Ultrastructural changes in the endoplasmic reticulum during dormancy release of apple embryos (Pyrus malus L.)

Apple embryos were treated by cold (0°C) within the fruits, to break their dormancy; the controls were treated at 12°C or at 20°C. Ultrastructural features of meristematic cells in the embryonic axis were compared for each treatment. The organization of the cells of dormant embryos was described: Endoplasmic reticulum consisted in some short rough cisternae; lipid droplets regularly arranged near the plasmalemma constituted a kind of shell; mitochondria had a few cristae; and dictyosomes were rarely observed. All these features are typical of dry seeds. After cold treatments, the only evolution observed was in the endoplasmic reticulum, where highly organized stacks appeared progressively as a function of time at 0°C. An intermediate temperature (12°C) induced similar formations in the reticulum but they were rarely observed and their degree of organization was lower than that obtained at 0°C. At 20°C, endoplasmic reticulum resembled that of the dormant embryo cells. The relation between the appearance of these structures in the reticulum and the disappearance of dormancy induced by cold is discussed.Abbreviations ER endoplasmic reticulum - RER rough endoplasmic reticulum     

Associations between membranes and microtubules during mitosis and cytokinesis in caulonema tip cells of the mossFunaria hygrometrica

Summary The interphase nucleus in theFunaria caulonema tip cells is associated with many non-cortical microtubules (Mts). In prophase, the cortical Mts disappear in the nuclear region; in contrast to moss leaflets, a preprophase band of Mts is not formed in the caulonema. The Mts of the early spindle are associated with the fragments of the nuclear envelope. Remnants of the nucleolus remain in the form of granular bodies till interphase. The metaphase chromosomes have distinct kinetochores; the kinetochore Mts are intermingled with non-kinetochore Mts running closely along the chromatin. Each kinetochore is associated with an ER cisterna. ER cisternae also accompany the spindle fibers in metaphase and anaphase. In telophase, Golgi vesicles accumulate in the periphery of the developing cell plate where no Mts are found. The reorientation of the cell plate into an oblique position can be inhibited by colchicine. It is concluded that the ER participates in controlling the Mt system, perhaps via calcium ions (membrane-bound calcium ions have been visualized by staining with chlorotetracycline) but that, on the other hand, the Mt system also influences the distribution of the ER. The occurrence and function of the preprophase band of Mts is discussed.     

Effect of supraoptimal temperatures on the function of the subcortical fibrils and an endoplasmic factor inNitella internodes

Summary The effect of supraoptimal temperatures onNitella cells was studied with special reference to the function of subcortical fibrils and an endoplasmic factor. Local heat-treatment (50 °C for 1 minute) of an internodal cell ofNitella disclosed that 1. the subcortical fibrils in the treated area remain normal, not affected by the treatment, 2. the subcortical fibrils alone produce no cytoplasmic streaming, 3. the endoplasm contains an extremely heat-labile factor which is indispensable for streaming, and 4. the stagnant endoplasm in the heat-treated area is neither coagulated nor gelated by heat.Preliminary reports appeared in Proc. 37th Annu. Meet. Bot. Soc. Jpn. P. 160 (1972, in Japanese) and in Abst. Annu. Meet. Jpn. Soc. Cell Biol. p. 57 (1975, in Japanese).     

Differentiation of Ca2+ pumps linked to plasma membrane and endoplasmic reticulum in the microsomal fraction from intestinal smooth muscle

ATP promotes ⁴⁵Ca uptake by the microsomal fraction from the longitudinal smooth muscle of guinea-pig ileum and this uptake is stimulated by oxalate. As the microsomal fraction is made up of various subcellular entities, we examined the localization of the Ca²⁺-transport activity by density gradient centrifugation, taking advantage of the selective effect of digitonin (at low concentration) on the density of plasmalemmal elements. When the ⁴⁵Ca-uptake activity was measured in the absence of oxalate, its behavior in subfractionation experiments closely paralleled that of the plasmalemmal marker 5′-nucleotidase. In contrast, the additional Ca²⁺-transport activity elicited by oxalate behaved like NADH-cytochrome $\text{c}$ reductase, a putative endoplasmic reticulum marker. The endoplasmic reticulum vesicles constituted only a small part of the membranes in the microsomal fraction, which explains that their Ca²⁺-storage capacity was not detectable in the absence of Ca²⁺-trapping agent. Low digitonin concentrations selectively increased the Ca²⁺ permeability of the plasmalemmal vesicles. The two Ca²⁺-transport activities were further differentiated by their distinct sensitivities to K⁺, vanadate and calmodulin. In this respect, the oxalte-insensitive and oxalate-stimulated Ca²⁺-transport systems resembled, respectively, the sarcolemmal and sarcoplasmic reticulum Ca²⁺ pumps in cardiac and skeletal muscle, in accordance with the subcellular locations established by density gradient centrifugation.     

Ultrastructural reactions of spinal ganglia to tri-ortho-cresyl-phosphate: Effects of neurotoxicity

Summary The ultrastructure of neurons in spinal ganglia of the domestic fowl poisoned with tri-ortho-cresyl-phosphate (TOCP) shows characteristic changes. The light neurons react to TOCP by a marked increase in the number of neurofilaments. These neurons also contain mitochondria in various degenerative stages. Several of the altered mitochondria show an increasing osmiophilia. Some of the darker neurons display a hypertrophy of the endoplasmic reticulum or a relative increase of neurofilaments. The mitochondria in some of these cells show early stages of degeneration. These changes appear 13 days after TOCP ingestion.     

Environmental history and physiological state influence feeding responses of Atrina zelandica to suspended sediment concentrations

Epifaunal suspension-feeding bivalves can play important roles in marine ecosystems affecting macrobenthic communities, benthic boundary layers and benthic-pelagic coupling, not just by their presence but also by any changes in feeding behaviour. While seston quality and quantity have consistently been found to be important influences on the feeding rates of suspension-feeding bivalves, factors stressing individuals are also likely to be important, as they may affect energy-dependent thresholds of response. We postulated that (1) history of seston quantity would affect how suspension feeders deal with increases in total suspended particulates, and (2) high-seston concentrations would affect feeding rates more in individuals whose energy reserves were low after spawning. Three sites were selected for short-term (1 day) feeding experiments on the pinnid bivalve, Atrina zelandica. At one site, the experiment was run pre-and postspawning. Atrina exhibited high rejection of filtered particles (mostly 75% to 100%) and high organic absorption efficiencies (0.9-1) at all seston levels. Strong differences in the response of feeding behaviour to increased seston concentrations were observed between A. zelandica from the different sites, with lesser differences observed between times. The site-specific feeding responses to seston concentrations observed are likely to affect our ability to model responses of A. zelandica to sediment loading and to influence the importance of A. zelandica to benthic-pelagic coupling.     

Control of Redox Balance by the Stringent Response Regulatory Protein Promotes Antioxidant Defenses of Salmonella

We report herein a critical role for the stringent response regulatory DnaK suppressor protein (DksA) in the coordination of antioxidant defenses. DksA helps fine-tune the expression of glutathione biosynthetic genes and discrete steps in the pentose phosphate pathway and tricarboxylic acid cycle that are associated with the generation of reducing power. Control of NAD(P)H/NAD(P)⁺ redox balance by DksA fuels downstream antioxidant enzymatic systems in nutritionally starving Salmonella. Conditional expression of the glucose-6-phosphate dehydrogenase-encoding gene zwf, shown here to be under DksA control, increases both the NADPH pool and antioxidant defenses of dksA mutant Salmonella. The DksA-mediated coordination of redox balance boosts the antioxidant defenses of stationary phase bacteria. Not only does DksA increase resistance of Salmonella against hydrogen peroxide (H₂O₂), but it also promotes fitness of this intracellular pathogen when exposed to oxyradicals produced by the NADPH phagocyte oxidase in an acute model of infection. Given the role of DksA in the adjustment of gene expression in most bacteria undergoing nutritional deprivation, our findings raise the possibility that the control of central metabolic pathways by this regulatory protein maintains redox homeostasis essential for antioxidant defenses in phylogenetically diverse bacterial species.     

Recent developments in the cell biology and biochemistry of glycosylphosphatidylinositol lipids (Review)

Glycosylphosphatidylinositols (GPIs) represent an abundant and ubiquitous class of eukaryotic glycolipids. Although these structures were originally discovered in the form of GPI-anchored cell surface glycoproteins, it is becoming increasingly clear that a significant proportion of the GPI synthetic output of a cell is not directed to protein anchoring. Indeed, pools of nonprotein-linked GPIs can approach 107 molecules per cell in some cell types, especially the protozoa, with a large proportion of these molecules being displayed at the cell surface. Recent studies which form the subject of this review indicate that there is (a) considerable diversity in the range of structural modifications found on GPI glycolipids within and between species and cell types, (b) complexity in the topological arrangement of the GPI biosynthetic pathway in the endoplasmic reticulum, and (c) spatial restriction of the biosynthetic pathway within the endoplasmic reticulum. Furthermore, consistent with additional functional roles for these lipids beyond serving as protein anchor precursors, products of the GPI biosynthetic pathway appear to be widely distributed in the cellular endomembrane system. These studies indicate that there is still much to learn about the organization of glycolipid biosynthetic pathways in eukaryotic cells, the nature and subcellular distribution of the lipid products of these pathways, and the function of these lipids within cells.     

Isopycnic-zonal centrifugation of plasma membrane,sarcoplasmic reticular fragments,lysosomes, and cytoplasmic proteins from phasic skeletal muscle

Homogenates of the posterior latissimus dorsi muscle, a phasic muscle, were fractionated by a one-step zonal centrifugation technique into four major organelle populations and cytoplasmic constituents. These were: (1) Plasma membrane fragments with a modal equilibrium density of 1.10 and containing 5′-nucleotidase, alkaline phosphodiesterase, $\text{p-}\text{nitrophenylphosphatase}$ and acid phosphatase (β-glycerophosphate was used as the substrate). (2) Sarcoplasmic reticular fragments which could be further subdivided into calcium transport vesicles, with a modal equilibrium density of 1.16, that exhibited calcium uptake; K⁺-ATPase; leucyl-β-naphthylamidase; acid phosphodiesterase; acid phosphatase (using cytidine monophosphate as the substrate); and sarcoplasmic reticular lysosomes, with a modal equilibrium density of 1.18, possessing dipeptidyl-aminopeptidase II, cathepsin D, α-glucosidase, $\text{N-}\text{acetyl}\text{-β-}\text{glucosaminidase}$, and NADH oxidase activity. (3) Mitochondria with a modal equilibrium density of 1.21. (4) Catalase-containing vesicles with a modal equilibrium density of 1.22; and cytoplasmic constituents (modal density of 1.25) with phosphorylase, pyruvate kinase, myosin-ATPase, aldolase, and protein and RNA content. The purity of these organelles was equal to or better than previous efforts, with a 30-fold purification achieved for 5′-nucleotidase and alkaline phosphodiesterase. These results lend support to the hypothesis that the sarcoplasmic reticulum of phasic muscle, in addition to its specialized role in excitation-contraction coupling, represents a multifunctional membrane system, and that, similar to the smooth endoplasmic reticulum of other cells, it includes some membrane-bound lysosomal enzymes and NADH oxidase.