl/bl cells was ten times higher than that in normal andbl/+ cells. At BrdUrd concentrations above 15 g/ml, SCEs inbl/bl cells were induced more frequently than in normal andbl/+ cells. These results indicate that at low concentrations BrdUrd has a minimal effect on the induction of SCEs in all individuals, while at higher concentrations the BrdUrd incorporated inbl/bl cells has a larger effect than that in normal andbl/+ cells. To elucidate the effect of BrdUrd incorporated into the daughter and parental DNA strands on SCE induction, SCEs occurring during each cell cycle were examined separately in three-way or two-way differentially stained, third-cycle metaphases. The incidence of SCEs detected in each cell cycle at 5 g/ml BrdUrd was constant in all individuals and the rates of SCEs in each cell cycle inbl/bl cells were remarkably higher than those observed in normal andbl/+ cells. These findings strongly indicate that most of the abnormally increased SCEs in thebl/bl cells used in our study occurred independently of any effect of BrdUrd incorporated into both the daughter and parental DNA strands. In addition, an abnormal response ofbl/bl cells to BrdUrd was not found for cell cycle progression or chromosomal aberration induction. Thus, the bl/bl cells did not exhibit an abnormal hypersensitivity to BrdUrd. From these results, it seems quite probable that the abnormally increased SCEs in thebl/bl lymphocytes used here were spontaneous.     

Ultrastructure of elastic cartilage in the rat external ear

Summary The structure of elastic cartilage in the external ear of the rat was investigated by transmission and scanning electron microscopy.The narrow subperichondrial, boundary zone contains predominantly ovoid cells rich in cell organelles: mitochondria, Golgi complex, granular endoplasmic reticulum and small (40–100 nm) vesicles. Scarce glycogen granules and bundles of 6–7 nm cytoplasmic filaments are also present. Deeper in the boundary zone, one or more cytoplasmic lipid droplets appear and cytofilaments become more abundant.Fully differentiated chondrocytes in the central zone of the cartilage plate resemble white adipose cells. They are globular and contain a single, large cytoplasmic lipid droplet. The cytoplasm is reduced to a thin peripheral rim; it contains a flattened nucleus, few cytoplasmic organelles and abundant, densely packed, cytoplasmic filaments.The intercellular matrix is very sparse. The pericellular ring consists of collagen fibrils about 20 nm in diameter and a proteoglycan cartilage matrix in the form of a stellate reticulum. The complex of these two structures appears in the scanning electron micrographs as a network of randomly oriented, ca 100 nm thick fibrils. Spaces between pericellular rings of matrix also contain thick elastic fibers or plates, apparently devoid of microfibrils. In scanning electron micrographs elastic fibers could be detected only in a few areas, in which they were not obscured by other constituents of the matrix. Immature forms of elastic fibers, oxytalan (pre-elastic) and elaunin fibers, were found in the perichondrial and boundary zones.     

Fine-structural characteristics of the liver of the cod (Gadus morhua macrocephalus), with special regard to the concept of a hepatoskeletal system formed by Ito cells

Summary The liver of the adult cod (teleost, Gadus morhua macrocephalus) was observed with transmission and scanning electron microscopy. Hepatocytes of this animal are extremely large (about 50–70 m in diameter) and characterized by numerous large lipid droplets (5–15 m in diameter) showing fluorescence for vitamin A, though weaker than that of Ito cells. No Kupffer cells were recognized in the endothelial lining. Collagen fibrils are sparse in the perisinusoidal space, while Ito cells stretching their long cytoplasmic processes to the perisinusoidal as well as interparenchymatous spaces are frequent. There are a number of large desmosomes between the cytoplasmic processes, and all Ito cells seem to be interconnected by these junctions. The cytoplasm in the cell bodies and cell processes is occupied by bundles of intermediate filaments, while organelles are poorly developed. Small vesicles and caveolae are arranged along the plasma membrane. Scanning electron microscopy shows distinct three-dimensional networks consisting of Ito cells and their processes, which might be supporting elements of the liver tissue. We wish to emphasize the concept of this hepatoskeletal system.     

Day-night differences in the number of pineal “synaptic” ribbons in two diurnal rodents,the chipmunk (Tamias striatus) and the ground squirrel (Spermophilus richardsonii)

Summary Daytime numbers of pineal synaptic ribbons higher than reported in the pineal gland of any other mammalian species were observed in two diurnal rodents, the eastern chipmunk and Richardson's ground squirrel. The number of synaptic ribbons was lower during the daytime and higher at night in both of these species.     

The macronuclear envelope ofTetrahymena pyriformis GL in different physiological states

Summary A simple formula is derived to calculate the nucleocytoplasmic RNA-Efflux per nuclear Pore complex per min (REP-rate) which is generally applicable both for growing and stationary eukaryotic cells. In actively growing cells this REP-rate is mainly dependent on the cytoplasmic RNA-pool, the number of RNA-transporting pores, and the growth constant of RNA. These parameters are determined in logarithmicTetrahymena pyriformis GL. In this organism, 45 molecules both of the larger ribosomal RNA (25s rRNA) and of the smaller (17s rRNA) are transported per pore per min from nucleus to cytoplasm. Pulse-label experiments with³H-uridine indicate that the 25s rRNA is obviously transferred more slowly to the cytoplasm than the 17s rRNA. We postulate a gating hypothesis on the regulation of the nucleocytoplasmic RNA-efflux by nuclear pore complexes. This gating hypothesis suggests that nucleopores are controlling points of secondary importance in the sequence of gene expression, and do not directly control the cytoplasmic protein synthesis in eukaryotic cells.